WO2018057967A2

WO2018057967A2 - Constructs targeting hiv peptide/mhc complexes and uses thereof

Info

Publication number: WO2018057967A2
Application number: PCT/US2017/053073
Authority: WO
Inventors: Hong Liu; Vivien Wai-Fan CHAN; Lianxing Liu; Javier Morales; Zhiyuan Yang; Cheng Liu
Original assignee: Eureka Therapeutics, Inc.
Priority date: 2016-09-23
Filing date: 2017-09-22
Publication date: 2018-03-29
Also published as: WO2018057967A3; TW201827453A

Abstract

The present application provides constructs comprising an antibody moiety that specifically bind to a complex comprising a human immunodeficiency virus 1 (HIV-1) reverse transcriptase (RT) peptide and an MHC class I protein. Also provided are methods of making and using these constructs.

Description

CONSTRUCTS TARGETING HIV PEPTIDE/MHC COMPLEXES AND USES

THEREOF

CROSS-REREFENCE TO RELATED APPLICATION

[0001] This application claims priority to U.S. Provisional Application No. 62/399,210, filed on September 23, 2016, which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0002] This invention pertains to constructs comprising an antibody moiety that specifically bind to a complex comprising a human immunodeficiency virus 1 (HIV-1) reverse transcriptase (RT) peptide and an MHC class I protein, including methods of manufacture and uses thereof such as for treating and/or diagnosing diseases.

SUBMISSION OF SEQUENCE LISTING ON ASCII TEXT FILE

[0003] The content of the following submission on ASCII text file is incorporated herein by reference in its entirety: a computer readable form (CRF) of the Sequence Listing (file name: 750042000740SEQLIST.txt, date recorded: September 22, 2017, size: 145 KB).

BACKGROUND OF THE INVENTION

[0004] The Human Immunodeficiency Virus (HIV) is a lentivirus that causes HIV infection and acquired immunodeficiency syndrome (AIDS). Lentivirus is a genus of viruses of the

Retroviridae family. Lentiviruses are transmitted as single- stranded, positive- sense, enveloped RNA virus. HIV has been divided into two types: HIV-1 and HIV-2. HIV-1 is more virulent and more infective than HIV-2. HIV-1 is the cause of majority of HIV infections globally. HIV-2 has relatively poor capacity for transmission, it is found mainly in West Africa, with some cases in India and Europe.

[0005] HIV has several major genes coding for structural proteins as well as several accessory genes. The HIV genome contains three major genes, gag, pol, and env, encoding major structural proteins as well as essential viral enzymes. Pol codes reverse transcriptase (RT), RNase

H, integrase, and HIV protease. Reverse transcriptase is required to transcribe DNA from RNA template. Integrase is necessary to integrate the double- stranded viral DNA into the host genome. HIV protease is required to cleave the precursor Gag polyprotein to produce structural proteins.

[0006] Reverse transcriptase (RT) is a critical enzyme in the HIV replication cycle. RT is used by retroviruses to transcribe their single- stranded RNA genome into single- stranded DNA and to subsequently construct a complementary strand of DNA, providing a DNA double helix capable of integration into host cell chromosomes. Functional HIV1-RT is a heterodimer containing subunits of 66 kDa (p66) and 51 kDa (p51). p66 contains two domains, the N-terminal polymerase domain (440 residues) and the C-terminal RNase H domain (120 residues). p51 is processed by proteolytic cleavage of p66 and corresponds to the polymerase domain of the p66 subunit.

[0007] In chronically infected patients that do not start antiretroviral therapy (ART) early, up to 98% or more of latent virus may carry escape muatations that confer resistance to cytotoxic T lymphocytes (CTLs) directed at common epitopes, posing a significant hurdle in purging the latent reservoir of virus in these patients to effect a cure of the HIV infection (Deng, K. et al. (2015) Nature 517(7534): 381-385). For a review of various T cell gene-engineering and gene- editing strategies used in attempts to inhibit HIV-1 replication see Leibman, R. S., & Riley, J. L. (2015) Molecular Therapy 23(1): 1149-1159 and June, C. H., & Levine, B. L. (2015) Phil. Trans. R. Soc. B 370(1680): 20140374.

[0008] Recent advances in using phage display to generate mAbs have made it possible to select agents with exquisite specificity against defined epitopes from large antibody repertoires. A number of such mAbs specific for solid tumor antigens, in the context of HLA-A01 and HLA- A02, have been successfully selected from phage display libraries (Noy et al., Expert Rev.

Anticancer Ther. 5(3):523-536, 2005; Chames et al, Proc. Natl. Acad. Sci. USA 97:7969-79'/ '4, 2000; Held et al., Eur. J. Immunol. 34:2919-2929, 2004; Lev et al., Cancer Res. 62:3184-3194, 2002; Klechevsky et al, Cancer Res. 68(15):6360-6367, 2008). More recently, a human mAb specific for human WT1/HLA-A02 complex, a well-described T cell epitope, has been shown to inhibit multiple cancer cell lines and primary cancer cells via Fc-mediated effector cell function (Dao et al, Sci. Transl. Med. 5: 176ra33, 2013; Veomett et al, Clin. Cancer Res.

doi: 10.1158/1078-0432, 2014) in cellular assays and in in vivo models.

[0009] The disclosures of all publications, patents, patent applications and published patent applications referred to herein are hereby incorporated herein by reference in their entirety.

BRIEF SUMMARY OF THE INVENTION

[0010] The present application in one aspect provides constructs (such as isolated constructs) that bind to a complex comprising an HIV-1 RT peptide and an MHC class I protein (referred to herein as an "HIV-1 RT/MHC class I complex," or "RTMC"). In some embodiments, the constructs ("anti-RTMC constructs") comprise an antibody moiety (referred to herein as an "anti-RTMC antibody moiety") that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein.

[0011] Thus, in some embodiments, there is provided an anti-RTMC construct (such as an isolated anti-RTMC construct) comprising an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein. In some embodiments, the HIV-1 RT/MHC class I complex is present on a cell surface. In some embodiments, the HIV-1 RT/MHC class I complex is present on the surface of an immune cell, such as a T cell.

[0012] In some embodiments, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the MHC class I protein is HLA-A. In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is the HLA-A*02:01 subtype of the HLA-A02 allele.

[0013] In some embodiments, according to any of the anti-RTMC constructs (such as isolated anti-RTMC constructs) described above, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the antibody moiety cross-reacts with a complex comprising the HIV-1 RT peptide and a second MHC class I protein having a different HLA allele than the MHC class I protein. In some embodiments, the antibody moiety cross-reacts with a complex comprising a variant of the HIV-1 RT peptide comprising one amino acid substitution (such as a conservative amino acid substitution) and the MHC class I protein.

[0014] In some embodiments, according to any of the anti-RTMC constructs (such as isolated anti-RTMC construct) described above, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the HIV-1 RT peptide is about 8 to about 12 (such as about any of 8, 9, 10, 11, or 12) amino acids in length. In some embodiments, the HIV-1 RT peptide is derived from the region corresponding to amino acids 181-189 of SEQ ID NO: 1. In some embodiments, the HIV-

1 RT peptide comprises an amino acid sequence selected from the group consisting of SEQ ID

NOs: 5-18. In some embodiments, the HIV-1 RT peptide has the amino acid sequence

YQYVDDLYV (SEQ ID NO: 6). In some embodiments, the isolated anti-RTMC construct cross-reacts with a complex comprising a variant of the HIV-1 RT peptide having the amino acid sequence of any one of YQYMDDLYV (SEQ ID NO: 5), YQYIDDLYV (SEQ ID NO: 7),

CQYMDDLYV (SEQ ID NO: 8), or CQYVDDLYV (SEQ ID NO: 9) and the MHC class I protein. In some embodiments, the HIV-1 RT peptide has the amino acid sequence YQYMDDLYV (SEQ ID NO: 5). In some embodiments, the isolated anti-RTMC construct cross-reacts with a complex comprising a variant of the HIV-1 RT peptide having the amino acid sequence of any one of YQYVDDLYV (SEQ ID NO: 6), YQYIDDLYV (SEQ ID NO: 7), CQYMDDLYV (SEQ ID NO: 8), or CQYVDDLYV (SEQ ID NO: 9) and the MHC class I protein.

[0015] In some embodiments, according to any of the anti-RTMC constructs (such as isolated anti-RTMC constructs) described above, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the antibody moiety is a full-length antibody, a Fab, a Fab', a (Fab')2, an Fv, or a single chain Fv (scFv). In some embodiments, the antibody moiety is fully human, semisynthetic with human antibody framework regions, or humanized.

[0016] In some embodiments, according to any of the anti-RTMC constructs (such as isolated anti-RTMC constructs) described above, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the antibody moiety binds to the HIV-1 RT/MHC class I complex with an equilibrium dissociation constant (K_d) between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the isolated anti-RTMC construct binds to the HIV-1 RT/MHC class I complex with a IQ between about 0.1 pM to about

500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM,

50 nM, 100 nM, or 500 nM, including any ranges between these values).

[0017] In some embodiments, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the antibody moiety comprises: i) a heavy chain variable domain comprising a heavy chain complementarity determining region (HC-CDR) 1 comprising the amino acid sequence of

SEQ ID NO: 240, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ

ID NOs: 241-244, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of

SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising a light chain complementarity determining region (LC-CDR) 1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions.

[0018] In some embodiments, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the antibody moiety comprises: i) a heavy chain variable domain comprising an HC- CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC- CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions.

[0019] In some embodiments, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the antibody moiety comprises: i) a heavy chain variable domain comprising an HC-

CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96, an HC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 97-124, and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR regions; and ii) a light chain variable domain comprising an LC-

CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189, an LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207, and an LC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR regions.

[0020] In some embodiments, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the antibody moiety comprises a) a heavy chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 19-46 or a variant thereof having at least about 95% (such as at least about any of 95%, 96%, 97%, 98%, or 99%) sequence identify to any one of SEQ ID NOs: 19-46; and b) a light chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 47-74 or a variant thereof having at least about 95% (such as at least about any of 95%, 96%, 97%, 98%, or 99%) sequence identity to any one of SEQ ID NOs: 47-74. In some embodiments, the antibody moiety comprises a heavy chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 47-74.

[0021] In some embodiments, the anti-RTMC construct comprises a first antibody moiety that competes for binding to a target HIV-1 RT/MHC class I complex with a second antibody moiety according to any of the antibody moieties described above. In some embodiments, the first antibody moiety binds to the same, or substantially the same, epitope as the second antibody moiety. In some embodiments, binding of the first antibody moiety to the target HIV-1 RT/MHC class I complex inhibits binding of the second antibody moiety to the target HIV-1 RT/MHC class I complex by at least about 70% (such as by at least about any of 75%, 80%, 85%, 90%, 95%, 98% or 99%), or vice versa. In some embodiments, the first antibody moiety and the second antibody moiety cross-compete for binding to the target HIV-1 RT/MHC class I complex, i.e., each of the first and second antibody moieties competes with the other for binding to the target HIV-1 RT/MHC class I complex.

[0022] In some embodiments, according to any of the anti-RTMC constructs described above

(such as isolated anti-RTMC constructs), the isolated anti-RTMC construct is a full-length antibody. In some embodiments, the isolated anti-RTMC construct is monospecific. In some embodiments, the isolated anti-RTMC construct is multi- specific. In some embodiments, the isolated anti-RTMC construct is bispecific. In some embodiments, the isolated anti-RTMC molecule is a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into-hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a

heteromultimeric antibody, or a heteroconjugate antibody. In some embodiments, the isolated anti-RTMC construct is a tandem scFv comprising two scFvs linked by a peptide linker. In some embodiments, the peptide linker comprises (and in some embodiments consists of) the amino acid sequence of SEQ ID NO: 276.

[0023] In some embodiments, according to any of the anti-RTMC constructs described above (such as isolated anti-RTMC constructs), the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the isolated anti-RTMC construct further comprises a second antigen-binding moiety that specifically binds to a second antigen. In some embodiments, the second antigen- binding moiety is an antibody moiety. In some embodiments, the second antigen is an antigen on the surface of a T cell. In some embodiments, the T cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, and a natural killer T cell. In some embodiments, the second antigen is selected from the group consisting of CD3y, CD35, CD3s, CD3C, CD28, OX40, GITR, CD137, CD27, CD40L, and HVEM. In some embodiments, the second antigen is CD3s, and the isolated anti-RTMC construct is a tandem scFv comprising an N-terminal scFv specific for the HIV-1 RT/MHC class I complex and a C-terminal scFv specific for CD3s. In some embodiments, the second antigen is an antigen on the surface of a natural killer cell, a neutrophil, a monocyte, a macrophage, or a dendritic cell.

[0024] In some embodiments, according to any of the anti-RTMC constructs (such as isolated anti-RTMC constructs) described above, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the isolated anti-RTMC construct is a chimeric antigen receptor (CAR). In some embodiments, the chimeric antigen receptor comprises an extracellular domain comprising the antibody moiety, a transmembrane domain, and an intracellular signaling domain. In some embodiments, the intracellular signaling domain comprises a CD3ζ intracellular signaling sequence and a co-stimulatory signaling sequence. In some embodiments, the co-stimulatory signaling sequence is a CD28 or 4- IBB intracellular signaling sequence. In some embodiments, the the intracellular signaling domain comprises a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. [0025] In some embodiments, according to any of the anti-RTMC constructs (such as isolated anti-RTMC constructs) described above, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, wherein the isolated anti-RTMC construct is a a chimeric antibody/T cell receptor (abTCR). In some embodiments, the anti-RTMC abTCR comprises an extracellular domain comprising the antibody moiety and a T cell receptor (TCR) module (TCRM) comprising TCR transmembrane domains. In some embodiments, the TCRM is capable of recruiting at least one TCR-associated signaling module. In some embodiments, the TCR-associated signaling module is selected from the group consisting of CD35s, CD3y8, and ζζ. In some embodiments, the antibody moiety comprises: a) a first polypeptide chain comprising a first antigen-binding domain comprising V_H and C_HI antibody domains; and b) a second polypeptide chain comprising a second antigen-binding domain comprising V_L and C_L antibody domains, wherein the V_H and C_HI domains of the first antigen-binding domain and the V_L and C_L domains of the second antigen-binding domain form a Fab-like antigen-binding module that specifically binds to the RTMC.

[0026] In some embodiments, according to any of the anti-RTMC constructs (such as isolated anti-RTMC constructs) described above, the anti-RTMC construct comprises an antibody moiety that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, wherein the isolated anti-RTMC construct is an immunoconjugate comprising the antibody moiety and an effector molecule. In some embodiments, the effector molecule is a therapeutic agent selected from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid. In some embodiments, the therapeutic agent is a drug or a toxin. In some embodiments, the effector molecule is a label.

[0027] In yet other embodiments, there is provided a host cell expressing or associated with an anti-RTMC construct, or polypeptide component thereof. In some embodiments, there is provided a nucleic acid encoding an anti-RTMC construct, or polypeptide component thereof. In some embodiments, there is provided a vector comprising the nucleic acid. In some

embodiments, there is provided an effector cell expressing or associated with an anti-RTMC construct. In some embodiments, the effector cell is a T cell.

[0028] In yet other embodiments, there is provided a pharmaceutical composition comprising an anti-RTMC construct (such as an isolated anti-RTMC construct), a host cell, a nucleic acid, a vector, or an effector cell according to any of the embodiments described above. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0029] In some embodiments, there is provided a method of detecting a cell presenting a complex comprising an HIV-1 RT peptide and an MHC class I protein on its surface, comprising contacting the cell with an anti-RTMC construct (such as an isolated anti-RTMC construct) according to any of the embodiments described above comprising a) an antibody moiety that specifically binds to a complex comprising the HIV-1 RT peptide and the MHC class I protein and b) a label, and detecting the presence of the label on the cell.

[0030] In some embodiments, there is provided a method of treating an individual having an HIV- 1 infection, comprising administering to the individual an effective amount of a

pharmaceutical composition comprising an anti-RTMC construct (such as an isolated anti- RTMC construct) according to any of the embodiments described above. In some embodiments, the pharmaceutical composition further comprises a cell (such as an effector cell) associated with the isolated anti-RTMC construct. In some embodiments, there is provided a method of treating an individual having an HIV- 1 infection, comprising administering to the individual an effective amount of an effector cell expressing any of the anti-RTMC CARs or anti-RTMC abTCRs described above. In some embodiments, the effector cell is a T cell. In some

embodiments, the individual is human.

[0031] In some embodiments, there is provided a method of diagnosing an individual having an HIV-1 infection, comprising: a) administering an effective amount of an isolated anti-RTMC construct comprising a label according to any of the embodiments described above to the individual; and b) determining the level of the label in the individual, wherein a level of the label above a threshold level indicates that the individual has the HIV-1 infection. In some

embodiments, there is provided a method of diagnosing an individual having an HIV-1 infection, comprising: a) contacting a sample derived from the individual with an isolated anti-RTMC construct comprising a label according to any of the embodiments described above; and b) determining the number of cells bound with the isolated anti-RTMC construct in the sample, wherein a value for the number of cells bound with the isolated anti-RTMC construct above a threshold level indicates that the individual has the HIV-1 infection. In some embodiments, the individual is human.

[0032] Also provided are methods of making any of the constructs described herein, articles of manufacture, and kits that are suitable for the methods described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] FIG. 1 shows the results of BB7.2 FACS binding assays for T2 cells loaded with HIV-1 RT 181 peptides (WT, M184V, Ml 841, Y181C, or Y181C/M184V) or HIV-1 RT 181 peptides having single alanine substitutions at positions 1, 2, 3, 4, 5, 6, 7, 8, or 9. Unloaded cells only (T2 Cell), 2° antibody BB7.2 only (2^nd Antibody), and unloaded cells with 2° antibody BB7.2 (T Cell + 2^nd Antibody) conditions were included as controls.

[0034] FIG. 2 shows the T-cell killing of HIV-1 RT 181 M184V-loaded T2 cells mediated by anti-HIV-1 RT 181/HLA-A*02:01 bispecific antibodies prepared from various phage clones. Negative controls included T2 cells loaded with P20 control peptide mix.

[0035] FIG. 3 shows the T-cell killing of parental SK-Hepl cells and SK-Hepl cells transduced to express HIV-1 RT 181 WT, HIV-1 RT 181 Ml 84V, or HIV-1 RT 181 Ml 841 mediated by anti-HIV-1 RT 181/MHC bispecific antibodies (BsAb).

[0036] FIG. 4 shows flow cytometry analysis of T cells transduced with anti-HIV CAR 10, anti- HIV CAR 14, or mock-transfected, and stained with HIV-1 RT 181-189 peptide /HLA-A*02:01 tetramers.

[0037] FIG. 5 shows the killing of SK-HEP-1 and SK-HEP-l-MG cell lines mediated by T cells expressing anti-HIV CAR 10 or anti-HIV CAR 14.

DETAILED DESCRIPTION OF THE INVENTION

[0038] The present application provides isolated constructs (referred to herein as "anti-RTMC constructs") that comprise an antibody moiety (referred to herein as an "anti-RTMC antibody moiety") that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein (referred to herein as an "HIV-1 RT/MHC class I complex," or "RTMC").

[0039] Proteins expressed by viruses such as HIV represent excellent targets for immunotherapy due to absence of expression in normal cells. RT is an intracellular protein that has not been successfully targeted by cytotoxic effector cell therapies against cell surface proteins.

YQYMDDLYV is an HLA-A2 restricted HIV-1 epitope in the HIV-1 RT. The YQYMDDLYV epitope is one of the 14 most conserved regions of the HIV proteome in four major HIV-1 clades A, B, C and D (Letourneau, S., et al. (2007) PloS one, 2(10): e984; Harrer, E. et al. (1996) Journal of Infectious Diseases 173(2): 476-479). This epitope is of great clinical relevance because it lies within the active site of RT and is a target of many reverse transcriptase inhibitors. One of the most common mutations in RT that leads to drug resistance, M184V, lies in this region (Shafer, R. W., & Schapiro, J. M. (2008) AIDS reviews 10(2): 67).

[0040] The anti-RTMC constructs specifically recognize HIV- 1 RT/MHC class I complexes. In some embodiments, the HIV- 1 RT/MHC class I complex is on the surface of cells expressing HIV- 1 RT. Anti-RTMC constructs may specifically bind to the N-terminal portion, the C- terminal portion, or the middle portion of the HIV- 1 RT peptide in the complex, and/or cross- react with at least one complex comprising the HIV- 1 RT peptide and a different subtype of the MHC class I protein (e.g. , the anti-RTMC construct binds to both an HIV- 1 RT peptide/HLA- A*02:01 complex and an HIV- 1 RT peptide/HLA-A*02:02 complex). The anti-RTMC constructs allow for specific targeting of RTMC-presenting cells (i.e. , cells presenting on their surface an HIV- 1 RT peptide bound to an MHC molecule), such as infected cells expressing HIV- 1 RT. This strategy provides a significant technical advantage over using antibodies directed against the HIV- 1 RT protein, which may not specifically target RTMC-presenting cells. Furthermore, when fused to a detectable moiety, the anti-RTMC antibody moiety allows for diagnosis and prognosis of HIV- 1 infections with high sensitivity to changes in the number and distribution of RTMC-presenting cells.

[0041] Using phage display technology, we generated multiple monoclonal antigen-binding antibody fragments that are specific and high affinity against HIV- 1 RT 181 peptide/HLA- A*02:01 complex. Flow cytometry and T-cell mediated cytotoxicity assays demonstrated that the antibodies recognized HIV- 1 RT peptide-pulsed T2 cells in an HIV- 1 RT- and HLA- A*02:01 -restricted manner. When armed as anti-CD3 bispecific antibodies, the antibodies redirected human T cells to kill HIV- 1 RT-positive and HLA-A*02:01-positive target cells. The data presented herein demonstrate that antibodies against HIV- 1 RT peptides in the context of an HLA complex can be effective therapeutic agents for HIV- 1 infection.

[0042] The present application thus provides constructs (such as isolated constructs) comprising an antibody moiety that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein. The construct can be, for example, a full-length anti-RTMC antibody, a multi- specific anti-RTMC molecule (such as a bispecific anti-RTMC antibody), an anti-RTMC chimeric antigen receptor ("CAR"), or an anti-RTMC immunoconjugate.

[0043] In another aspect, there are provided nucleic acids encoding the anti-RTMC constructs or the anti-RTMC antibody moiety portion of the constructs.

[0044] In another aspect, there are provided compositions comprising an anti-RTMC construct comprising an antibody moiety that specifically binds to a complex comprising an HIV- 1 RT- peptide and an MHC class I protein. The composition can be a pharmaceutical composition comprising an anti-RTMC construct or an effector cell expressing or associated with the anti- RTMC construct (for example a T cell expressing an anti-RTMC CAR or anti-RTMC abTCR).

[0045] Also provided are methods of making and using the anti-RTMC constructs (or cells expressing or associated with the anti-RTMC constructs) for treatment or diagnostic purposes, as well as kits and articles of manufacture useful for such methods.

Definitions

[0046] As used herein, "treatment" or "treating" is an approach for obtaining beneficial or desired results, including clinical results. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviating one or more symptoms resulting from the disease, diminishing the extent of the disease, stabilizing the disease (e.g. , preventing or delaying the worsening of the disease), preventing or delaying the spread of the disease, preventing or delaying the recurrence of the disease, delay or slowing the progression of the disease, ameliorating the disease state, providing a remission (partial or total) of the disease, decreasing the dose of one or more other medications required to treat the disease, delaying the progression of the disease, increasing or improving the quality of life, increasing weight gain, and/or prolonging survival. Also encompassed by "treatment" is a reduction of pathological consequence of a disease. The methods of the invention contemplate any one or more of these aspects of treatment.

[0047] The term "refractory" or "resistant" refers to a disease that has not responded to treatment.

[0048] "Activation", as used herein in relation to a cell expressing CD3, refers to the state of the cell that has been sufficiently stimulated to induce a detectable increase in downstream effector functions of the CD3 signaling pathway, including, without limitation, cellular proliferation and cytokine production.

[0049] The term "antibody moiety" includes full-length antibodies and antigen-binding fragments thereof. A full-length antibody comprises two heavy chains and two light chains. The variable regions of the light and heavy chains are responsible for antigen binding. The variables region in both chains generally contain three highly variable loops called the complementarity determining regions (CDRs) (light chain (LC) CDRs including LC-CDR1, LC-CDR2, and LC- CDR3, heavy chain (HC) CDRs including HC-CDR1, HC-CDR2, and HC-CDR3). CDR boundaries for the antibodies and antigen-binding fragments disclosed herein may be defined or identified by the conventions of Kabat, Chothia, or Al-Lazikani (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). The three CDRs of the heavy or light chains are interposed between flanking stretches known as framework regions (FRs), which are more highly conserved than the CDRs and form a scaffold to support the hypervariable loops. The constant regions of the heavy and light chains are not involved in antigen binding, but exhibit various effector functions. Antibodies are assigned to classes based on the amino acid sequence of the constant region of their heavy chain. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, which are characterized by the presence of α, δ, ε, γ, and μ heavy chains, respectively. Several of the major antibody classes are divided into subclasses such as lgGl (γΐ heavy chain), lgG2 (γ2 heavy chain), lgG3 (γ3 heavy chain), lgG4 (γ4 heavy chain), IgAl (al heavy chain), or lgA2 (a2 heavy chain).

[0050] The term "antigen-binding fragment" as used herein refers to an antibody fragment including, for example, a diabody, a Fab, a Fab', a F(ab')2, an Fv fragment, a disulfide stabilized Fv fragment (dsFv), a (dsFv)2, a bispecific dsFv (dsFv-dsFv'), a disulfide stabilized diabody (ds diabody), a single-chain antibody molecule (scFv), an scFv dimer (bivalent diabody), a multispecific antibody formed from a portion of an antibody comprising one or more CDRs, a camelized single domain antibody, a nanobody, a domain antibody, a bivalent domain antibody, or any other antibody fragment that binds to an antigen but does not comprise a complete antibody structure. An antigen-binding fragment is capable of binding to the same antigen to which the parent antibody or a parent antibody fragment (e.g. , a parent scFv) binds. In some embodiments, an antigen-binding fragment may comprise one or more CDRs from a particular human antibody grafted to a framework region from one or more different human antibodies.

[0051] The term "epitope" as used herein refers to the specific group of atoms or amino acids on an antigen to which an antibody or antibody moiety binds. Two antibodies or antibody moieties may bind the same epitope within an antigen if they exhibit competitive binding for the antigen.

[0052] A "Fab-like antigen-binding module" refers to an antibody moiety that comprises a first polypeptide chain and a second polypeptide chain, wherein the first and second polypeptide chains comprise a VL antibody domain, a CL antibody domain, a VH antibody domain, and a CHI antibody domain. The VL and CL antibody domains may be on one chain with the VH and CHI antibody domains on the other chain, or the VL and CHI antibody domains may be on one chain with the VH and CL antibody domains on the other chain. In some embodiments, the first and second polypeptide chains are linked, such as by a peptide linkage, or by another chemical linkage, such as a disulfide linkage. [0053] As used herein, a first antibody moiety "competes" for binding to a target RTMC with a second antibody moiety when the first antibody moiety inhibits target RTMC binding of the second antibody moiety by at least about 50% (such as at least about any of 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%) in the presence of an equimolar concentration of the first antibody moiety, or vice versa. A high throughput process for "binning" antibodies based upon their cross-competition is described in PCT Publication No. WO 03/48731.

[0054] As use herein, the term "specifically binds" or "is specific for" refers to measurable and reproducible interactions, such as binding between a target and an antibody or antibody moiety, that is determinative of the presence of the target in the presence of a heterogeneous population of molecules, including biological molecules. For example, an antibody or antibody moiety that specifically binds to a target (which can be an epitope) is an antibody or antibody moiety that binds this target with greater affinity, avidity, more readily, and/or with greater duration than its bindings to other targets. In some embodiments, an antibody or antibody moiety that specifically binds to an antigen reacts with one or more antigenic determinants of the antigen (for example an HIV-1 RT peptide/MHC class I protein complex) with a binding affinity that is at least about 10 times its binding affinity for other targets.

[0055] The term "T cell receptor," or "TCR," refers to a heterodimeric receptor composed of αβ or γδ chains that pair on the surface of a T cell. Each α, β, γ, and δ chain is composed of two Ig- like domains: a variable domain (V) that confers antigen recognition through the

complementarity determining regions (CDR), followed by a constant domain (C) that is anchored to cell membrane by a connecting peptide and a transmembrane (TM) region. The TM region associates with the invariant subunits of the CD3 signaling apparatus. Each of the V domains has three CDRs. These CDRs interact with a complex between an antigenic peptide bound to a protein encoded by the major histocompatibility complex (pMHC) (Davis and Bjorkman (1988) Nature, 334, 395-402; Davis et al. (1998) Annu Rev Immunol, 16, 523-544; Murphy (2012), xix, 868 p.).

[0056] The term "TCR-associated signaling molecule" refers to a molecule having a

cytoplasmic immunoreceptor tyro sine-based activation motif (IT AM) that is part of the TCR- CD3 complex. TCR-associated signaling molecules include CD3y8, CD35s, and ζζ, and are essential for the signaling capacity of the TCR.

[0057] The term "module" when referring to a portion of a protein is meant to include structurally and/or functionally related portions of one or more polypeptides which make up the protein. For example, a transmembrane module of a dimeric receptor may refer to the portions of each polypeptide chain of the receptor that span the membrane. A module may also refer to related portions of a single polypeptide chain. For example, a transmembrane module of a monomeric receptor may refer to portions of the single polypeptide chain of the receptor that span the membrane. A module may also include only a single portion of a polypeptide.

[0058] An "isolated" anti-RTMC construct as used herein refers to an anti-RTMC construct that (1) is not associated with proteins found in nature, (2) is free of other proteins from the same source, (3) is expressed by a cell from a different species, or, (4) does not occur in nature.

[0059] The term "isolated nucleic acid" as used herein is intended to mean a genomic nucleic acid, cDNA, or nucleic acid of synthetic origin or some combination thereof, which by virtue of its origin the "isolated nucleic acid" (1) is not associated with all or a portion of a polynucleotide in which the "isolated nucleic acid" is found in nature, (2) is operably linked to a polynucleotide which it is not linked to in nature, or (3) does not occur in nature as part of a larger sequence.

[0060] As used herein, the term "CDR" or "complementarity determining region" is intended to mean the non-contiguous antigen combining sites found within the variable region of both heavy and light chain polypeptides. These particular regions have been described by Kabat et al, J. Biol. Chem. 252:6609-6616 (1977); Kabat et al, U.S. Dept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991); by Chothia et al, J. Mol. Biol.

196:901-917 (1987); and MacCallum et al, J. Mol. Biol. 262:732-745 (1996), where the definitions include overlapping or subsets of amino acid residues when compared against each other. Nevertheless, application of either definition to refer to a CDR of an antibody or grafted antibodies or variants thereof is intended to be within the scope of the term as defined and used herein. The amino acid residues which encompass the CDRs as defined by each of the above cited references are set forth below in Table 1 as a comparison.

TABLE 1: CDR DEFINITIONS

Kabat¹ Chothia² MacCallum³

V_H CDRl 31-35 26-32 30-35

V_HCDR2 50-65 53-55 47-58 V_H CDR3 95-102 96-101 93-101 V_L CDRl 24-34 26-32 30-36 V_L CDR2 50-56 50-52 46-55 V_L CDR3 89-97 91-96 89-96

Residue numbering follows the nomenclature of Kabat et al., supra

2Residue numbering follows the nomenclature of Chothia et al., supra

3Residue numbering follows the nomenclature of MacCallum et al., supra [0061] The term "chimeric antibodies" refer to antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit a biological activity of this invention (see U.S. Patent No. 4,816,567; and Morrison et ah, Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).

[0062] The term "semi- synthetic" in reference to an antibody or antibody moiety means that the antibody or antibody moiety has one or more naturally occurring sequences and one or more non- naturally occurring (i.e., synthetic) sequences.

[0063] "Fv" is the minimum antibody fragment which contains a complete antigen-recognition and -binding site. This fragment consists of a dimer of one heavy- and one light-chain variable region domain in tight, non-covalent association. From the folding of these two domains emanate six hypervariable loops (3 loops each from the heavy and light chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0064] "Single-chain Fv," also abbreviated as "sFv" or "scFv," are antibody fragments that comprise the V_H and V_L antibody domains connected into a single polypeptide chain. In some embodiments, the scFv polypeptide further comprises a polypeptide linker between the V_H and V_L domains which enables the scFv to form the desired structure for antigen binding. For a review of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer- Verlag, New York, pp. 269-315 (1994).

[0065] The term "diabodies" refers to small antibody fragments prepared by constructing scFv fragments (see preceding paragraph) typically with short linkers (such as about 5 to about 10 residues) between the V_H and V_L domains such that inter-chain but not intra-chain pairing of the V domains is achieved, resulting in a bivalent fragment, i.e., fragment having two antigen- binding sites. Bispecific diabodies are heterodimers of two "crossover" scFv fragments in which the V_H and V_L domains of the two antibodies are present on different polypeptide chains. Diabodies are described more fully in, for example, EP 404,097; WO 93/11161; and Hollinger et al, Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993).

[0066] "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a

hypervariable region (HVR) of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or non-human primate having the desired antibody specificity, affinity, and capability. In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature 321:522- 525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).

[0067] "Percent (%) amino acid sequence identity" or "homology" with respect to the polypeptide and antibody sequences identified herein is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the

polypeptide being compared, after aligning the sequences considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full-length of the sequences being compared. For purposes herein, however, % amino acid sequence identity values are generated using the sequence comparison computer program MUSCLE (Edgar, R.C., Nucleic Acids Research 32(5): 1792- 1797, 2004; Edgar, R.C., BMC Bioinformatics 5(1): 113, 2004). [0068] The terms "Fc receptor" or "FcR" are used to describe a receptor that binds to the Fc region of an antibody. In some embodiments, an FcR of this invention is one that binds an IgG antibody (a γ receptor) and includes receptors of the FcyRI, FcyRII, and FcyRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FcyRII receptors include FcyRIIA (an "activating receptor") and FcyRIIB (an "inhibiting receptor"), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof.

Activating receptor FcyRIIA contains an immunoreceptor tyro sine-based activation motif (IT AM) in its cytoplasmic domain. Inhibiting receptor FcyRIIB contains an immunoreceptor tyro sine-based inhibition motif (ITIM) in its cytoplasmic domain (see review M. in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). The term includes allotypes, such as FcyRIIIA allotypes: FcYRIIIA-Phel58, FcYRIIIA-Vall58, FcyRIIA-R131 and/or FcyRIIA-Hm. FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al.,

Immunomethods 4:25-34 (1994); and de Haas et al, J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol . 24:249 (1994)).

[0069] The term "FcRn" refers to the neonatal Fc receptor (FcRn). FcRn is structurally similar to major histocompatibility complex (MHC) and consists of an oc-chain noncovalently bound to 2-microglobulin. The multiple functions of the neonatal Fc receptor FcRn are reviewed in Ghetie and Ward (2000) Annu. Rev. Immunol. 18, 739-766. FcRn plays a role in the passive delivery of immunoglobulin IgGs from mother to young and the regulation of serum IgG levels. FcRn can act as a salvage receptor, binding and transporting pinocytosed IgGs in intact form both within and across cells, and rescuing them from a default degradative pathway.

[0070] The "C_HI domain" of a human IgG Fc region (also referred to as "CI" of "HI" domain) usually extends from about amino acid 118 to about amino acid 215 (EU numbering system).

[0071] "Hinge region" is generally defined as stretching from Glu216 to Pro230 of human IgGl (Burton, Molec. Immunol.22: l6l-206 (1985)). Hinge regions of other IgG isotypes may be aligned with the IgGl sequence by placing the first and last cysteine residues forming inter- heavy chain S-S bonds in the same positions.

[0072] The "CH2 domain" of a human IgG Fc region (also referred to as "C2" of "H2" domain) usually extends from about amino acid 231 to about amino acid 340. The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for the domain-domain pairing and help stabilize the CH2 domain. Burton, Molec Immunol. 22: 161-206 (1985).

[0073] The "CH3 domain" (also referred to as "C2" or "H3" domain) comprises the stretch of residues C-terminal to a CH2 domain in an Fc region (i.e. from about amino acid residue 341 to the C-terminal end of an antibody sequence, typically at amino acid residue 446 or 447 of an

IgG).

[0074] A "functional Fc fragment" possesses an "effector function" of a native sequence Fc region. Exemplary "effector functions" include Clq binding; complement dependent

cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell- mediated cytotoxicity

(ADCC); phagocytosis; down regulation of cell surface receptors (e.g. B cell receptor; BCR), etc. Such effector functions generally require the Fc region to be combined with a binding domain (e.g. an antibody variable domain) and can be assessed using various assays known in the art.

[0075] An antibody with a variant IgG Fc with "altered" FcR binding affinity or ADCC activity is one which has either enhanced or diminished FcR binding activity (e.g. , FcyR or FcRn) and/or

ADCC activity compared to a parent polypeptide or to a polypeptide comprising a native sequence Fc region. The variant Fc which "exhibits increased binding" to an FcR binds at least one FcR with higher affinity (e.g. , lower apparent K_d or IC₅₀ value) than the parent polypeptide or a native sequence IgG Fc. According to some embodiments, the improvement in binding compared to a parent polypeptide is about 3 fold, such as about any of 5, 10, 25, 50, 60, 100,

150, 200, or up to 500 fold, or about 25% to 1000% improvement in binding. The polypeptide variant which "exhibits decreased binding" to an FcR, binds at least one FcR with lower affinity

(e.g., higher apparent IQ or higher IC₅₀ value) than a parent polypeptide. The decrease in binding compared to a parent polypeptide may be about 40% or more decrease in binding.

[0076] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted Ig bound to Fc receptors (FcRs) present on certain cytotoxic cells

(e.g. Natural Killer (NK) cells, neutrophils, and macrophages) enable these cytotoxic effector cells to bind specifically to an antigen-bearing target cell and subsequently kill the target cell with cytotoxins. The antibodies "arm" the cytotoxic cells and are absolutely required for such killing. The primary cells for mediating ADCC, NK cells, express FcyRIII only, whereas monocytes express FcyRI, FcyRII and FcyRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). To assess ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in US Patent No. 5,500,362 or 5,821,337 may be performed. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).

[0077] The polypeptide comprising a variant Fc region which "exhibits increased ADCC" or mediates antibody-dependent cell- mediated cytotoxicity (ADCC) in the presence of human effector cells more effectively than a polypeptide having wild type IgG Fc or a parent polypeptide is one which in vitro or in vivo is substantially more effective at mediating ADCC, when the amounts of polypeptide with variant Fc region and the polypeptide with wild type Fc region (or the parent polypeptide) in the assay are essentially the same. Generally, such variants will be identified using any in vitro ADCC assay known in the art, such as assays or methods for determining ADCC activity, e.g. in an animal model etc. In some embodiments, the variant is from about 5 fold to about 100 fold, e.g. from about 25 to about 50 fold, more effective at mediating ADCC than the wild type Fc (or parent polypeptide) .

[0078] "Complement dependent cytotoxicity" or "CDC" refers to the lysis of a target cell in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (Clq) to antibodies (of the appropriate subclass) which are bound to their cognate antigen. To assess complement activation, a CDC assay, e.g. as described in Gazzano-Santoro et al., J. Immunol. Methods 202: 163 (1996), may be performed. Polypeptide variants with altered Fc region amino acid sequences and increased or decreased Clq binding capability are described in US patent No. 6,194,551B 1 and

W099/51642. The contents of those patent publications are specifically incorporated herein by reference. See, also, Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[0079] Unless otherwise specified, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intra n(s).

[0080] The term "operably linked" refers to functional linkage between a regulatory sequence and a heterologous nucleic acid sequence resulting in expression of the latter. For example, a first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary to join two protein coding regions, in the same reading frame.

[0081] "Homologous" refers to the sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. When a position in both of the two compared sequences is occupied by the same base or amino acid monomer subunit, e.g., if a position in each of two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The percent of homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared times 100. For example, if 6 of 10 of the positions in two sequences are matched or homologous then the two sequences are 60% homologous. By way of example, the DNA sequences ATTGCC and TATGGC share 50% homology. Generally, a comparison is made when two sequences are aligned to give maximum homology.

[0082] An "effective amount" of an anti-RTMC construct or composition as disclosed herein, is an amount sufficient to carry out a specifically stated purpose. An "effective amount" can be determined empirically and by known methods relating to the stated purpose.

[0083] The term "therapeutically effective amount" refers to an amount of an anti-RTMC construct or composition as disclosed herein, effective to "treat" a disease in an individual. In the case of HIV- 1 infection, the therapeutically effective amount of the anti-RTMC construct or composition as disclosed herein can reduce the number of HIV- 1 infected cells; reduce HIV- 1 replication; inhibit (i.e., slow to some extent and preferably stop) spread of the infection to uninfected cells; and/or relieve to some extent one or more of the symptoms associated with the HIV- 1 infection. To the extent the anti-RTMC construct or composition as disclosed herein can kill existing HIV- 1 -infected cells, it can be cytotoxic. In some embodiments, the therapeutically effective amount is an amount that extends the survival of a patient.

[0084] As used herein, by "pharmaceutically acceptable" or "pharmacologically compatible" is meant a material that is not biologically or otherwise undesirable, e.g. , the material may be incorporated into a pharmaceutical composition administered to a patient without causing any significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained. Pharmaceutically acceptable carriers or excipients have preferably met the required standards of toxicological and manufacturing testing and/or are included on the Inactive Ingredient Guide prepared by the U.S. Food and Drug administration.

[0085] The term "label" when used herein refers to a detectable compound or composition which can be conjugated directly or indirectly to the anti-RTMC antibody moiety. The label may be detectable by itself (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable.

[0086] It is understood that embodiments of the invention described herein include "consisting" and/or "consisting essentially of embodiments.

[0087] Reference to "about" a value or parameter herein includes (and describes) variations that are directed to that value or parameter per se. For example, description referring to "about X" includes description of "X".

[0088] As used herein, reference to "not" a value or parameter generally means and describes "other than" a value or parameter. For example, the method is not used to treat infection of type X means the method is used to treat infections of types other than X.

[0089] As used herein and in the appended claims, the singular forms "a," "or," and "the" include plural referents unless the context clearly dictates otherwise.

Anti-RTMC constructs

[0090] In one aspect, the present invention provides HIV- 1 RT/MHC class I complex- specific constructs (anti-RTMC constructs) that comprise an antibody moiety that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein ("HIV- 1 RT/MHC class I complex," or "RTMC"). In some embodiments, the anti-RTMC construct is an isolated anti- RTMC construct. The specificity of the anti-RTMC construct derives from an anti-RTMC antibody moiety, such as a full-length antibody or antigen-binding fragment thereof, that specifically binds to the RTMC. In some embodiments, reference to a moiety (such as an antibody moiety) that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein means that the moiety binds to the RTMC with a) an affinity that is at least about 10 (including for example at least about any of 10, 20, 30, 40, 50, 75, 100, 200, 300, 400, 500, 750, 1000 or more) times its binding affinity for each of full-length HIV- 1 RT, free HIV- 1 RT peptide, MHC class I protein not bound to a peptide, and/or MHC class I protein bound to a non- HIV- 1 RT peptide; or b) a IQ no more than about 1/10 (such as no more than about any of 1/10, 1/20, 1/30, 1/40, 1/50, 1/75, 1/100, 1/200, 1/300, 1/400, 1/500, 1/750, 1/1000 or less) times its K_d for binding to each of full-length HIV-1 RT, free HIV-1 RT peptide, MHC class I protein not bound to a peptide, and/or MHC class I protein bound to a non- HIV-1 RT peptide. Binding affinity can be determined by methods known in the art, such as ELISA, fluorescence activated cell sorting (FACS) analysis, or radioimmunoprecipitation assay (RIA). IQ can be determined by methods known in the art, such as surface plasmon resonance (SPR) assay utilizing, for example, Biacore instruments, or kinetic exclusion assay (KinExA) utilizing, for example, Sapidyne instruments.

[0091] Contemplated anti-RTMC constructs include, for example, full-length anti-RTMC antibodies, multi- specific (such as bispecific) anti-RTMC molecules, anti-RTMC chimeric antigen receptors (CARs), and anti-RTMC immunoconjugates.

[0092] For example, in some embodiments, there is provided an anti-RTMC construct (such as an isolated anti-RTMC construct) comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein. In some embodiments, the HIV-1 RT peptide comprises (such as consists of) the amino acid sequence of any one of SEQ ID NOs: 5-18. In some embodiments, the HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01 (GenBank Accession No.: AAO20853). In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an

immunoconjugate. In some embodiments, the anti-RTMC construct binds the RTMC with a IQ between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein. [0093] In some embodiments, there is provided an anti-RTMC construct comprising an anti- RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide comprising (such as consisting of) the amino acid sequence of any one of SEQ ID NOs: 5-18 and HLA-A*02:01. In some embodiments, the HIV1- RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the anti-RTMC construct binds the RTMC with a IQ between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0094] In some embodiments, there is provided an anti-RTMC construct comprising an anti- RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247- 249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID

NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the anti- RTMC construct binds the RTMC with a IQ between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0095] In some embodiments, there is provided an anti-RTMC construct comprising an anti- RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising (and in some

embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an HC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 97-124; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain sequence comprising an LC- CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207; or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and an LC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, there is provided an anti- RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the anti-RTMC construct binds the RTMC with a IQ between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0096] In some embodiments, there is provided an anti-RTMC construct comprising an anti- RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises a heavy chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full- length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the anti-RTMC construct binds the RTMC with a K_d between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC construct cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0097] In some embodiments, there is provided an anti-RTMC construct comprising a first anti- RTMC antibody moiety that competes for binding to a target HIV-1 RT/MHC class I complex with a second anti-RTMC antibody moiety according to any of the anti-RTMC antibody moieties described herein. In some embodiments, the first anti-RTMC antibody moiety binds to the same, or substantially the same, epitope as the second anti-RTMC antibody moiety. In some embodiments, binding of the first anti-RTMC antibody moiety to the target HIV-1 RT/MHC class I complex inhibits binding of the second anti-RTMC antibody moiety to the target HIV-1 RT/MHC class I complex by at least about 70% (such as by at least about any of 75%, 80%, 85%, 90%, 95%, 98% or 99%), or vice versa. In some embodiments, the first anti-RTMC antibody moiety and the second anti-RTMC antibody moiety cross-compete for binding to the target HIV-1 RT/MHC class I complex, i.e., each of the first and second antibody moieties competes with the other for binding to the target HIV-1 RT/MHC class I complex.

[0098] For example, in some embodiments, there is provided an anti-RTMC construct comprising an anti-RTMC antibody moiety that competes for binding to a target HIV-1

RT/MHC class I complex with an antibody moiety comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions.

[0099] In some embodiments, there is provided an anti-RTMC construct comprising an anti-

RTMC antibody moiety that competes for binding to a target HIV-1 RT/MHC class I complex with an antibody moiety comprising i) a heavy chain variable domain sequence comprising an

HC-CDRl comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an HC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 97-124; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about

5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an

LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207; or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and an LC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, there is provided an anti-RTMC construct comprising an anti-RTMC antibody moiety that competes for binding to a target HIV-1 RT/MHC class I complex with an antibody moiety comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising (and in some

embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences.

[0100] In some embodiments, there is provided an anti-RTMC construct comprising an anti- RTMC antibody moiety that competes for binding to a target HIV-1 RT/MHC class I complex with an antibody moiety comprising a heavy chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising (and in some

embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, there is provided an anti-RTMC construct comprising an anti-RTMC antibody moiety that competes for binding to a target HIV-1

RT/MHC class I complex with an antibody moiety comprising a heavy chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 19-46, and a light chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 47-74.

[0101] In some embodiments, the anti-RTMC construct is stable in a solution for at least about 1 month (such as at least about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, or longer).

Stability can be expressed, for example, as the retention of an effector function (e.g., target cell- killing activity) of the anti-RTMC construct in an aqueous formulation kept at a storage temperature, e.g., 4° C. [0102] For example, in some embodiments, the anti-RTMC construct retains at least 40% (such as at least about any of 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or greater) of an effector function in an aqueous formulation kept at a storage temperature for at least about 1 month (such as at least about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, or longer). In some embodiments, the storage temperature is no greater than about 25° C (such as no greater than about any of 20, 18, 16, 14, 12, 10, 8, 6, 5, 4, 3, 2, 1, or 0° C, or less). In some embodiments, the effector function is target cell-killing activity.

[0103] In some embodiments, the antibody moiety of the anti-RTMC construct comprises HC- CDR and LC-CDR sequences, and a tandem di-scFv bispecific anti-RTMC antibody comprising the HC-CDR and LC-CDR sequences retains at least 40% (such as at least about any of 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or greater) of an effector function in an aqueous formulation kept at a storage temperature for at least about 1 month (such as at least about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, or longer), wherein the tandem di-scFv bispecific anti-RTMC antibody comprises a) a first scFv that specifically binds to a complex comprising an HIV-1 RT 181 peptide and an MHC class I protein and comprises the HC-CDR and LC-CDR sequences, and b) a second scFv that specifically binds to CD3s.. In some embodiments, the storage temperature is no greater than about 25° C (such as no greater than about any of 20, 18, 16, 14, 12, 10, 8, 6, 5, 4, 3, 2, 1, or 0° C, or less). In some embodiments, the effector function is target cell-killing activity.

[0104] The different aspects are discussed in various sections below in further detail.

Anti-RTMC antibody moiety

[0105] The anti-RTMC constructs comprise an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein.

[0106] In some embodiments, the anti-RTMC antibody moiety specifically binds to an RTMC present on the surface of a cell. In some embodiments, the cell is a T cell. In some embodiments, the T cell is a CD4⁺ T cell.

[0107] In some embodiments, the HIV-1 RT peptide is an MHC class I-restricted peptide. In some embodiments, the HIV-1 RT peptide is from about 8 to about 12 (such as about any of 8, 9, 10, 11, or 12) amino acids in length. [0108] In some embodiments, the HIV-1 RT peptide comprises (and in some embodiments consists of) the amino acid sequence of any one of SEQ ID NOs: 5-18.

[0109] In some embodiments, the HIV-1 RT peptide comprises (and in some embodiments consists of) the sequence of amino acids 181-189 of HIV-1 RT (YQYMDDLYV, SEQ ID NO: 5; also referred to herein as "HIV-1 RT 181").

[0110] In some embodiments, the MHC class I protein is HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, or HLA-G. In some embodiments, the MHC class I protein is HLA-A. In some embodiments, the HLA-A is HLA-A02. In some embodiments, the HLA-A02 is HLA-A*02:01.

[0111] In some embodiments, the anti-RTMC antibody moiety is a full-length antibody. In some embodiments, the anti-RTMC antibody moiety is an antigen-binding fragment, for example an antigen-binding fragment selected from the group consisting of a Fab, a Fab', a F(ab')2, an Fv fragment, a disulfide stabilized Fv fragment (dsFv), and a single-chain antibody molecule (scFv). In some embodiments, the anti-RTMC antibody moiety is an scFv. In some

embodiments, the anti-RTMC antibody moiety is human, humanized, or semi- synthetic.

[0112] In some embodiments, the anti-RTMC antibody moiety specifically binds to the N- terminal portion of the HIV-1 RT peptide in the complex. In some embodiments, the anti-RTMC antibody moiety specifically binds to the C-terminal portion of the HIV-1 RT peptide in the complex. In some embodiments, the anti-RTMC antibody moiety specifically binds to the middle portion of the HIV- 1 RT peptide in the complex.

[0113] In some embodiments, the anti-RTMC antibody moiety (or the anti-RTMC construct comprising the anti-RTMC antibody moiety) binds to the complex comprising the HIV-1 RT peptide and the MHC class I protein with an affinity that is at least about 10 (including for example at least about any of 10, 20, 30, 40, 50, 75, 100, 200, 300, 400, 500, 750, 1000 or more) times its binding affinity for each of full-length HIV-1 RT, free HIV-1 RT peptide, MHC class I protein not bound to a peptide, and/or MHC class I protein bound to a non- HIV-1 RT peptide. In some embodiments, the anti-RTMC antibody moiety (or the anti-RTMC construct comprising the anti-RTMC antibody moiety) binds to the complex comprising the HIV-1 RT peptide and the

MHC class I protein with a K_d no more than about 1/10 (such as no more than about any of 1/10,

1/20, 1/30, 1/40, 1/50, 1/75, 1/100, 1/200, 1/300, 1/400, 1/500, 1/750, 1/1000 or less) times its

K_d for binding to each of full-length HIV-1 RT, free HIV-1 RT peptide, MHC class I protein not bound to a peptide, and/or MHC class I protein bound to a non- HIV-1 RT peptide.

[0114] In some embodiments, the anti-RTMC antibody moiety (or the anti-RTMC construct comprising the anti-RTMC antibody moiety) binds to the complex comprising the HIV-1 RT peptide and the MHC class I protein with a K_d between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the anti-RTMC antibody moiety (or the anti-RTMC construct comprising the anti-RTMC antibody moiety) binds to the complex comprising the HIV-1 RT peptide and the MHC class I protein with a K_d between about 1 pM to about 250 pM (such as about any of 1, 10, 25, 50, 75, 100, 150, 200, or 250 pM, including any ranges between these values). In some embodiments, the anti-RTMC antibody moiety (or the anti-RTMC construct comprising the anti-RTMC antibody moiety) binds to the complex comprising the HIV-1 RT peptide and the MHC class I protein with a K_d between about 1 nM to about 500 nM (such as about any of 1, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 nM, including any ranges between these values).

[0115] In some embodiments, the anti-RTMC antibody moiety specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety cross-reacts with at least one complex comprising the HIV-1 RT peptide and an allelic variant of the MHC class I protein. In some embodiments, the allelic variant has up to about 10 (such as about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) amino acid substitutions when compared to the MHC class I protein. In some embodiments, the allelic variant is the same serotype as the MHC class I protein. In some embodiments, the allelic variant is a different serotype than the MHC class I protein. In some embodiments, the anti-RTMC antibody moiety does not cross- react with any complex comprising the HIV-1 RT peptide and an allelic variant of the MHC class I protein. In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0116] In some embodiments, the anti-RTMC antibody moiety specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety cross-reacts with at least one complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC antibody moiety does not cross-react with any complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide.

[0117] For example, in some embodiments, the anti-RTMC antibody moiety specifically binds to a complex comprising an HIV-1 RT peptide comprising (such as consisting of) the amino acid sequence of any one of SEQ ID NOs: 5-9 (SEQ ID NO: 5) and an MHC class I protein (such as

HLA-A02, for example HLA-A*02:01), wherein the anti-RTMC antibody moiety further binds to at least one (including at least about any of 2, 3, 4, or 5) of: a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 10 and an MHC class I protein (such as HLA- A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 11 and an MHC class I protein (such as HLA-A02, for example HLA- A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 12 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 13 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 15 and an MHC class I protein (such as HLA- A02, for example HLA-A*02:01).

[0118] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of any one of SEQ ID NOs: 5-9 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 10 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 11 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0119] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of any one of SEQ ID NOs: 6-9 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 10 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 12 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0120] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of any one of SEQ ID NOs: 5-9 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 10 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 11 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 13 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01). [0121] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of any one of SEQ ID NOs: 5-9 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 10 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 11 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 12 and an MHC class I protein (such as HLA-A02, for example HLA- A*02:01); and a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 13 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0122] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of SEQ ID NO: 6 or 7 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 11 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 12 and an MHC class I protein (such as HLA-A02, for example HLA- A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 13 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 15 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0123] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of any one of SEQ ID NOs: 5-9 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 10 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 11 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 12 and an MHC class I protein (such as HLA-A02, for example HLA- A*02:01); a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 13 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an alanine- substituted HIV-1 RT peptide of SEQ ID NO: 15 and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01). [0124] In some embodiments, the anti-RTMC antibody moiety specifically binds to a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of any one of SEQ ID NOs: 5-9 and HLA-A*02:01, wherein the anti-RTMC antibody moiety cross-reacts with at least one (including at least about any of 2, 3, 4, 5, or 6) of: a complex comprising the HIV-1 RT peptide and HLA-A*02:02 (GenBank Accession No.: AFL91480), a complex comprising the HIV-1 RT peptide and HLA-A*02:03 (GenBank Accession No.: AAA03604), a complex comprising the HIV-1 RT peptide and HLA-A*02:05 (GenBank Accession No.: AAA03603), a complex comprising the HIV-1 RT peptide and HLA-A*02:06 (GenBank Accession No.:

CCB78868), a complex comprising the HIV-1 RT peptide and HLA-A*02:07 (GenBank Accession No.: ACR55712), and a complex comprising the HIV-1 RT peptide and HLA- A*02: l l (GenBank Accession No.: CAB56609).

[0125] In some embodiments, the anti-RTMC antibody moiety specifically binds to one or more of: a complex comprising HIV-1 RT 181 (SEQ ID NO: 5) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYVDDLYV (SEQ ID NO: 6) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYIDDLYV (SEQ ID NO: 7) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of CQYMDDLYV (SEQ ID NO: 8) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of CQYVDDLYV (SEQ ID NO: 9) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0126] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide of SEQ ID NO: 5 and an MHC class I protein (such as HLA- A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYVDDLYV (SEQ ID NO: 6) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYIDDLYV (SEQ ID NO: 7) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0127] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT peptide of SEQ ID NO: 5 and an MHC class I protein (such as HLA-

A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYVDDLYV (SEQ ID NO: 6) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYIDDLYV (SEQ ID NO: 7) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of CQYMDDLYV (SEQ ID NO: 8) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of CQYVDDLYV (SEQ ID NO: 9) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0128] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYVDDLYV (SEQ ID NO: 6) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of

YQYIDDLYV (SEQ ID NO: 7) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of CQYMDDLYV (SEQ ID NO: 8) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of CQYVDDLYV (SEQ ID NO: 9) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0129] In some embodiments, the anti-RTMC antibody moiety specifically binds to: a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of YQYVDDLYV (SEQ ID NO: 6) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01); and a complex comprising an HIV-1 RT 181 variant having the amino acid sequence of

YQYIDDLYV (SEQ ID NO: 7) and an MHC class I protein (such as HLA-A02, for example HLA-A*02:01).

[0130] In some embodiments, the anti-RTMC antibody moiety is a semi- synthetic antibody moiety comprising fully human sequences and one or more synthetic regions. In some embodiments, the anti-RTMC antibody moiety is a semi- synthetic antibody moiety comprising a fully human light chain variable domain and a semi- synthetic heavy chain variable domain comprising fully human FR1, HC-CDR1, FR2, HC-CDR2, FR3, and FR4 regions and a synthetic HC-CDR3. In some embodiments, the semi- synthetic heavy chain variable domain comprises a fully synthetic HC-CDR3 having a sequence from about 5 to about 25 (such as about any of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) amino acids in length. In some embodiments, the semi- synthetic heavy chain variable domain or the synthetic HC-CDR3 is obtained from a semi- synthetic library (such as a semi- synthetic human library) comprising fully synthetic HC-CDR3s having a sequence from about 5 to about 25 (such as about any of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) amino acids in length, wherein each amino acid in the sequence is randomly selected from the standard human amino acids, minus cysteine. In some embodiments, the synthetic HC-CDR3 is from about 7 to about 15 (such as about any of 7, 8, 9, 10, 11, 12, 13, 14, or 15) amino acids in length.

[0131] The anti-RTMC antibody moieties in some embodiments comprise specific sequences or certain variants of such sequences. In some embodiments, the amino acid substitutions in the variant sequences do not substantially reduce the ability of the anti-RTMC antibody moiety to bind the RTMC. For example, alterations that do not substantially reduce RTMC binding affinity may be made. Alterations that substantially improve RTMC binding affinity or affect some other property, such as specificity and/or cross-reactivity with related variants of the RTMC, are also contemplated.

[0132] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR3 comprising the amino acid sequence of any one of

SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of

1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-

CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions.

[0133] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR3 comprising the amino acid sequence of any one of

SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253.

[0134] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO:

240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID

NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or

3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of

CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions.

[0135] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253.

[0136] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC- CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253; or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions in the LC-CDR sequences.

[0137] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253. The sequences of the CDRs noted herein are provided in Table 2 below.

TABLE 2

consensus 4 NO: 253 X₄₄ = any AA

[0138] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC- CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions.

[0139] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain comprising an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239. [0140] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of 1, 2,

3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions.

[0141] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3,

4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid

substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239.

[0142] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of

SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ

ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences.

[0143] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, wherein the amino acid substitutions are in HC-CDRl or HC- CDR2; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, wherein the amino acid substitutions are in HC-CDRl or HC-CDR2.

[0144] In some embodiments, the anti-RTMC antibody moiety comprises i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239. The sequences of the HC-CDRs from putative anti-RTMC antibody clones are provided in Table 3 below and the LC- CDRs from the clones are provided in Table 4 below.

TABLE 3

GFTFGDYG INWNGGST ARDPIYSGSYKAFDY SEQID NO: 75 SEQID NO: 97 SEQID NO: 125

GYSFTSYW IDPSDSYT ARYDFWSGSDY SEQID NO: 76 SEQID NO: 98 SEQID NO: 126

GFTFSNYA ISRTGATI VKDLFGADSGYDGVAYYFGD SEQID NO: 77 SEQID NO: 99 SEQID NO: 127

GYYFSAYY INPSGGST ARGSDYYPFNHWGDL SEQID NO: 78 SEQID NO: 100 SEQID NO: 128

GFTFSDYY ITGKSSYT ARYPSYVEGDY SEQID NO: 79 SEQID NO: 101 SEQID NO: 129

GYTFTSYG ISAYNGNT ARDSYYFDK SEQID NO: 80 SEQID NO: 102 SEQID NO: 130

GYTFTNYG ISPYNDYT ARSFYDS SEQID NO: 81 SEQID NO: 103 SEQID NO: 131

GYTFTSYG INTYNGNT ARYSSTG RWQDW SEQID NO: 80 SEQID NO: 104 SEQID NO: 132

GYSFTSYW IDPSDSYT ARIGGMVKDT SEQID NO: 76 SEQID NO: 98 SEQID NO: 133

GGTFSSYA IIPILDIP ARGYGWQSYDG SEQID NO: 82 SEQID NO: 105 SEQID NO: 134

GYTFSSYG ISTYNGNS ARSPMDY SEQID NO: 83 SEQID NO: 106 SEQID NO: 135

GYTFTDYS MNTNTGKP ARSSGDY SEQID NO: 84 SEQID NO: 107 SEQID NO: 136

GYTFTSYD MNPNSGNT ARSDFDS SEQID NO: 85 SEQID NO: 108 SEQID NO: 137

GYIFNNYY INTYTGNP ARGSYSWSYYSQKDY SEQID NO: 86 SEQID NO: 109 SEQID NO: 138

GYTFTSYG ISAYNGNT ARDAYVYDS SEQID NO: 80 SEQID NO: 102 SEQID NO: 139

GFTFSDYY ISSSGSTI ARQGWYMYLGWDY SEQID NO: 79 SEQID NO: 110 SEQID NO: 140

GLTFSMYA ISSSGGST ARGQHGSYYSYSDY SEQID NO: 87 SEQID NO: 111 SEQID NO: 141

GYTFTSYD MNPNSGNT ARMSHRVGYMGAGFDP SEQID NO: 85 SEQID NO: 108 SEQID NO: 142

GYTFTSYG MNPHSGNT ARSGFDI SEQID NO: 80 SEQID NO: 112 SEQID NO: 143

GYPFTSYG ISGYNGNT ARWWHPWYGDH SEQID NO: 88 SEQID NO: 113 SEQID NO: 144

GGTFSSYA IIPILGIA ARGSISYSIYFMSSDI SEQID NO: 82 SEQID NO: 114 SEQID NO: 145

GGSISPYY ISDSGTA ARGRSVMGYYYSDY SEQID NO: 89 SEQID NO: 115 SEQID NO: 146 23 GYTFTSYA INTNTGNP ARSHYDI SEQID NO: 90 SEQID NO: 116 SEQID NO: 147

24 GYTFTNYG IDPSGGST ARQAVDQ

SEQID NO: 81 SEQID NO: 117 SEQID NO: 148

26 GGTFSSYA IIPIFGTA ARYRGSSLWYQYVDY

SEQID NO: 82 SEQID NO: 118 SEQID NO: 149

27 GGTFSSYA IIPILGIA ARAYGRSYYDS

SEQID NO: 82 SEQID NO: 114 SEQID NO: 150

28 GYSFSTSD MNPDSGNA ARGMDY

SEQID NO: 91 SEQID NO: 119 SEQID NO: 151

30 GYNFLNYG ISTYTGNT ARSWGGYPWYSMDY

SEQID NO: 92 SEQID NO: 120 SEQID NO: 152

31 GYTFTSYG ISAYNGNT ARTWPRWTLDY

SEQID NO: 80 SEQID NO: 102 SEQID NO: 153

32 GYAFSNYW IYPGDSDT ARGFWYRDG

SEQID NO: 93 SEQID NO: 121 SEQID NO: 154

33 GYNFLNYG ISTYTGNT ARSYQDV

SEQID NO: 92 SEQID NO: 120 SEQID NO: 155

34 GYTFTSYG ISAYNGNT ARGVRSSQYDY

SEQID NO: 80 SEQID NO: 102 SEQID NO: 156

35 GYTFTSYD MNPNSGNT ARSGWDSSSGFYLDHDK

SEQID NO: 85 SEQID NO: 108 SEQID NO: 157

36 GYTFTSYD MNPNSGNT ARGGFDY

SEQID NO: 85 SEQID NO: 108 SEQID NO: 158

37 GFTFSDYY ISSSGSTI ARLFYYPHDDW

SEQID NO: 79 SEQID NO: 110 SEQID NO: 159

38 GYNFLNYG MNPNSGNT ARSDGDS

SEQID NO: 92 SEQID NO: 108 SEQID NO: 160

39 GDSVSSNSAA TYYRSKWYN ARGLWSSYGFDN

SEQID NO: 94 SEQID NO: 122 SEQID NO: 161

40 GGSFSGYY INHSGST ARYNYGSIDS

SEQID NO: 95 SEQID NO: 123 SEQID NO: 162

41 GYTFTGYY INPNSGGT ARYWYYSGMDY

SEQID NO: 96 SEQID NO: 124 SEQID NO: 163

TABLE 4

Clone # LC-CDR1 LC-CDR2 LC-CDR3

1 SSNIGAGYD YNS QSYDSSLSGYV

SEQID NO: 164 SEQID NO: 190 SEQID NO: 208

2 QTISTY AAS QQSYNTPIT

SEQID NO: 165 SEQID NO: 191 SEQID NO: 209

3 SSNIGAGYD GNS QSYDSSLSGLYV

SEQID NO: 164 SEQID NO: 192 SEQID NO: 210

4 SSDVGGYNY EVT SSYAGSKGV SEQID NO: 166 SEQID NO: 193 SEQID NO: 211

TSKLGPGYD HNS QSYDTSLSGSV SEQID NO: 167 SEQID NO: 194 SEQID NO: 212

SSDIGAGND GNS QSYDGSLSGGSQV SEQID NO: 168 SEQID NO: 192 SEQID NO: 213

QGIS W AAS QQANNFPLT SEQID NO: 169 SEQID NO: 191 SEQID NO: 214

SSNIGHNY AHN GTWDDRLSGEV SEQID NO: 170 SEQID NO: 195 SEQID NO: 215

SSNILNNA YSD AVWDDSVKGYV SEQID NO: 171 SEQID NO: 196 SEQID NO: 216

SSNIGAGYD GNS QSYDSSLSGSHYV SEQID NO: 164 SEQID NO: 192 SEQID NO: 217

QGISRW AAS QQANSFPLT SEQID NO: 169 SEQID NO: 191 SEQID NO: 218

QDINRW SAS QQAKSFPLT SEQID NO: 172 SEQID NO: 197 SEQID NO: 219

QGISRW AAS QQANSFPLT SEQID NO: 169 SEQID NO: 191 SEQID NO: 218

SSNIGSNT SNN AAWDDSLNGYV SEQID NO: 173 SEQID NO: 198 SEQID NO: 220

RSNIGAGHD SNS QSYDRSQNGPV SEQID NO: 174 SEQID NO: 199 SEQID NO: 221

SSNIGAGYD GNS QSYDSSLSGYV SEQID NO: 164 SEQID NO: 192 SEQID NO: 208

SSNIGNNY DNN GTWDSSLSAGV SEQID NO: 175 SEQID NO: 200 SEQID NO: 222

NSNIGAGYD GNS QSYDSSLSGYV SEQID NO: 176 SEQID NO: 192 SEQID NO: 208

QGISSW AAS QQANSFPIT SEQID NO: 177 SEQID NO: 191 SEQID NO: 223

SSDVGSYNL AGD SSYAGSNTFV SEQID NO: 178 SEQID NO: 201 SEQID NO: 224

QDIDDD EAT LQHDNFHTWT SEQID NO: 179 SEQID NO: 202 SEQID NO: 225

SSNIGNNY DNN GTWDSSLSAYV SEQID NO: 175 SEQID NO: 200 SEQID NO: 226

QGISRW AAS QQANTYPLT SEQID NO: 169 SEQID NO: 191 SEQID NO: 227

QGISRW AAS QQANSFPLT SEQID NO: 169 SEQID NO: 191 SEQID NO: 218

SSNIAVNG SNS AAWDDSPNAHYV SEQID NO: 180 SEQID NO: 199 SEQID NO: 228

NSNIGAGYD DNN QSYGSSLMRV SEQID NO: 176 SEQID NO: 200 SEQID NO: 229

QDISTW AAS QQANSFPFT SEQID NO: 181 SEQID NO: 191 SEQID NO: 230

NSNIGSNKD GYN AAWDDSLNGRV SEQID NO: 182 SEQID NO: 203 SEQID NO: 231 31 SSNIGAGYD GNS QSYDSSLSGSWV

SEQ ID NO: 164 SEQ ID NO: 192 SEQ ID NO: 232

32 QSVSSSY GAS QQYGSSPLT

SEQ ID NO: 183 SEQ ID NO: 204 SEQ ID NO: 233

33 QDIS W AAS QQANNFPLT

SEQ ID NO: 184 SEQ ID NO: 191 SEQ ID NO: 214

34 SSNIGAGYD GNS QSYDSSLSGHVV

SEQ ID NO: 164 SEQ ID NO: 192 SEQ ID NO: 234

35 SSNIGNYY DNN GSWDSSLSAGV

SEQ ID NO: 185 SEQ ID NO: 200 SEQ ID NO: 235

36 QDISSW AAS QQANSFPLT

SEQ ID NO: 186 SEQ ID NO: 191 SEQ ID NO: 218

37 NSNLGAGYD GNV QSYDSSLRAVI

SEQ ID NO: 187 SEQ ID NO: 205 SEQ ID NO: 236

38 QGISSW AAS QQANSFPLT

SEQ ID NO: 177 SEQ ID NO: 191 SEQ ID NO: 218

39 SSNIGSNT TNN AAWDANLNG

SEQ ID NO: 173 SEQ ID NO: 206 SEQ ID NO: 237

40 SSNIEKNY DNN GVWDGTVRVYF

SEQ ID NO: 188 SEQ ID NO: 200 SEQ ID NO: 238

41 NIGSKS DDS QVWDSSSDH

SEQ ID NO: 189 SEQ ID NO: 207 SEQ ID NO: 239

[0145] In some embodiments, the anti-RTMC antibody moiety comprises a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (including for example at least any of 96%, 97%, 98%, or 99%) sequence identity.

[0146] In some embodiments, the anti-RTMC antibody moiety comprises a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74.

[0147] The heavy and light chain variable domains can be combined in various pair-wise combinations to generate a number of anti-RTMC antibody moieties.

[0148] For example, in some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC- CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively, SEQ ID NOs: 77, 99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and 214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105, 134, 164, 192, and 217, respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218, respectively, SEQ ID NOs: 84, 107, 136, 172, 197, and 219, respectively, SEQ ID NOs: 85, 108, 137, 169, 191, and 218,

respectively, SEQ ID NOs: 86, 109, 138, 173, 198, and 220, respectively, SEQ ID NOs: 80, 102, 139, 174, 199, and 221, respectively, SEQ ID NOs: 79, 110, 140, 164, 192, and 208,

respectively, SEQ ID NOs: 87, 111, 141, 175, 200, and 222, respectively, SEQ ID NOs: 85, 108, 142, 176, 192, and 208, respectively, SEQ ID NOs: 80, 112, 143, 177, 191, and 223,

respectively, SEQ ID NOs: 88, 113, 144, 178, 201, and 224, respectively, SEQ ID NOs: 82, 114, 145, 179, 202, and 225, respectively, SEQ ID NOs: 89, 115, 146, 175, 200, and 226,

respectively, SEQ ID NOs: 90, 116, 147, 169, 191, and 227, respectively, SEQ ID NOs: 81, 117, 148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118, 149, 180, 199, and 228,

respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229, respectively, SEQ ID NOs: 91, 119, 151, 181, 191, and 230, respectively, SEQ ID NOs: 92, 120, 152, 182, 203, and 231,

respectively, SEQ ID NOs: 80, 102, 153, 164, 192, and 232, respectively, SEQ ID NOs: 93, 121, 154, 183, 204, and 233, respectively, SEQ ID NOs: 92, 120, 155, 184, 191, and 214,

respectively, SEQ ID NOs: 80, 102, 156, 164, 192, and 234, respectively, SEQ ID NOs: 85, 108, 157, 185, 200, and 235, respectively, SEQ ID NOs: 85, 108, 158, 186, 191, and 218,

respectively, SEQ ID NOs: 79, 110, 159, 187, 205, and 236, respectively, SEQ ID NOs: 92, 108, 160, 177, 191, and 218, respectively, SEQ ID NOs: 94, 122, 161, 173, 206, and 237,

respectively, SEQ ID NOs: 95, 123, 162, 188, 200, and 238, respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively; or variants thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions, individually, in HC-CDR1, HC- CDR2, HC-CDR3, LC-CDR1, and/or LC-CDR3, and/or up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions in LC-CDR2. For example, in some embodiments, the heavy chain comprises an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 75, an HC- CDR2 comprising the amino acid sequence of SEQ ID NO: 97, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO 125, and the light chain comprises an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 164, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 190, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 208. [0149] In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and

208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively, SEQ ID NOs: 77,

99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102,

130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and 214,

respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76, 98,

133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105, 134, 164, 192, and 217,

respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218, respectively, SEQ ID NOs: 84, 107,

136, 172, 197, and 219, respectively, SEQ ID NOs: 85, 108, 137, 169, 191, and 218,

respectively, SEQ ID NOs: 86, 109, 138, 173, 198, and 220, respectively, SEQ ID NOs: 80, 102,

139, 174, 199, and 221, respectively, SEQ ID NOs: 79, 110, 140, 164, 192, and 208,

respectively, SEQ ID NOs: 87, 111, 141, 175, 200, and 222, respectively, SEQ ID NOs: 85, 108,

142, 176, 192, and 208, respectively, SEQ ID NOs: 80, 112, 143, 177, 191, and 223,

respectively, SEQ ID NOs: 88, 113, 144, 178, 201, and 224, respectively, SEQ ID NOs: 82, 114,

145, 179, 202, and 225, respectively, SEQ ID NOs: 89, 115, 146, 175, 200, and 226,

respectively, SEQ ID NOs: 90, 116, 147, 169, 191, and 227, respectively, SEQ ID NOs: 81, 117,

148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118, 149, 180, 199, and 228,

respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229, respectively, SEQ ID NOs: 91, 119,

151, 181, 191, and 230, respectively, SEQ ID NOs: 92, 120, 152, 182, 203, and 231,

respectively, SEQ ID NOs: 80, 102, 153, 164, 192, and 232, respectively, SEQ ID NOs: 93, 121,

154, 183, 204, and 233, respectively, SEQ ID NOs: 92, 120, 155, 184, 191, and 214,

respectively, SEQ ID NOs: 80, 102, 156, 164, 192, and 234, respectively, SEQ ID NOs: 85, 108,

157, 185, 200, and 235, respectively, SEQ ID NOs: 85, 108, 158, 186, 191, and 218,

respectively, SEQ ID NOs: 79, 110, 159, 187, 205, and 236, respectively, SEQ ID NOs: 92, 108,

160, 177, 191, and 218, respectively, SEQ ID NOs: 94, 122, 161, 173, 206, and 237,

respectively, SEQ ID NOs: 95, 123, 162, 188, 200, and 238, respectively, or SEQ ID NOs: 96,

124, 163, 189, 207, and 239, respectively, or variants thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences and/or up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences.

[0150] In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively, SEQ ID NOs: 77, 99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and 214,

respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105, 134, 164, 192, and 217,

respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218, respectively, SEQ ID NOs: 84, 107, 136, 172, 197, and 219, respectively, SEQ ID NOs: 85, 108, 137, 169, 191, and 218,

respectively, SEQ ID NOs: 95, 123, 162, 188, 200, and 238, respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively.

[0151] In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ

ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and

52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively,

SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively. The sequences of the heavy chain variable domains and light chain variable domains from putative anti-RTMC antibody clones are provided in Table 5 below.

TABLE 5

Clone # Heavy chain variable domain Light chain variable domain

(SEQ ID NO) (SEQ ID NO)

1 19 47

3 20 48

4 21 49

5 22 50

6 23 51

7 24 52

8 25 53

9 26 54

10 27 55 12 28 56

13 29 57

14 30 58

15 31 59

16 32 60

17 33 61

18 34 62

19 35 63

20 36 64

21 37 65

22 38 66

24 39 67

26 40 68

27 41 69

30 42 70

32 43 71

34 44 72

37 45 73

39 46 74

[0152] In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

[0153] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target HIV-1 RT/MHC class I complex with a second anti-RTMC antibody moiety according to any of the anti-RTMC antibody moieties described herein. In some embodiments, the anti-RTMC antibody moiety binds to the same, or substantially the same, epitope as the second anti-RTMC antibody moiety. In some embodiments, binding of the anti-RTMC antibody moiety to the target HIV-1 RT/MHC class I complex inhibits binding of the second anti-RTMC antibody moiety to the target HIV-1 RT/MHC class I complex by at least about 70% (such as by at least about any of 75%, 80%, 85%, 90%, 95%, 98% or 99%), or vice versa. In some embodiments, the anti- RTMC antibody moiety and the second anti-RTMC antibody moiety cross-compete for binding to the target HIV-1 RT/MHC class I complex, i.e., each of the antibody moieties competes with the other for binding to the target HIV-1 RT/MHC class I complex.

[0154] For example, in some embodiments, the anti-RTMC antibody moiety competes for binding to a target HIV-1 RT/MHC class I complex with an antibody moiety comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247- 249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions.

[0155] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target

HIV-1 RT/MHC class I complex with an antibody moiety comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an HC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of

SEQ ID NOs: 97-124; or a variant thereof comprising up to about 5 (for example about any of 1,

2, 3, 4, or 5) amino acid substitutions; and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207; or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and an LC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions.

[0156] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target HIV-1 RT/MHC class I complex with an antibody moiety comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 97- 124; and an HC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences.

[0157] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target HIV-1 RT/MHC class I complex with an antibody moiety comprising a heavy chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising (and in some embodiments consisting of) the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity.

[0158] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target

HIV-1 RT/MHC class I complex with an antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and

99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and 214,

respectively, SEQ ID NOs: 95, 123, 162, 188, 200, and 238, respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively; or variants thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions, individually, in HC-CDR1, HC- CDR2, HC-CDR3, LC-CDR1, and/or LC-CDR3, and/or up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions in LC-CDR2.

[0159] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target

[0160] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target

[0161] In some embodiments, the anti-RTMC antibody moiety competes for binding to a target

HIV-1 RT/MHC class I complex with an antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ

SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ

ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC antibody moiety competes for binding to a target HIV-1 RT/MHC class I complex with an antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively.

Full-length anti-RTMC antibodies

[0162] The anti-RTMC constructs in some embodiments are full-length antibodies comprising an anti-RTMC antibody moiety (also referred to herein as a "full-length anti-RTMC antibody"). In some embodiments, the full-length antibody is a monoclonal antibody.

[0163] In some embodiments, the full-length anti-RTMC antibody comprises an Fc sequence from an immunoglobulin, such as IgA, IgD, IgE, IgG, and IgM. In some embodiments, the full- length anti-RTMC antibody comprises an Fc sequence of IgG, such as any of IgGl, IgG2, IgG3, or IgG4. In some embodiments, the full-length anti-RTMC antibody comprises an Fc sequence of a human immunoglobulin. In some embodiments, the full-length anti-RTMC antibody comprises an Fc sequence of a mouse immunoglobulin. In some embodiments, the full-length anti-RTMC antibody comprises an Fc sequence that has been altered or otherwise changed so that it has enhanced antibody dependent cellular cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC) effector function.

[0164] Thus, for example, in some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, and b) an Fc region. In some embodiments, the HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, and b) an Fc region. In some embodiments, the Fc region comprises an IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence. In some embodiments, the Fc region comprises a mouse IgGl Fc sequence. In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0165] In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID

NOs: 245-246; or a variant thereof comprising up to about 3 (for example about any of 1, 2, or

3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and b) an Fc region. In some embodiments, the Fc region comprises an IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence. In some embodiments, the Fc region comprises a mouse IgGl Fc sequence.

[0166] In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an HC-

CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-

CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of

SEQ ID NOs: 250-253, and b) an Fc region. In some embodiments, the Fc region comprises an

IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence.

In some embodiments, the Fc region comprises a mouse IgGl Fc sequence.

[0167] In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an

HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID

NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or

5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-

CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the Fc region comprises an IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence. In some embodiments, the Fc region comprises a mouse IgGl Fc sequence.

[0168] In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75- 96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences; and b) an Fc region. In some embodiments, the Fc region comprises an IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence. In some embodiments, the Fc region comprises a mouse IgGl Fc sequence.

[0169] In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75- 96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC-CDR 1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; and b) an Fc region. In some embodiments, the Fc region comprises an IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence. In some embodiments, the Fc region comprises a mouse IgGl Fc sequence.

[0170] In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47- 74, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; and b) an Fc region. In some embodiments, the Fc region comprises an IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence. In some embodiments, the Fc region comprises a mouse IgGl Fc sequence.

[0171] In some embodiments, there is provided a full-length anti-RTMC antibody comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74; and b) an Fc region. In some embodiments, the Fc region comprises an IgGl Fc sequence. In some embodiments, the Fc region comprises a human IgGl Fc sequence. In some embodiments, the Fc region comprises a mouse IgGl Fc sequence.

[0172] For example, in some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-

CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID

NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively, SEQ ID NOs: 77, 99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100,

128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212,

respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103,

131, 169, 191, and 214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215,

respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105,

134, 164, 192, and 217, respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218,

respectively, SEQ ID NOs: 84, 107, 136, 172, 197, and 219, respectively, SEQ ID NOs: 85, 108,

137, 169, 191, and 218, respectively, SEQ ID NOs: 86, 109, 138, 173, 198, and 220,

respectively, SEQ ID NOs: 80, 102, 139, 174, 199, and 221, respectively, SEQ ID NOs: 79, 110,

140, 164, 192, and 208, respectively, SEQ ID NOs: 87, 111, 141, 175, 200, and 222,

respectively, SEQ ID NOs: 85, 108, 142, 176, 192, and 208, respectively, SEQ ID NOs: 80, 112,

143, 177, 191, and 223, respectively, SEQ ID NOs: 88, 113, 144, 178, 201, and 224,

respectively, SEQ ID NOs: 82, 114, 145, 179, 202, and 225, respectively, SEQ ID NOs: 89, 115, 146, 175, 200, and 226, respectively, SEQ ID NOs: 90, 116, 147, 169, 191, and 227,

respectively, SEQ ID NOs: 81, 117, 148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118,

149, 180, 199, and 228, respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229,

respectively, SEQ ID NOs: 91, 119, 151, 181, 191, and 230, respectively, SEQ ID NOs: 92, 120,

152, 182, 203, and 231, respectively, SEQ ID NOs: 80, 102, 153, 164, 192, and 232,

respectively, SEQ ID NOs: 93, 121, 154, 183, 204, and 233, respectively, SEQ ID NOs: 92, 120,

155, 184, 191, and 214, respectively, SEQ ID NOs: 80, 102, 156, 164, 192, and 234,

respectively, SEQ ID NOs: 85, 108, 157, 185, 200, and 235, respectively, SEQ ID NOs: 85, 108,

158, 186, 191, and 218, respectively, SEQ ID NOs: 79, 110, 159, 187, 205, and 236,

respectively, SEQ ID NOs: 92, 108, 160, 177, 191, and 218, respectively, SEQ ID NOs: 94, 122,

161, 173, 206, and 237, respectively, SEQ ID NOs: 95, 123, 162, 188, 200, and 238,

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively; or variants thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions, individually, in HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, and/or LC-CDR3, and/or up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions in LC-CDR2.

[0173] In some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC-

CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97,

125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively,

SEQ ID NOs: 77, 99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100, 128, 166,

193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID

NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and

214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76,

98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105, 134, 164, 192, and 217, respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218, respectively, SEQ ID NOs: 84, 107,

respectively, SEQ ID NOs: 90, 116, 147, 169, 191, and 227, respectively, SEQ ID NOs: 81, 117, 148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118, 149, 180, 199, and 228, respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229, respectively, SEQ ID NOs: 91, 119, 151, 181, 191, and 230, respectively, SEQ ID NOs: 92, 120, 152, 182, 203, and 231,

[0174] In some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC- CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97,

[0175] In some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ

ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and

54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively,

SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ

ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and

66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively,

SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity.

In some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ

ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively.

[0176] In some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the full- length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the full-length anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

[0177] In some embodiments, the full-length anti-RTMC antibody binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein with a IQ between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the full-length anti-RTMC antibody binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein with a K_d between about 1 pM to about 250 pM (such as about any of 1, 10, 25, 50, 75, 100, 150, 200, or 250 pM, including any ranges between these values).

Multi-Specific anti-RTMC molecules

[0178] The anti-RTMC constructs in some embodiments comprise a multi- specific anti-RTMC molecule comprising an anti-RTMC antibody moiety and a second binding moiety (such as a second antigen-binding moiety). In some embodiments, the multi- specific anti-RTMC molecule comprises an anti-RTMC antibody moiety and a second antigen-binding moiety.

[0179] Multi- specific molecules are molecules that have binding specificities for at least two different antigens or epitopes (e.g., bispecific antibodies have binding specificities for two antigens or epitopes). Multi- specific molecules with more than two valencies and/or specificities are also contemplated. For example, trispecific antibodies can be prepared. Tutt et al. J.

Immunol. 147: 60 (1991). It is to be appreciated that one of skill in the art could select appropriate features of individual multi- specific molecules described herein to combine with one another to form a multi- specific anti-RTMC molecule of the invention.

[0180] Thus, for example, in some embodiments, there is provided a multi- specific {e.g., bispecific) anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, and b) a second binding moiety (such as an antigen-binding moiety). In some embodiments, the second binding moiety specifically binds to a complex comprising a different HIV-1 RT peptide bound to the MHC class I protein. In some embodiments, the second scFv specifically binds to a complex comprising the HIV-1 RT peptide bound to a different MHC class I protein. In some embodiments, the second binding moiety specifically binds to a different epitope on the complex comprising the HIV-1 RT peptide and the MHC class I protein. In some embodiments, the second binding moiety specifically binds to a different antigen. In some embodiments, the second binding moiety specifically binds to an antigen on the surface of a cell, such as a cytotoxic cell. In some embodiments, the second binding moiety specifically binds to an antigen on the surface of a lymphocyte, such as a T cell, an NK cell, a neutrophil, a monocyte, a macrophage, or a dendritic cell. In some embodiments, the second binding moiety specifically binds to an effector T cell, such as a cytotoxic T cell (also known as cytotoxic T lymphocyte (CTL) or T killer cell).

[0181] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, and b) a second antigen-binding moiety that binds specifically to CD3. In some embodiments, the second antigen-binding moiety specifically binds to CD3s. In some embodiments, the second antigen-binding moiety specifically binds to an agonistic epitope of CD3s. The term "agonistic epitope", as used herein, means (a) an epitope that, upon binding of the multi- specific molecule, optionally upon binding of several multi- specific molecules on the same cell, allows said multi- specific molecules to activate T cell receptor (TCR) signaling and induce T cell activation, and/or (b) an epitope that is solely composed of amino acid residues of the epsilon chain of CD3 and is accessible for binding by the multi- specific molecule, when presented in its natural context on T cells {i.e. surrounded by the TCR, the CD3y chain, etc.), and/or (c) an epitope that, upon binding of the multi- specific molecule, does not lead to stabilization of the spatial position of CD3s relative to CD3y.

[0182] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an

HIV- 1 RT peptide and an MHC class I protein, and b) a second antigen-binding moiety that binds specifically to an antigen on the surface of an effector cell, including for example CD3y,

CD35, CD38, CD3C, CD28, CD16a, CD56, CD68, and GDS2D.

[0183] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an

HIV- 1 RT peptide and an MHC class I protein, and b) a second antigen-binding moiety that binds specifically to a component of the complement system, such as Clq. Clq is a subunit of the CI enzyme complex that activates the serum complement system.

[0184] In some embodiments, the second antigen-binding moiety specifically binds to an Fc receptor. In some embodiments, the second antigen-binding moiety specifically binds to an Fey receptor (FcyR). The FcyR may be an FcyRIII present on the surface of natural killer (NK) cells or one of FcyRI, FcyRIIA, FcyRIIBI, FcyRIIB2, and FcyRIIIB present on the surface of macrophages, monocytes, neutrophils and/or dendritic cells. In some embodiments, the second antigen-binding moiety is an Fc region or functional fragment thereof. A "functional fragment" as used in this context refers to a fragment of an antibody Fc region that is still capable of binding to an FcR, in particular to an FcyR, with sufficient specificity and affinity to allow an

FcyR bearing effector cell, in particular a macrophage, a monocyte, a neutrophil and/or a dendritic cell, to kill the target cell by cytotoxic lysis or phagocytosis. A functional Fc fragment is capable of competitively inhibiting the binding of the original, full-length Fc portion to an

FcR such as the activating FcyRI. In some embodiments, a functional Fc fragment retains at least

30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% of its affinity to an activating FcyR. In some embodiments, the Fc region or functional fragment thereof is an enhanced Fc region or functional fragment thereof. The term "enhanced Fc region", as used herein, refers to an Fc region that is modified to enhance Fc receptor-mediated effector-functions, in particular antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity

(CDC), and antibody-mediated phagocytosis. This can be achieved as known in the art, for example by altering the Fc region in a way that leads to an increased affinity for an activating receptor (e.g. FcyRIIIA (CD 16 A) expressed on natural killer (NK) cells) and/or a decreased binding to an inhibitory receptor (e.g. FcyRIIB l/B2 (CD32B)). In yet other embodiments, the second antigen-binding moiety is an antibody or antigen-binding fragment thereof that specifically binds to an FcR, in particular to an FcyR, with sufficient specificity and affinity to allow an FcyR bearing effector cell, in particular a macrophage, a monocyte, a neutrophil and/or a dendritic cell, to kill the target cell by cytotoxic lysis or phagocytosis.

[0185] In some embodiments, the multi- specific anti-RTMC molecule allows killing of RTMC- presenting target cells and/or can effectively redirect CTLs to lyse RTMC-presenting target cells. In some embodiments, the multi- specific (e.g. , bispecific) anti-RTMC molecule of the present invention shows an in vitro EC₅₀ ranging from 10 to 500 ng/ml, and is able to induce redirected lysis of about 50% of the target cells through CTLs at a ratio of CTLs to target cells of from about 1 : 1 to about 50: 1 (such as from about 1: 1 to about 15: 1, or from about 2: 1 to about 10: 1).

[0186] In some embodiments, the multi- specific (e.g. , bispecific) anti-RTMC molecule is capable of cross-linking a stimulated or unstimulated CTL and the target cell in such a way that the target cell is lysed. This offers the advantage that no generation of target- specific T cell clones or common antigen presentation by dendritic cells is required for the multi- specific anti- RTMC molecule to exert its desired activity. In some embodiments, the multi- specific anti- RTMC molecule of the present invention is capable of redirecting CTLs to lyse the target cells in the absence of other activating signals. In some embodiments, the second antigen-binding moiety of the multi- specific anti-RTMC molecule specifically binds to CD3 (e.g., specifically binds to CD3s), and signaling through CD28 and/or IL-2 is not required for redirecting CTLs to lyse the target cells.

[0187] Methods for measuring the preference of the multi- specific anti-RTMC molecule to simultaneously bind to two antigens (e.g. , antigens on two different cells) are within the normal capabilities of a person skilled in the art. For example, when the second binding moiety specifically binds to CD3, the multi- specific anti-RTMC molecule may be contacted with a mixture of CD3⁺/HIV- 1 RT^" cells and CD37HIV- 1 RT⁺ cells. The number of multi- specific anti- RTMC molecule-positive single cells and the number of cells cross-linked by multi- specific anti-RTMC molecules may then be assessed by microscopy or fluorescence-activated cell sorting (FACS) as known in the art.

[0188] For example, in some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, and b) a second antigen-binding moiety. In some embodiments, the HIV- 1 RT peptide is HIV- 1 RT 181 (SEQ ID NO: 5), HIV- 1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the second antigen-binding moiety specifically binds to a complex comprising a different HIV-1 RT peptide bound to the MHC class I protein. In some embodiments, the second antigen-binding moiety specifically binds to a complex comprising the HIV-1 RT peptide bound to a different MHC class I protein. In some

embodiments, the second antigen-binding moiety specifically binds to a different epitope on the complex comprising the HIV-1 RT peptide and the MHC class I protein. In some embodiments, the second antigen-binding moiety specifically binds to another antigen. In some embodiments, the second antigen-binding moiety specifically binds to an antigen on the surface of a cell, such as an RTMC-presenting cell. In some embodiments, the second antigen-binding moiety specifically binds to an antigen on the surface of a cell that does not express HIV-1 RT. In some embodiments, the second antigen-binding moiety specifically binds to an antigen on the surface of a cytotoxic cell. In some embodiments, the second antigen-binding moiety specifically binds to an antigen on the surface of a lymphocyte, such as a T cell, an NK cell, a neutrophil, a monocyte, a macrophage, or a dendritic cell. In some embodiments, the second antigen-binding moiety specifically binds to an antigen on the surface of an effector T cell, such as a cytotoxic T cell. In some embodiments, the second antigen-binding moiety specifically binds to an antigen on the surface of an effector cell, including for example CD3y, CD35, CD3s, CD3ζ, CD28, CD16a, CD56, CD68, and GDS2D. In some embodiments, the anti-RTMC antibody moiety is human, humanized, or semi- synthetic. In some embodiments, the second antigen-binding moiety is an antibody moiety. In some embodiments, the second antigen-binding moiety is a human, humanized, or semi- synthetic antibody moiety. In some embodiments, the multi- specific anti- RTMC molecule further comprises at least one (such as at least about any of 2, 3, 4, 5, or more) additional antigen-binding moieties. In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0189] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, and b) a second antigen-binding moiety.

[0190] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and b) a second antigen-binding moiety.

[0191] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an

HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an

HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an

HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, and b) a second antigen-binding moiety.

[0192] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an

HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-

96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid

substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and b) a second antigen-binding moiety.

[0193] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97- 124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125- 163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences; and b) a second antigen-binding moiety.

[0194] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97- 124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125- 163; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; and b) a second antigen-binding moiety.

[0195] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; and b) a second scFv.

[0196] In some embodiments, there is provided a multi- specific anti-RTMC molecule comprising a) an anti-RTMC antibody moiety comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74; and b) a second antigen-binding moiety.

[0197] In some embodiments, the multi- specific anti-RTMC molecule is, for example, a diabody (Db), a single-chain diabody (scDb), a tandem scDb (Tandab), a linear dimeric scDb (LD-scDb), a circular dimeric scDb (CD-scDb), a di-diabody, a tandem scFv, a tandem di-scFv (e.g. , a bispecific T cell engager), a tandem tri-scFv, a tri(a)body, a bispecific Fab2, a di-miniantibody, a tetrabody, an scFv-Fc-scFv fusion, a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, an IgG-scFab, an scFab-ds-scFv, an Fv2-Fc, an IgG-scFv fusion, a dock and lock (DNL) antibody, a knob-into-hole (KiH) antibody (bispecific IgG prepared by the KiH technology), a DuoBody (bispecific IgG prepared by the Duobody technology), a heteromultimeric antibody, or a heteroconjugate antibody. In some embodiments, the multi- specific anti-RTMC molecule is a tandem scFv (e.g. , a tandem di-scFv, such as a bispecific T cell engager).

Tandem scFv

[0198] The multi- specific anti-RTMC molecule in some embodiments is a tandem scFv comprising a first scFv comprising an anti-RTMC antibody moiety and a second scFv (also referred to herein as a "tandem scFv multi- specific anti-RTMC antibody"). In some

embodiments, the tandem scFv multi- specific anti-RTMC antibody further comprises at least one (such as at least about any of 2, 3, 4, 5, or more) additional scFv. [0199] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, and b) a second scFv. In some embodiments, the HIV- 1 RT peptide is HIV- 1 RT 181 (SEQ ID NO: 5), HIV- 1 RT 181 M184V (SEQ ID NO: 6), HIV- 1 RT 181 M184I (SEQ ID NO: 7), HIV- 1 RT 181 Y181C (SEQ ID NO: 8), or HIV- 1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the second scFv specifically binds to a complex comprising a different HIV- 1 RT peptide bound to the MHC class I protein. In some embodiments, the second scFv specifically binds to a complex comprising the HIV- 1 RT peptide bound to a different MHC class I protein. In some embodiments, the second scFv specifically binds to a different epitope on the complex comprising the HIV- 1 RT peptide and the MHC class I protein. In some embodiments, the second scFv specifically binds to another antigen. In some embodiments, the second scFv specifically binds to an antigen on the surface of a cell, such as an RTMC-presenting cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of a cell that does not express HIV- 1 RT. In some embodiments, the second scFv specifically binds to an antigen on the surface of a cytotoxic cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of a lymphocyte, such as a T cell, an NK cell, a neutrophil, a monocyte, a macrophage, or a dendritic cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of an effector T cell, such as a cytotoxic T cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of an effector cell, including for example CD3y, CD35, CD3s, ΟΌ3ζ, CD28, CD16a, CD56, CD68, and GDS2D. In some embodiments, the first scFv is human, humanized, or semi- synthetic. In some

embodiments, the second scFv is human, humanized, or semi- synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi- synthetic. In some embodiments, the tandem scFv multi- specific anti-RTMC antibody further comprises at least one (such as at least about any of 2, 3, 4, 5, or more) additional scFv. In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV- 1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV- 1 RT peptide and a different subtype of the MHC class I protein. [0200] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT 181 (SEQ ID NO: 5), FflV- 1 RT 181 M184V (SEQ ID NO: 6), FflV- 1 RT 181 M184I (SEQ ID NO: 7), HIV- 1 RT 181 Y181C (SEQ ID NO: 8), or HIV- 1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, and b) a second scFv.

[0201] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and b) a second scFv.

[0202] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, and b) a second scFv.

[0203] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75- 96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97- 124, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125- 163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid

substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and b) a second scFv.

[0204] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97- 124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125- 163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences; and b) a second scFv.

[0205] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID

NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-

124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-

163; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; and b) a second scFv.

[0206] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; and b) a second scFv.

[0207] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74; and b) a second scFv.

[0208] In some embodiments, there is provided a tandem scFv multi- specific (e.g. , bispecific) anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, and b) a second scFv, wherein the tandem scFv multi- specific anti-RTMC antibody is a tandem di-scFv or a tandem tri-scFv. In some embodiments, the tandem scFv multi- specific anti-RTMC antibody is a tandem di-scFv. In some embodiments, the tandem scFv multi- specific anti-RTMC antibody is a bispecific T-cell engager.

[0209] For example, in some embodiments, there is provided a tandem di-scFv bispecific anti- RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, and b) a second scFv that specifically binds to an antigen on the surface of a T cell. In some embodiments, the HIV- 1 RT peptide is HIV- 1 RT 181 (SEQ ID NO: 5), HIV- 1 RT 181 M184V (SEQ ID NO: 6), HIV- 1 RT 181 M184I (SEQ ID NO: 7), HIV- 1 RT 181 Y181C (SEQ ID NO: 8), or HIV- 1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the second scFv specifically binds to an antigen on the surface of an effector T cell, such as a cytotoxic T cell. In some embodiments, the second scFv specifically binds to an antigen selected, for example, from the group consisting of

CD3y, CD35, CD3s, ΟΌ3ζ, CD28, OX40, GITR, CD137, CD27, CD40L, and HVEM. In some embodiments, the second scFv specifically binds to an agonistic epitope on an antigen on the surface of a T cell, wherein the binding of the second scFv to the antigen enhances T cell activation. In some embodiments, the first scFv is human, humanized, or semi- synthetic. In some embodiments, the second scFv is human, humanized, or semi- synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi- synthetic.

[0210] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0211] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID

NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or

3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an

LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0212] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an HC-

CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0213] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC- CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0214] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75- 96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences, and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0215] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75- 96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC-CDR 1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0216] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47- 74, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0217] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, and b) a second scFv that specifically binds to an antigen on the surface of a T cell.

[0218] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, and b) a second scFv that specifically binds to CD3s. In some embodiments, the HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181

M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ

ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the first scFv is fused to the second scFv through linkage with a peptide linker. In some embodiments, the peptide linker is between about 5 to about 20 (such as about any of 5, 10, 15, or 20, including any ranges between these values) amino acids in length. In some embodiments, the peptide linker comprises (and in some embodiments consists of) the amino acid sequence S RGGGGS GGGGSGGGGSLEMA (SEQ ID NO: 276). In some embodiments, the first scFv is human, humanized, or semi- synthetic. In some embodiments, the second scFv is human, humanized, or semi- synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi- synthetic.

[0219] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, and b) a second scFv that specifically binds to CD3s. In some embodiments, the first scFv is fused to the second scFv through linkage with a peptide linker. In some embodiments, the peptide linker is between about 5 to about 20 (such as about any of 5, 10, 15, or 20, including any ranges between these values) amino acids in length. In some embodiments, the peptide linker comprises (and in some embodiments consists of) the amino acid sequence of SEQ ID NO: 276. In some embodiments, the first scFv is human, humanized, or semi- synthetic. In some embodiments, the second scFv is human, humanized, or semi- synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi- synthetic.

[0220] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID

3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and b) a second scFv that specifically binds to CD3s. In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, and b) a second scFv that specifically binds to CD3s. In some embodiments, the first scFv is fused to the second scFv through linkage with a peptide linker. In some embodiments, the peptide linker is between about 5 to about 20 (such as about any of 5, 10, 15, or 20, including any ranges between these values) amino acids in length. In some embodiments, the peptide linker comprises (and in some embodiments consists of) the amino acid sequence of SEQ ID NO: 276. In some embodiments, the first scFv is human, humanized, or semi- synthetic. In some embodiments, the second scFv is human, humanized, or semi- synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi- synthetic.

[0221] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an

5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-

CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and b) a second scFv that specifically binds to CD3s. In some embodiments, here is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences, and b) a second scFv that specifically binds to CD3s. In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; and b) a second scFv that specifically binds to CD3s. In some embodiments, the first scFv is fused to the second scFv through linkage with a peptide linker. In some embodiments, the peptide linker is between about 5 to about 20 (such as about any of 5, 10, 15, or 20, including any ranges between these values) amino acids in length. In some embodiments, the peptide linker comprises (and in some embodiments consists of) the amino acid sequence of SEQ ID NO: 276. In some embodiments, the first scFv is human, humanized, or semi- synthetic. In some embodiments, the second scFv is human, humanized, or semi- synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi- synthetic. [0222] In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47- 74, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and b) a second scFv that specifically binds to CD3s. In some embodiments, there is provided a tandem di-scFv bispecific anti-RTMC antibody comprising a) a first scFv comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, and b) a second scFv that specifically binds to CD3s. In some embodiments, the first scFv is fused to the second scFv through linkage with a peptide linker. In some embodiments, the peptide linker is between about 5 to about 20 (such as about any of 5, 10, 15, or 20, including any ranges between these values) amino acids in length. In some embodiments, the peptide linker comprises (and in some embodiments consists of) the amino acid sequence of SEQ ID NO: 276. In some embodiments, the first scFv is human, humanized, or semi- synthetic. In some embodiments, the second scFv is human, humanized, or semi- synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi- synthetic.

[0223] In some embodiments, the tandem di-scFv bispecific anti-RTMC antibody binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein with a K_d between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the tandem di-scFv bispecific anti-RTMC antibody binds to a complex

comprising an HIV-1 RT peptide and an MHC class I protein with a IQ between about 1 nM to about 500 nM (such as about any of 1, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 nM, including any ranges between these values).

[0224] In some embodiments, the tandem di-scFv bispecific anti-RTMC antibody is stable in a solution for at least about 1 month (such as at least about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, or longer). Stability can be expressed, for example, as the retention of target cell-killing activity by the tandem di-scFv bispecific anti-RTMC antibody in an aqueous formulation kept at a storage temperature, e.g., 4° C [0225] For example, in some embodiments, the tandem di-scFv bispecific anti-RTMC antibody retains at least 40% (such as at least about any of 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or greater) target cell-killing activity in an aqueous formulation kept at a storage temperature for at least about 1 month (such as at least about any of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, or longer). In some embodiments, the storage temperature is no greater than about 25° C (such as no greater than about any of 20, 18, 16, 14, 12, 10, 8, 6, 5, 4, 3, 2, 1, or 0° C, or less). In some embodiments, the tandem di-scFv bispecific anti-RTMC antibody retains at least 60% target cell-killing activity in an aqueous formulation kept at a storage temperature of 4° C for at least about 2 years. In some embodiments, the tandem di-scFv bispecific anti-RTMC antibody comprises a) a first scFv that specifically binds to a complex comprising an HIV-1 RT 181 peptide and an MHC class I protein, and b) a second scFv that specifically binds to CD3s.

[0226] For example, in some embodiments, the multi- specific anti-RTMC molecule (such as di- scFv) comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2, and LC-

CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively, SEQ ID NOs: 77, 99,

127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100, 128, 166, 193, and 211,

respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102,

148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118, 149, 180, 199, and 228, respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229, respectively, SEQ ID NOs: 91, 119, 151, 181, 191, and 230, respectively, SEQ ID NOs: 92, 120, 152, 182, 203, and 231,

124, 163, 189, 207, and 239, respectively; or variants thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions, individually, in HC-CDR1, HC- CDR2, HC-CDR3, LC-CDR1, and/or LC-CDR3, and/or up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions in LC-CDR2.

[0227] In some embodiments, the multi- specific anti-RTMC molecule (such as di-scFv) comprises an anti-RTMC antibody moiety comprising HC-CDR1, HC-CDR2, HC-CDR3, LC- CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97,

respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229, respectively, SEQ ID NOs: 91, 119, 151, 181, 191, and 230, respectively, SEQ ID NOs: 92, 120, 152, 182, 203, and 231, respectively, SEQ ID NOs: 80, 102, 153, 164, 192, and 232, respectively, SEQ ID NOs: 93, 121, 154, 183, 204, and 233, respectively, SEQ ID NOs: 92, 120, 155, 184, 191, and 214,

[0228] In some embodiments, the multi- specific anti-RTMC molecule (such as di-scFv) comprises an anti-RTMC antibody moiety comprising HC-CDR1, HC-CDR2, HC-CDR3, LC- CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97,

[0229] In some embodiments, the multi- specific anti-RTMC molecule (such as di-scFv) comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the multi- specific anti-RTMC molecule comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ

ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively.

[0230] In some embodiments, the multi- specific anti-RTMC molecule (such as di-scFv) comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the multi- specific anti-RTMC molecule (such as di-scFv) comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the multi- specific anti-RTMC molecule (such as di- scFv) comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the multi- specific anti- RTMC molecule (such as di-scFv) comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

Chimeric Receptors and effector cells

[0231] The anti-RTMC construct in some embodiments is a chimeric receptor comprising an anti-RTMC antibody moiety (also referred to herein as an "anti-RTMC chimeric receptor"). Also provided is an effector cell (e.g., T cell) comprising a chimeric receptor comprising an anti- RTMC antibody moiety (also referred to herein as an "anti-RTMC chimeric receptor effector cell", e.g., "anti-RTMC chimeric receptor T cell").

Chimeric Antigen Receptor

[0232] In some embodiments, the chimeric receptor is a chimeric antigen receptor (CAR), and the anti-RTMC chimeric receptor is an anti-RTMC CAR. In some embodiments, the anti-RTMC CAR comprises a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein and b) an intracellular signaling domain. A transmembrane domain may be present between the extracellular domain and the intracellular domain. [0233] Between the extracellular domain and the transmembrane domain of the anti-RTMC CAR, or between the intracellular domain and the transmembrane domain of the anti-RTMC CAR, there may be a spacer domain. The spacer domain can be any oligo- or polypeptide that functions to link the transmembrane domain to the extracellular domain or the intracellular domain in the polypeptide chain. A spacer domain may comprise up to about 300 amino acids, including for example about 10 to about 100, or about 25 to about 50 amino acids.

[0234] The transmembrane domain may be derived either from a natural or from a synthetic source. Where the source is natural, the domain may be derived from any membrane-bound or transmembrane protein. Transmembrane regions of particular use in this invention may be derived from (i.e. comprise at least the transmembrane region(s) of) the α, β, δ, γ, or ζ chain of the T-cell receptor, CD28, CD3s, CD3 CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. In some embodiments, the

transmembrane domain may be synthetic, in which case it may comprise predominantly hydrophobic residues such as leucine and valine. In some embodiments, a triplet of

phenylalanine, tryptophan and valine may be found at each end of a synthetic transmembrane domain. In some embodiments, a short oligo- or polypeptide linker, having a length of, for example, between about 2 and about 10 (such as about any of 2, 3, 4, 5, 6, 7, 8, 9, or 10) amino acids in length may form the linkage between the transmembrane domain and the intracellular signaling domain of the anti-RTMC CAR. In some embodiments, the linker is a glycine- serine doublet.

[0235] In some embodiments, the transmembrane domain that naturally is associated with one of the sequences in the intracellular domain of the anti-RTMC CAR is used (e.g. , if an anti-RTMC

CAR intracellular domain comprises a CD28 co-stimulatory sequence, the transmembrane domain of the anti-RTMC CAR is derived from the CD28 transmembrane domain). In some embodiments, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to the transmembrane domains of the same or different surface membrane proteins to minimize interactions with other members of the receptor complex.

[0236] The intracellular signaling domain of the anti-RTMC CAR is responsible for activation of at least one of the normal effector functions of the immune cell in which the anti-RTMC CAR has been placed in. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines. Thus the term "intracellular signaling domain" refers to the portion of a protein which transduces the effector function signal and directs the cell to perform a specialized function. While usually the entire intracellular signaling domain can be employed, in many cases it is not necessary to use the entire chain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal. The term "intracellular signaling sequence" is thus meant to include any truncated portion of the intracellular signaling domain sufficient to transduce the effector function signal.

[0237] Examples of intracellular signaling domains for use in the anti-RTMC CAR of the invention include the cytoplasmic sequences of the T cell receptor (TCR) and co-receptors that act in concert to initiate signal transduction following antigen receptor engagement, as well as any derivative or variant of these sequences and any synthetic sequence that has the same functional capability.

[0238] It is known that signals generated through the TCR alone are insufficient for full activation of the T cell and that a secondary or co-stimulatory signal is also required. Thus, T cell activation can be said to be mediated by two distinct classes of intracellular signaling sequence: those that initiate antigen-dependent primary activation through the TCR (primary signaling sequences) and those that act in an antigen- independent manner to provide a secondary or co-stimulatory signal (co-stimulatory signaling sequences).

[0239] Primary signaling sequences regulate primary activation of the TCR complex either in a stimulatory way, or in an inhibitory way. Primary signaling sequences that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyro sine-based activation motifs or ITAMs. The anti-RTMC CAR constructs in some embodiments comprise one or more ITAMs.

[0240] Examples of IT AM containing primary signaling sequences that are of particular use in the invention include those derived from TCRC, FcRy, FcRp, CD3y, CD35, CD3s, CD5, CD22, CD79a, CD79b, and CD66d.

[0241] In some embodiments, the anti-RTMC CAR comprises a primary signaling sequence derived from CD3ζ. For example, the intracellular signaling domain of the CAR can comprise the CD3ζ intracellular signaling sequence by itself or combined with any other desired intracellular signaling sequence(s) useful in the context of the anti-RTMC CAR of the invention. For example, the intracellular domain of the anti-RTMC CAR can comprise a CD3ζ intracellular signaling sequence and a costimulatory signaling sequence. The costimulatory signaling sequence can be a portion of the intracellular domain of a costimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen- 1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like.

[0242] In some embodiments, the intracellular signaling domain of the anti-RTMC CAR comprises the intracellular signaling sequence of CD3ζ and the intracellular signaling sequence of CD28. In some embodiments, the intracellular signaling domain of the anti-RTMC CAR comprises the intracellular signaling sequence of CD3ζ and the intracellular signaling sequence of 4- IBB. In some embodiments, the intracellular signaling domain of the anti-RTMC CAR comprises the intracellular signaling sequence of CD3ζ and the intracellular signaling sequences of CD28 and 4- IBB.

[0243] Thus, for example, in some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, b) a transmembrane domain, and c) an intracellular signaling domain. In some embodiments, the

HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6),

HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT

181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-

A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the intracellular signaling domain is capable of activating an immune cell. In some

embodiments, the intracellular signaling domain comprises a primary signaling sequence and a co-stimulatory signaling sequence. In some embodiments, the primary signaling sequence comprises a CD3ζ intracellular signaling sequence. In some embodiments, the co- stimulatory signaling sequence comprises a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the intracellular domain comprises a CD3ζ intracellular signaling sequence and a

CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0244] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO:

6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, b) a transmembrane domain, and c) an intracellular signaling domain. In some embodiments, the intracellular signaling domain is capable of activating an immune cell. In some embodiments, the intracellular signaling domain comprises a primary signaling sequence and a co-stimulatory signaling sequence. In some embodiments, the primary signaling sequence comprises a CD3ζ intracellular signaling sequence. In some embodiments, the co-stimulatory signaling sequence comprises a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the intracellular domain comprises a CD3ζ intracellular signaling sequence and a CD28 and/or 4- 1BB intracellular signaling sequence.

[0245] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1,

2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-

249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID

NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or

3) amino acid substitutions, b) a transmembrane domain, and c) an intracellular signaling domain. In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID

NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID

NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253; b) an intracellular signaling domain. In some embodiments, the intracellular signaling domain is capable of activating an immune cell. In some embodiments, the intracellular signaling domain comprises a primary signaling sequence and a co-stimulatory signaling sequence. In some embodiments, the primary signaling sequence comprises a CD3ζ intracellular signaling sequence. In some embodiments, the co- stimulatory signaling sequence comprises a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the intracellular domain comprises a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0246] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of

1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; b) a transmembrane domain, and c) an intracellular signaling domain. In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1,

2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences; b) a transmembrane domain, and c) an intracellular signaling domain. In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; b) an intracellular signaling domain. In some embodiments, the intracellular signaling domain is capable of activating an immune cell. In some embodiments, the intracellular signaling domain comprises a primary signaling sequence and a co-stimulatory signaling sequence. In some embodiments, the primary signaling sequence comprises a CD3ζ intracellular signaling sequence. In some embodiments, the co-stimulatory signaling sequence comprises a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the intracellular domain comprises a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0247] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or

99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; b) a transmembrane domain, and c) an intracellular signaling domain. In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74; b) an intracellular signaling domain. In some embodiments, the intracellular signaling domain is capable of activating an immune cell. In some embodiments, the intracellular signaling domain comprises a primary signaling sequence and a co-stimulatory signaling sequence. In some embodiments, the primary signaling sequence comprises a CD3ζ intracellular signaling sequence. In some embodiments, the co- stimulatory signaling sequence comprises a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the intracellular domain comprises a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0248] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the

A02. In some embodiments, the MHC class I protein is HLA-A*02:01.

[0249] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO:

6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1

RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0250] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1,

2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247- 249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0251] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0252] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2,

3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of

1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-

189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0253] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0254] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0255] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%,

98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95%

(including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ

intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0256] In some embodiments, there is provided an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-

74; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0257] For example, in some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-

CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98,

126, 165, 191, and 209, respectively, SEQ ID NOs: 77, 99, 127, 164, 192, and 210, respectively,

SEQ ID NOs: 78, 100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167,

194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID

NOs: 81, 103, 131, 169, 191, and 214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and

215, respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82,

105, 134, 164, 192, and 217, respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218, respectively, SEQ ID NOs: 84, 107, 136, 172, 197, and 219, respectively, SEQ ID NOs: 85, 108,

140, 164, 192, and 208, respectively, SEQ ID NOs: 87, 111, 141, 175, 200, and 222, respectively, SEQ ID NOs: 85, 108, 142, 176, 192, and 208, respectively, SEQ ID NOs: 80, 112,

respectively, SEQ ID NOs: 82, 114, 145, 179, 202, and 225, respectively, SEQ ID NOs: 89, 115,

146, 175, 200, and 226, respectively, SEQ ID NOs: 90, 116, 147, 169, 191, and 227,

[0258] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

HC-CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ

ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and

209, respectively, SEQ ID NOs: 77, 99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78,

100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103,

respectively, SEQ ID NOs: 85, 108, 142, 176, 192, and 208, respectively, SEQ ID NOs: 80, 112, 143, 177, 191, and 223, respectively, SEQ ID NOs: 88, 113, 144, 178, 201, and 224,

respectively, SEQ ID NOs: 81, 117, 148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118, 149, 180, 199, and 228, respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229,

respectively, SEQ ID NOs: 91, 119, 151, 181, 191, and 230, respectively, SEQ ID NOs: 92, 120, 152, 182, 203, and 231, respectively, SEQ ID NOs: 80, 102, 153, 164, 192, and 232,

respectively, SEQ ID NOs: 93, 121, 154, 183, 204, and 233, respectively, SEQ ID NOs: 92, 120, 155, 184, 191, and 214, respectively, SEQ ID NOs: 80, 102, 156, 164, 192, and 234,

respectively, SEQ ID NOs: 85, 108, 157, 185, 200, and 235, respectively, SEQ ID NOs: 85, 108, 158, 186, 191, and 218, respectively, SEQ ID NOs: 79, 110, 159, 187, 205, and 236,

respectively, SEQ ID NOs: 92, 108, 160, 177, 191, and 218, respectively, SEQ ID NOs: 94, 122, 161, 173, 206, and 237, respectively, SEQ ID NOs: 95, 123, 162, 188, 200, and 238,

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively, or variants thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences and/or up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences.

[0259] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

143, 177, 191, and 223, respectively, SEQ ID NOs: 88, 113, 144, 178, 201, and 224, respectively, SEQ ID NOs: 82, 114, 145, 179, 202, and 225, respectively, SEQ ID NOs: 89, 115,

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively.

[0260] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of

SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ

ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and

56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively,

SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ

ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and

68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively,

SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively,

SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively.

[0261] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the anti-RTMC CAR comprises an anti- RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

Chimeric Antibody/T Cell Receptor

[0262] In some embodiments, the chimeric receptor is a chimeric antibody/T cell receptor construct (referred to herein as "abTCR"), and the anti-RTMC chimeric receptor is an anti- RTMC abTCR. In some embodiments, the anti-RTMC abTCR comprises a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module.

[0263] In some embodiments, the anti-RTMC abTCR comprises a first polypeptide chain and a second polypeptide chain. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage {e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the anti-RTMC abTCR is a heterodimer comprising the first polypeptide chain and the second polypeptide chain. In some embodiments, the first polypeptide chain and the second polypeptide chain are linked by at least one disulfide bond. The specificity of the anti-RTMC abTCR derives from an antibody moiety that confers binding specificity to the RTMC. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv. The capability of the anti-RTMC abTCR to recruit a TCR-associated signaling module derives from a T cell receptor module (TCRM). In some embodiments, the TCRM comprises the transmembrane module of a TCR (such as an aPTCR or a y5TCR). In some embodiments, the TCRM further comprises one or both of the connecting peptides or fragments thereof of a TCR. In some embodiments, the anti-RTMC abTCR further comprises at least one intracellular domain. In some embodiments, one or more of the at least one intracellular domain of the anti-RTMC abTCR comprises a sequence from the intracellular domain of a TCR. In some embodiments, one or more of the at least one

intracellular domain of the anti-RTMC abTCR comprises a T cell costimulatory signaling sequence. The costimulatory signaling sequence can be a portion of the intracellular domain of a costimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen- 1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like. In some embodiments, the antibody moiety is contained in an extracellular domain of the anti-RTMC abTCR. In some embodiments, the anti-RTMC abTCR further comprises one or more peptide linkers between the antibody moiety and the TCRM to optimize the length of the extracellular domain.

[0264] In some embodiments, the antibody moiety is a Fab-like antigen-binding module comprising a) a first polypeptide chain comprising a first antigen-binding domain comprising a V_H antibody domain and a C_HI antibody domain and b) a second polypeptide chain comprising a second antigen-binding domain comprising a V_L antibody domain and a C_L antibody domain. In some embodiments, the first antigen-binding domain comprises the V_H antibody domain amino- terminal to the C_HI antibody domain and/or the second antigen-binding domain comprises the V_L antibody domain amino-terminal to the C_L antibody domain. In some embodiments, there is a peptide linker between the V_L and C_L antibody domains and/or a peptide linker between the V_H and C_HI antibody domains. In some embodiments, all of the V_L antibody domain and V_H antibody domain CDRs are derived from the same antibody moiety. In some embodiments, the V_L antibody domain and the V_H antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g. , peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second antigen-binding domains are linked by a disulfide bond. In some embodiments, the first and second antigen-binding domains are linked by a disulfide bond between a residue in the CHI domain and a residue in the CL domain. In some embodiments, the CHI domain is derived from an IgG (e.g, IgGl, IgG2, IgG3, or IgG4) heavy chain, optionally human. In some embodiments, the CHI domain is a variant comprising one or more modifications (e.g. , amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the CL domain is derived from a kappa or lambda light chain, optionally human. In some embodiments, the CL domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the CHI and/or CL domains comprise one or more modifications that do not substantially alter their binding affinities for one another. In some embodiments, the CHI and/or CL domains comprise one or more modifications that increase their binding affinities for one another and/or introduce a non-naturally occurring disulfide bond. In some embodiments, the Fab-like antigen-binding module is human, humanized, chimeric, semi-synthetic, or fully synthetic.

[0265] In some embodiments, the antibody moiety is a Fab-like antigen-binding module comprising a) a first polypeptide chain comprising a first antigen-binding domain comprising a

VL antibody domain and a CHI antibody domain and b) a second polypeptide chain comprising a second antigen-binding domain comprising a VH antibody domain and a CL antibody domain. In some embodiments, the first antigen-binding domain comprises the VL antibody domain amino- terminal to the CHI antibody domain and/or the second antigen-binding domain comprises the

VH antibody domain amino-terminal to the CL antibody domain. In some embodiments, there is a peptide linker between the VH and CL antibody domains and/or a peptide linker between the VL and CHI antibody domains. In some embodiments, all of the VL antibody domain and VH antibody domain CDRs are derived from the same antibody moiety. In some embodiments, the

VL antibody domain and the VH antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g. , peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second antigen-binding domains are linked by a disulfide bond. In some embodiments, the first and second antigen-binding domains are linked by a disulfide bond between a residue in the CHI domain and a residue in the CL domain. In some embodiments, the CHI domain is derived from an IgG (e.g, IgGl, IgG2, IgG3, or IgG4) heavy chain, optionally human. In some embodiments, the CHI domain is a variant comprising one or more modifications (e.g. , amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the CL domain is derived from a kappa or lambda light chain, optionally human. In some embodiments, the CL domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the CHI and/or CL domains comprise one or more modifications that do not substantially alter their binding affinities for one another. In some embodiments, the CHI and/or CL domains comprise one or more modifications that increase their binding affinities for one another and/or introduce a non-naturally occurring disulfide bond. In some embodiments, the Fab-like antigen-binding module is human, humanized, chimeric, semi-synthetic, or fully synthetic.

[0266] In some embodiments, the antibody moiety is an Fv-like antigen-binding module comprising a) a first polypeptide chain comprising a first antigen-binding domain comprising a

VH antibody domain and optionally a first TCR constant domain from a T cell receptor subunit; and b) a second polypeptide chain comprising a second antigen-binding domain comprising a VL antibody domain and optionally a second TCR constant domain from a T cell receptor subunit.

In some embodiments, the first antigen-binding domain comprises the VH antibody domain amino-terminal to the first TCR constant domain and/or the second antigen-binding domain comprises the VL antibody domain amino-terminal to the second TCR constant domain. In some embodiments, there is a peptide linker between the VL antibody domain and the first TCR constant domain and/or a peptide linker between the VH antibody domain and the second TCR constant domain. In some embodiments, all of the VL antibody domain and VH antibody domain

CDRs are derived from the same antibody moiety. In some embodiments, the VL antibody domain and the VH antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g. , peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second antigen-binding domains are linked by a disulfide bond.

In some embodiments, the first and second antigen-binding domains are linked by a disulfide bond between a residue in the first TCR constant domain and a residue in the second TCR constant domain. In some embodiments, the first TCR constant domain is derived from a TCR a subunit, optionally human, and/or the second TCR constant domain is derived from a TCR β subunit, optionally human. In some embodiments, the first TCR constant domain is derived from a TCR a subunit comprising the amino acid sequence of SEQ ID NO: 281, and/or the second TCR constant domain is derived from a TCR β subunit, comprising the amino acid sequence of SEQ ID NO: 282. In some embodiments, the first TCR constant domain is derived from a TCR δ subunit, optionally human, and/or the second TCR constant domain is derived from a TCR γ subunit, optionally human. In some embodiments, the first TCR constant domain is derived from a TCR δ subunit comprising the amino acid sequence of SEQ ID NO: 283, and/or the second TCR constant domain is derived from a TCR γ subunit, comprising the amino acid sequence of SEQ ID NO: 282. In some embodiments, the first and/or second TCR constant domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the first and/or second TCR constant domains comprise one or more modifications that do not substantially alter their binding affinities for one another. In some embodiments, the first and/or second TCR constant domains comprise one or more modifications that increase their binding affinities for one another and/or introduce a non-naturally occurring disulfide bond. In some embodiments, the Fv-lrke antigen-binding module is human, humanized, chimeric, semisynthetic, or fully synthetic.

[0267] In some embodiments, the antibody moiety is an scFv comprising a) a polypeptide chain comprising a VH antibody domain and a VL antibody domain. In some embodiments, the scFv comprises the VH antibody domain amino-terminal to the VL antibody domain. In some embodiments, the scFv comprises the VL antibody domain amino-terminal to the VH antibody domain. In some embodiments, there is a peptide linker between the VL antibody domain and the VH antibody domain. In some embodiments, all of the VL antibody domain and VH antibody domain CDRs are derived from the same antibody moiety. In some embodiments, the VL antibody domain and the VH antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the scFv is human, humanized, chimeric, semisynthetic, or fully synthetic.

[0268] In some embodiments, the TCRM comprises a) a first polypeptide chain comprising a first T cell receptor domain (TCRD) comprising a first transmembrane domain and b) a second polypeptide chain comprising a second TCRD comprising a second transmembrane domain. In some embodiments, the first transmembrane domain is the transmembrane domain of a first

TCR subunit and/or the second transmembrane domain is the transmembrane domain of a second TCR subunit. In some embodiments, the first TCR subunit is a TCR a chain (e.g. ,

GenBank Accession No: CCI73895), and the second TCR subunit is a TCR β chain (e.g. ,

GenBank Accession No: CCI73893). In some embodiments, the first TCR subunit is a TCR β chain, and the second TCR subunit is a TCR a chain. In some embodiments, the first TCR subunit is a TCR γ chain (e.g. , GenBank Accession No: AGE91788), and the second TCR subunit is a TCR δ chain (e.g. , GenBank Accession No: AAQ57272). In some embodiments, the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain. In some embodiments, the first and/or second transmembrane domains comprise (such as consist of), individually, a transmembrane domain contained in any one of the TCR subunit amino acid sequences of SEQ ID NOs: 281-284. In some embodiments, the first and/or second

transmembrane domains comprise (such as consist of), individually, any one of the amino acid sequences of SEQ ID NOs: 285-288. In some embodiments, the first TCRD further comprises a first connecting peptide amino-terminal to the transmembrane domain and/or the second TCRD further comprises a second connecting peptide amino-terminal to the transmembrane domain. In some embodiments, the first connecting peptide comprises all or a portion of the connecting peptide of the first TCR subunit and/or the second connecting peptide comprises all or a portion of the connecting peptide of the second TCR subunit. In some embodiments, the first and/or second connecting peptides comprise (such as consist of), individually, all or a portion of a connecting peptide contained in any one of the TCR subunit amino acid sequences of SEQ ID

NOs: 281-284. In some embodiments, the first and/or second connecting peptides comprise

(such as consist of), individually, any one of the amino acid sequences of SEQ ID NOs: 289-

296. In some embodiments, the first TCRD further comprises a first TCR intracellular domain carboxy-terminal to the first transmembrane domain and/or the second TCRD further comprises a second TCR intracellular domain carboxy-terminal to the second transmembrane domain. In some embodiments, the first TCR intracellular domain comprises all or a portion of the intracellular domain of the first TCR subunit and/or the second TCR intracellular domain comprises all or a portion of the intracellular domain of the second TCR subunit. In some embodiments, the first and/or second TCR intracellular domains comprise (such as consist of), individually, all or a portion of an intracellular sequence contained in any one of the TCR subunit amino acid sequences of SEQ ID NOs: 281-284. In some embodiments, the first and/or second TCR intracellular domains comprise (such as consist of), individually, any one of the amino acid sequences of SEQ ID NOs: 297-298. In some embodiments, the first TCRD is a fragment of the first TCR subunit and/or the second TCRD is a fragment of the second TCR chain. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g. , peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second TCRDs are linked by a disulfide bond. In some embodiments, the first and second TCRDs are linked by a disulfide bond between a residue in the first connecting peptide and a residue in the second connecting peptide. In some embodiments, the TCRM is capable of recruiting at least one TCR-associated signaling module selected from the group consisting of CD35s, CD3y8, and ζζ. In some embodiments, the TCRM is capable of recruiting each of CD35s, CD3y8, and ζζ to form an octameric anti-RTMC abTCR-CD3 complex (i.e., promotes anti-RTMC abTCR-CD3 complex formation).

[0269] In some embodiments, the anti-RTMC abTCR is a molecule comprising a fusion of the antibody moiety to the TCRM. In some embodiments, the anti-RTMC abTCR comprises a fusion of the first polypeptide chain of the Fab-like or Fv-like antigen-binding module amino- terminal to the first polypeptide chain of the TCRM, thereby forming a first polypeptide chain of the anti-RTMC abTCR, and a fusion of the second polypeptide chain of the Fab-like or Fv-like antigen-binding module amino-terminal to the second polypeptide chain of the TCRM, thereby forming a second polypeptide chain of the anti-RTMC abTCR. In some embodiments, the anti- RTMC abTCR comprises a fusion of the scFv amino-terminal to the first or second polypeptide chain of the TCRM. In some embodiments, the anti-RTMC abTCR further comprises a peptide linker between the first polypeptide chain of the Fab-like or Fv-like antigen-binding module and the first polypeptide chain of the TCRM and/or a peptide linker between the second polypeptide chain of the Fab-like or Fv-like antigen-binding module and the second polypeptide chain of the TCRM. In some embodiments, the anti-RTMC abTCR further comprises a peptide linker between the scFv and the first or second polypeptide chain of the TCRM. In some embodiments, the peptide linker is between about 5 to about 70 (such as about any of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70, including any ranges between these values) amino acids in length. In some embodiments, the first polypeptide chain of the anti-RTMC abTCR further comprises an amino-terminal first signal peptide and/or the second polypeptide chain of the anti-RTMC abTCR further comprises an amino-terminal second signal peptide. In some embodiments, the first and/or second signal peptides comprise (such as consist of) the amino acid sequence of SEQ ID NO: 299. In some embodiments, the first polypeptide chain of the anti-RTMC abTCR further comprises a first accessory intracellular domain carboxy-terminal to the first transmembrane domain and/or the second polypeptide chain of the anti-RTMC abTCR further comprises a second accessory intracellular domain carboxy-terminal to the second transmembrane domain. In some embodiments, the first and/or second accessory intracellular domains comprise a TCR costimulatory domain. In some embodiments, the TCR costimulatory domain comprises all or a portion of the amino acid sequence of SEQ ID NO: 300. In some embodiments, the first and second polypeptide chains of the anti-RTMC abTCR are linked, such as by a covalent linkage (e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the anti-RTMC abTCR is a heterodimer.

[0270] Thus, for example, in some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the HIV-1 RT peptide is HIV- 1 RT 181 (SEQ ID NO:

5) , HIV- 1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV- 1 RT 181 Y181C (SEQ ID NO: 8), or HIV- 1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv. In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV- 1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV- 1 RT peptide and a different subtype of the MHC class I protein.

[0271] In some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV- 1 RT 181 (SEQ ID NO: 5), HIV- 1 RT 181 M184V (SEQ ID NO:

6) , HIV- 1 RT 181 M184I (SEQ ID NO: 7), HIV- 1 RT 181 Y181C (SEQ ID NO: 8), or HIV- 1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv.

[0272] In some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247- 249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, there is provided an anti- RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247- 249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250- 253; and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv.

[0273] In some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2,

189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID

NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of

SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2,

3, 4, or 5) amino acid substitutions; and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-

RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-

CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-

CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-

CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the

LC-CDR sequences; and b) a T cell receptor module (TCRM) capable of recruiting at least one

TCR-associated signaling module. In some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-

189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv.

[0274] In some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, there is provided an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74; and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv.

[0275] For example, in some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-

respectively, SEQ ID NOs: 80, 102, 139, 174, 199, and 221, respectively, SEQ ID NOs: 79, 110, 140, 164, 192, and 208, respectively, SEQ ID NOs: 87, 111, 141, 175, 200, and 222,

[0276] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

140, 164, 192, and 208, respectively, SEQ ID NOs: 87, 111, 141, 175, 200, and 222, respectively, SEQ ID NOs: 85, 108, 142, 176, 192, and 208, respectively, SEQ ID NOs: 80, 112, 143, 177, 191, and 223, respectively, SEQ ID NOs: 88, 113, 144, 178, 201, and 224,

[0277] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively.

[0278] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of

SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively,

[0279] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the anti-RTMC abTCR comprises an anti- RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

[0280] Also provided herein are effector cells (such as lymphocytes, e.g., T cells) expressing an anti-RTMC chimeric receptor, such as an anti-RTMC CAR or anti-RTMC abTCR.

[0281] Also provided is a method of producing an effector cell expressing an anti-RTMC CAR or anti-RTMC abTCR, the method comprising introducing a vector comprising a nucleic acid encoding the anti-RTMC CAR or anti-RTMC abTCR into the effector cell. In some

embodiments, introducing the vector into the effector cell comprises transducing the effector cell with the vector. In some embodiments, introducing the vector into the effector cell comprises transfecting the effector cell with the vector. Transduction or transfection of the vector into the effector cell can be carried about using any method known in the art. Immunoconj ugates

[0282] The anti-RTMC constructs in some embodiments comprise an immunoconjugate comprising an anti-RTMC antibody moiety attached to an effector molecule (also referred to herein as an "anti-RTMC immunoconjugate"). In some embodiments the effector molecule is a therapeutic agent, such as a viral therapeutic agent, which is either cytotoxic, cytostatic or otherwise provides some therapeutic benefit. In some embodiments, the effector molecule is a label, which can generate a detectable signal, either directly or indirectly.

[0283] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising an anti-RTMC antibody moiety and a therapeutic agent (also referred to herein as an "antibody- drug conjugate", or "ADC"). In some embodiments, the therapeutic agent is a toxin that is either cytotoxic, cytostatic or otherwise prevents or reduces the ability of the target cells to divide. The use of ADCs for the local delivery of cytotoxic or cytostatic agents, i.e., drugs to kill or inhibit tumor cells in the treatment of cancer (Syrigos and Epenetos, Anticancer Research 19:605-614 (1999); Niculescu-Duvaz and Springer, Adv. Org. Del. Rev. 26: 151 -172 (1997); U.S. Patent No. 4,975,278) allows targeted delivery of the drug moiety to target cells, and intracellular accumulation therein, where systemic administration of these unconjugated therapeutic agents may result in unacceptable levels of toxicity to normal cells as well as the target cells sought to be eliminated (Baldwin et al, Lancet (Mar. 15, 1986):603-605 (1986); Thorpe, (1985)

"Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review," in Monoclonal Antibodies '84: Biological And Clinical Applications, A. Pinchera et al. (eds.), pp. 475- 506). Maximal efficacy with minimal toxicity is sought thereby. Importantly, since most normal cells do not present the RTMC on their surface, they cannot bind the anti-RTMC immunoconjugate, and are protected from the killing effect of the toxin or other therapeutic agents.

[0284] Therapeutic agents used in anti-RTMC immunoconjugates include, for example, daunomycin, doxorubicin, methotrexate, and vindesine (Rowland et al, Cancer Immunol.

Immunother. 21: 183-187 (1986)). Toxins used in anti-RTMC immunoconjugates include bacterial toxins such as diphtheria toxin, plant toxins such as ricin, small molecule toxins such as geldanamycin (Mandler et al, J.Nat. Cancer Inst. 92(19): 1573-1581 (2000); Mandler et al, Bioorganic & Med. Chem. Letters 10: 1025- 1028 (2000); Mandler et al, Bioconjugate Chem. 13:786-791 (2002)), maytansinoids (EP 1391213; Liu et al, Proc. Natl. Acad. Sci. USA

93:8618-8623 (1996)), and calicheamicin (Lode et al, Cancer Res. 58:2928 (1998); Hinman et al, Cancer Res. 53:3336-3342 (1993)). The toxins may exert their cytotoxic and cytostatic effects by mechanisms including tubulin binding, DNA binding, or topoisomerase inhibition. Some cytotoxic drugs tend to be inactive or less active when conjugated to large antibodies or protein receptor ligands.

[0285] Enzymatically active toxins and fragments thereof that can be used include, for example, diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain,a-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. See, e.g. , WO 93/21232 published October 28, 1993.

[0286] Anti-RTMC immunoconjugates of an anti-RTMC antibody moiety and one or more small molecule toxins, such as a calicheamicin, maytansinoids, dolastatins, aurostatins, a trichothecene, and CC1065, and the derivatives of these toxins that have toxin activity, are also contemplated herein.

[0287] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a therapeutic agent that has an intracellular activity. In some embodiments, the anti-RTMC immunoconjugate is internalized and the therapeutic agent is a cytotoxin that blocks the protein synthesis of the cell, therein leading to cell death. In some embodiments, the therapeutic agent is a cytotoxin comprising a polypeptide having ribosome-inactivating activity including, for example, gelonin, bouganin, saporin, ricin, ricin A chain, bryodin, diphtheria toxin, restrictocin, Pseudomonas exotoxin A and variants thereof. In some embodiments, where the therapeutic agent is a cytotoxin comprising a polypeptide having a ribosome-inactivating activity, the anti- RTMC immunoconjugate must be internalized upon binding to the target cell in order for the protein to be cytotoxic to the cells.

[0288] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a therapeutic agent that acts to disrupt DNA. In some embodiments, the therapeutic agent that acts to disrupt DNA is, for example, selected from the group consisting of enediyne (e.g. , calicheamicin and esperamicin) and non-enediyne small molecule agents (e.g., bleomycin, methidiumpropyl-EDTA-Fe(II)).

[0289] The present invention further contemplates an anti-RTMC immunoconjugate formed between the anti-RTMC antibody moiety and a compound with nucleo lytic activity (e.g. , a ribonuclease or a DNA endonuclease such as a deoxyribonuclease; DNase). [0290] In some embodiments, the anti-RTMC immunoconjugate comprises an agent that acts to disrupt tubulin. Such agents may include, for example, rhizoxin/maytansine, paclitaxel, vincristine and vinblastine, colchicine, auristatin dolastatin 10 MMAE, and peloruside A.

[0291] In some embodiments, the anti-RTMC immunoconjugate comprises an alkylating agent including, for example, Asaley NSC 167780, AZQ NSC 182986, BCNU NSC 409962, Busulfan NSC 750, carboxyphthalatoplatinum NSC 271674, CBDCA NSC 241240, CCNU NSC 79037, CHIP NSC 256927, chlorambucil NSC 3088, chlorozotocin NSC 178248, cis-platinum NSC 119875, clomesone NSC 338947, cyanomorpholinodoxorubicin NSC 357704, cyclodisone NSC 348948, dianhydrogalactitol NSC 132313, fluorodopan NSC 73754, hepsulfam NSC 329680, hycanthone NSC 142982, melphalan NSC 8806, methyl CCNU NSC 95441 , mitomycin C NSC 26980, mitozolamide NSC 353451 , nitrogen mustard NSC 762, PCNU NSC 95466, piperazine NSC 344007, piperazinedione NSC 135758, pipobroman NSC 25154, porfiromycin NSC 56410, spirohydantoin mustard NSC 172112, teroxirone NSC 296934, tetraplatin NSC 363812, thio- tepa NSC 6396, triethylenemelamine NSC 9706, uracil nitrogen mustard NSC 34462, and Yoshi-864 NSC 102627.

[0292] In some embodiments, the anti-RTMC immunoconjugate comprises a highly radioactive atom. A variety of radioactive isotopes are available for the production of radioconjugated antibodies. Examples include ²¹¹At, ¹³¹I, ¹²⁵I, ⁹⁰Y, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁵³Sm, ²¹²Bi, ³²P, ²¹²Pb and radioactive isotopes of Lu.

[0293] In some embodiments, the anti-RTMC antibody moiety can be conjugated to a "receptor" (such as streptavidin) for utilization in tumor pre-targeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a "ligand" (e.g., avidin) that is conjugated to a cytotoxic agent (e.g., a radio nucleotide).

[0294] In some embodiments, an anti-RTMC immunoconjugate may comprise an anti-RTMC antibody moiety conjugated to a prodrug-activating enzyme. In some such embodiments, a prodrug-activating enzyme converts a prodrug to an active drug, such as an anti- viral drug. Such anti-RTMC immunoconjugates are useful, in some embodiments, in antibody-dependent enzyme- mediated prodrug therapy ("ADEPT"). Enzymes that may be conjugated to an antibody include, but are not limited to, alkaline phosphatases, which are useful for converting phosphate- containing prodrugs into free drugs; arylsulfatases, which are useful for converting sulfate- containing prodrugs into free drugs; proteases, such as serratia protease, thermolysin, subtilisin, carboxypeptidases and cathepsins (such as cathepsins B and L), which are useful for converting peptide-containing prodrugs into free drugs; D-alanylcarboxypeptidases, which are useful for converting prodrugs that contain D-amino acid substituents; carbohydrate-cleaving enzymes such as β-galactosidase and neuraminidase, which are useful for converting glycosylated prodrugs into free drugs; β-lactamase, which is useful for converting drugs derivatized with β - lactams into free drugs; and penicillin amidases, such as penicillin V amidase and penicillin G amidase, which are useful for converting drugs derivatized at their amine nitrogens with phenoxyacetyl or phenylacetyl groups, respectively, into free drugs. In some embodiments, enzymes may be covalently bound to antibody moieties by recombinant DNA techniques well known in the art. See, e.g., Neuberger et al, Nature 312:604-608 (1984).

[0295] In some embodiments, the therapeutic portion of the anti-RTMC immunoconjugates may be a nucleic acid. Nucleic acids that may be used include, but are not limited to, anti-sense RNA, genes or other polynucleotides, including nucleic acid analogs such as thioguanine and thiopurine.

[0296] The present application further provides anti-RTMC immunoconjugates comprising an anti-RTMC antibody moiety attached to an effector molecule, wherein the effector molecule is a label, which can generate a detectable signal, indirectly or directly. These anti-RTMC

immunoconjugates can be used for research or diagnostic applications, such as for the in vivo detection of cancer. The label is preferably capable of producing, either directly or indirectly, a detectable signal. For example, the label may be radio-opaque or a radioisotope, such as 3 H, 14 C,

32 P, 35 S, 123 I, 125 I, 131 I; a fluorescent (fluorophore) or chemiluminescent (chromophore) compound, such as fluorescein isothiocyanate, rhodamine or luciferin; an enzyme, such as alkaline phosphatase,P-galactosidase or horseradish peroxidase; an imaging agent; or a metal ion. In some embodiments, the label is a radioactive atom for scintigraphic studies, for example

9 "9Tc or or a spin label for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI), such as zirconium-89, iodine-123, iodine-131, indium-I l l, fluorine- 19, carbon- 13, nitrogen- 15, oxygen- 17, gadolinium, manganese or iron. Zirconium-89 may be complexed to various metal chelating agents and conjugated to antibodies, e.g., for PET imaging (WO 2011/056983).

[0297] In some embodiments, the anti-RTMC immunoconjugate is detectable indirectly. For example, a secondary antibody that is specific for the anti-RTMC immunoconjugate and contains a detectable label can be used to detect the anti-RTMC immunoconjugate.

[0298] Thus, for example, in some embodiments, there is provided an anti-RTMC

immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, and b) an effector molecule. In some embodiments, the HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV- 1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the effector molecule is covalently attached to the anti-RTMC antibody moiety. In some embodiments, the effector molecule is a therapeutic agent selected, for example, from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid. In some embodiments, the effector molecular is a viral therapeutic agent. In some embodiments, the viral therapeutic agent is a highly radioactive atom selected, for example, from the group consisting of ²¹¹At, ¹³¹I, ¹²⁵I, ⁹⁰Y, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁵³Sm,

In some embodiments, the effector molecule is a label that can generate a detectable signal, either directly or indirectly. In some embodiments, the label is a radioisotope selected, for example, from the group consisting of 3 H, 14 C, 32 P, 35 S, 123 I, 125 I, and 131 I. In some embodiments, the anti-RTMC antibody moiety is an scFv. In some embodiments, the anti- RTMC antibody moiety is human, humanized, or semi- synthetic. In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, 5, or 6) complex comprising the MHC class I protein and a variant of the HIV-1 RT peptide having one amino acid substitution (such as a conservative amino acid substitution). In some embodiments, the anti-RTMC antibody moiety cross-reacts with at least one (such as at least any of 2, 3, 4, or 5) complex comprising the HIV-1 RT peptide and a different subtype of the MHC class I protein.

[0299] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT

181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID

NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID

NO: 9) peptide and HLA-A*02:01, and b) an effector molecule. In some embodiments, the effector molecule is covalently attached to the anti-RTMC antibody moiety. In some

embodiments, the effector molecule is a therapeutic agent selected, for example, from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid. In some embodiments, the effector molecular is a viral therapeutic agent. In some embodiments, the viral therapeutic agent is a highly radioactive atom selected, for example, from the group consisting of ²¹¹At, ¹³¹I, ¹²⁵I, ⁹⁰Y, ¹⁸⁶Re, ¹⁸⁸Re, ¹⁵³Sm, ²¹²Bi, ³²P, and ²¹²Pb. In some embodiments, the effector molecule is a label that can generate a detectable signal, either directly or indirectly. In some embodiments, the label is a radioisotope selected, for example, from the group consisting of ³H, ¹⁴C, ³²P, ³⁵S, ¹²³I, ¹²⁵I, and ¹³¹I. In some embodiments, the anti-RTMC antibody moiety is an scFv. In some embodiments, the anti-RTMC antibody moiety is human, humanized, or semisynthetic.

[0300] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and b) an effector molecule.

[0301] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, an HC- CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC- CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, and b) an effector molecule.

[0302] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an

HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, an LC- CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 (such as about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions, and b) an effector molecule.

[0303] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75- 96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences, and b) an effector molecule.

[0304] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-

96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC-CDR 1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239, and b) an effector molecule.

[0305] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47- 74, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and b) an effector molecule.

[0306] In some embodiments, there is provided an anti-RTMC immunoconjugate comprising a) an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, b) an effector molecule.

[0307] For example, in some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-

CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164,

190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively, SEQ ID

NOs: 77, 99, 127, 164, 192, and 210, respectively, SEQ ID NOs: 78, 100, 128, 166, 193, and

211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80,

102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and 214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76, 98,

145, 179, 202, and 225, respectively, SEQ ID NOs: 89, 115, 146, 175, 200, and 226, respectively, SEQ ID NOs: 90, 116, 147, 169, 191, and 227, respectively, SEQ ID NOs: 81, 117, 148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118, 149, 180, 199, and 228,

[0308] In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-

146, 175, 200, and 226, respectively, SEQ ID NOs: 90, 116, 147, 169, 191, and 227, respectively, SEQ ID NOs: 81, 117, 148, 169, 191, and 218, respectively, SEQ ID NOs: 82, 118, 149, 180, 199, and 228, respectively, SEQ ID NOs: 82, 114, 150, 176, 200, and 229,

[0309] In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively.

[0310] In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively,

SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ

ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and

58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively,

SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ

ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and

70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively,

SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%,

98%, or 99%) sequence identity. In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID

NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ

ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and

55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively,

SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively.

[0311] In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC immunoconjugate comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

Nucleic Acids

[0312] Nucleic acid molecules encoding the anti-RTMC constructs or anti-RTMC antibody moieties are also contemplated. In some embodiments, there is provided a nucleic acid (or a set of nucleic acids) encoding a full-length anti-RTMC antibody. In some embodiments, there is provided a nucleic acid (or a set of nucleic acids) encoding a multi- specific anti-RTMC molecule (e.g. , a multi- specific anti-RTMC antibody, a bispecific anti-RTMC antibody, or a bispecific T-cell engager anti-RTMC antibody), or polypeptide portion thereof. In some embodiments, there is provided a nucleic acid (or a set of nucleic acids) encoding an anti-RTMC CAR or anti-RTMC abTCR. In some embodiments, there is provided a nucleic acid (or a set of nucleic acids) encoding an anti-RTMC immunoconjugate, or polypeptide portion thereof. [0313] The present application also includes variants to these nucleic acid sequences. For example, the variants include nucleotide sequences that hybridize to the nucleic acid sequences encoding the anti-RTMC constructs or anti-RTMC antibody moieties of the present application under at least moderately stringent hybridization conditions.

[0314] The present invention also provides vectors in which a nucleic acid of the present invention is inserted.

[0315] In brief summary, the expression of an anti-RTMC construct (e.g. , anti-RTMC CAR or anti-RTMC abTCR) or polypeptide portion thereof by a natural or synthetic nucleic acid encoding the anti-RTMC construct or polypeptide portion thereof can be achieved by inserting the nucleic acid into an appropriate expression vector, such that the nucleic acid is operably linked to 5' and 3' regulatory elements, including for example a promoter (e.g. , a lymphocyte- specific promoter) and a 3' untranslated region (UTR). The vectors can be suitable for replication and integration in eukaryotic host cells. Typical cloning and expression vectors contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence.

[0316] The nucleic acids of the present invention may also be used for nucleic acid

immunization and gene therapy, using standard gene delivery protocols. Methods for gene delivery are known in the art. See, e.g. , U.S. Pat. Nos. 5,399,346, 5,580,859, 5,589,466, incorporated by reference herein in their entireties. In some embodiments, the invention provides a gene therapy vector.

[0317] The nucleic acid can be cloned into a number of types of vectors. For example, the nucleic acid can be cloned into a vector including, but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.

[0318] Further, the expression vector may be provided to a cell in the form of a viral vector. Viral vector technology is well known in the art and is described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and in other virology and molecular biology manuals. Viruses which are useful as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers (see, e.g. , WO 01/96584; WO 01/29058; and U.S. Pat. No.

6,326, 193). [0319] A number of viral based systems have been developed for gene transfer into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. A selected gene can be inserted into a vector and packaged in retroviral particles using techniques known in the art. The recombinant virus can then be isolated and delivered to cells of the subject either in vivo or ex vivo. A number of retroviral systems are known in the art. In some

embodiments, adenovirus vectors are used. A number of adenovirus vectors are known in the art. In some embodiments, lentivirus vectors are used. Vectors derived from retroviruses such as the lentivirus are suitable tools to achieve long-term gene transfer since they allow long-term, stable integration of a transgene and its propagation in daughter cells. Lentiviral vectors have the added advantage over vectors derived from onco-retroviruses such as murine leukemia viruses in that they can transduce non-proliferating cells, such as hepatocytes. They also have the added advantage of low immunogenicity.

[0320] Additional promoter elements, e.g., enhancers, regulate the frequency of transcriptional initiation. Typically, these are located in the region 30-110 bp upstream of the start site, although a number of promoters have recently been shown to contain functional elements downstream of the start site as well. The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or moved relative to one another. In the thymidine kinase (tk) promoter, the spacing between promoter elements can be increased to 50 bp apart before activity begins to decline.

[0321] One example of a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence capable of driving high levels of expression of any polynucleotide sequence operatively linked thereto.

Another example of a suitable promoter is Elongation Growth Factor- la (EF-la). However, other constitutive promoter sequences may also be used, including, but not limited to the simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human

immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, an avian leukemia virus promoter, an Epstein-Barr virus immediate early promoter, a Rous sarcoma virus promoter, as well as human gene promoters such as, but not limited to, the actin promoter, the myosin promoter, the hemoglobin promoter, and the creatine kinase promoter. Further, the invention should not be limited to the use of constitutive promoters. Inducible promoters are also contemplated as part of the invention. The use of an inducible promoter provides a molecular switch capable of turning on expression of the polynucleotide sequence which it is operatively linked when such expression is desired, or turning off the expression when expression is not desired. Examples of inducible promoters include, but are not limited to a metallothionine promoter, a glucocorticoid promoter, a progesterone promoter, and a tetracycline promoter.

[0322] In order to assess the expression of a polypeptide or portions thereof, the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of expressing cells from the population of cells sought to be transfected or infected through viral vectors. In other aspects, the selectable marker may be carried on a separate piece of DNA and used in a co-transfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers include, for example, antibiotic- resistance genes, such as neo and the like.

[0323] Reporter genes are used for identifying potentially transfected cells and for evaluating the functionality of regulatory sequences. In general, a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and that encodes a polypeptide whose expression is manifested by some easily detectable property, e.g., enzymatic activity. Expression of the reporter gene is assayed at a suitable time after the DNA has been introduced into the recipient cells. Suitable reporter genes may include genes encoding luciferase, β-galactosidase, chloramphenicol acetyl transferase, secreted alkaline phosphatase, or the green fluorescent protein gene (e.g., Ui-Tel et al., 2000 FEBS Letters 479: 79-82). Suitable expression systems are well known and may be prepared using known techniques or obtained commercially. In general, the construct with the minimal 5' flanking region showing the highest level of expression of reporter gene is identified as the promoter. Such promoter regions may be linked to a reporter gene and used to evaluate agents for the ability to modulate promoter-driven transcription.

[0324] Methods of introducing and expressing genes into a cell are known in the art. In the context of an expression vector, the vector can be readily introduced into a host cell, e.g., mammalian, bacterial, yeast, or insect cell by any method in the art. For example, the expression vector can be transferred into a host cell by physical, chemical, or biological means.

[0325] Physical methods for introducing a polynucleotide into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. Methods for producing cells comprising vectors and/or exogenous nucleic acids are well-known in the art. See, for example, Sambrook et al. (2001, Molecular Cloning: A

Laboratory Manual, Cold Spring Harbor Laboratory, New York). In some embodiments, the introduction of a polynucleotide into a host cell is carried out by calcium phosphate transfection. [0326] Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells. Other viral vectors can be derived from lentivirus, poxviruses, herpes simplex virus 1, adenoviruses and adeno-associated viruses, and the like. See, for example, U.S. Pat. Nos. 5,350,674 and

5,585,362.

[0327] Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as a delivery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle).

[0328] In the case where a non- viral delivery system is utilized, an exemplary delivery vehicle is a liposome. The use of lipid formulations is contemplated for the introduction of the nucleic acids into a host cell (in vitro, ex vivo or in vivo). In another aspect, the nucleic acid may be associated with a lipid. The nucleic acid associated with a lipid may be encapsulated in the aqueous interior of a liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via a linking molecule that is associated with both the liposome and the

oligonucleotide, entrapped in a liposome, complexed with a liposome, dispersed in a solution containing a lipid, mixed with a lipid, combined with a lipid, contained as a suspension in a lipid, contained or complexed with a micelle, or otherwise associated with a lipid. Lipid, lipid/DNA or lipid/expression vector associated compositions are not limited to any particular structure in solution. For example, they may be present in a bilayer structure, as micelles, or with a "collapsed" structure. They may also simply be interspersed in a solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty substances which may be naturally occurring or synthetic lipids. For example, lipids include the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds which contain long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes.

[0329] Regardless of the method used to introduce exogenous nucleic acids into a host cell or otherwise expose a cell to the inhibitor of the present invention, in order to confirm the presence of the recombinant DNA sequence in the host cell, a variety of assays may be performed. Such assays include, for example, "molecular biological" assays well known to those of skill in the art, such as Southern and Northern blotting, RT-PCR and PCR; "biochemical" assays, such as detecting the presence or absence of a particular peptide, e.g., by immunological means (ELISAs and Western blots) or by assays described herein to identify agents falling within the scope of the invention.

MHC class I proteins

[0330] MHC class I proteins are one of two primary classes of major histocompatibility complex (MHC) molecules (the other being MHC class II) and are found on nearly every nucleated cell of the body. Their function is to display fragments of proteins from within the cell to T cells; healthy cells will be ignored, while cells containing foreign proteins will be attacked by the immune system. Because MHC class I proteins present peptides derived from cytosolic proteins, the pathway of MHC class I presentation is often called the cytosolic or endogenous pathway. Class I MHC molecules bind peptides generated mainly from degradation of cytosolic proteins by the proteasome. The MHC Lpeptide complex is then inserted into the plasma membrane of the cell. The peptide is bound to the extracellular part of the class I MHC molecule. Thus, the function of the class I MHC is to display intracellular proteins to cytotoxic T cells (CTLs).

However, class I MHC can also present peptides generated from exogenous proteins, in a process known as cross-presentation.

[0331] MHC class I proteins consist of two polypeptide chains, a and p2-microglobulin (β2Μ). The two chains are linked noncovalently via interaction of b2m and the a3 domain. Only the a chain is polymorphic and encoded by a HLA gene, while the b2m subunit is not polymorphic and encoded by the β-2 microglobulin gene. The a3 domain is plasma membrane- spanning and interacts with the CD8 co-receptor of T-cells. The a3-CD8 interaction holds the MHC I molecule in place while the T cell receptor (TCR) on the surface of the cytotoxic T cell binds its al-a2 heterodimer ligand, and checks the coupled peptide for antigenicity. The al and a2 domains fold to make up a groove for peptides to bind. MHC class I proteins bind peptides that are 8-10 amino acid in length.

[0332] The human leukocyte antigen (HLA) genes are the human versions of the MHC genes. The three major MHC class I proteins in humans are HLA-A, HLA-B, and HLA-C, while the 3 minor ones are HLA-E, HLA-F, and HLA-G. HLA-A is ranked among the genes in humans with the fastest-evolving coding sequence. As of December 2013, there were 2432 known HLA-A alleles coding for 1740 active proteins and 117 null proteins. The HLA-A gene is located on the short arm of chromosome 6 and encodes the larger, a-chain, constituent of HLA-A. Variation of HLA-A a-chain is key to HLA function. This variation promotes genetic diversity in the population. Since each HLA has a different affinity for peptides of certain structures, greater variety of HLAs means greater variety of antigens to be 'presented' on the cell surface, enhancing the likelihood that a subset of the population will be resistant to any given foreign invader. This decreases the likelihood that a single pathogen has the capability to wipe out the entire human population. Each individual can express up to two types of HLA- A, one from each of their parents. Some individuals will inherit the same HLA-A from both parents, decreasing their individual HLA diversity; however, the majority of individuals will receive two different copies of HLA-A. This same pattern follows for all HLA groups. In other words, a person can only express either one or two of the 2432 known HLA-A alleles.

[0333] All alleles receive at least a four digit classification, e.g. , HLA-A*02: 12. The A signifies which HLA gene the allele belongs to. There are many HLA-A alleles, so that classification by serotype simplifies categorization. The next pair of digits indicates this assignment. For example, HLA-A*02:02, HLA-A*02:04, and HLA-A*02:324 are all members of the A2 serotype (designated by the *02 prefix). This group is the primary factor responsible for HLA compatibility. All numbers after this cannot be determined by serotyping and are designated through gene sequencing. The second set of digits indicates what HLA protein is produced. These are assigned in order of discovery and as of December 2013 there are 456 different HLA- A02 proteins known (assigned names HLA-A*02:01 to HLA-A*02:456). The shortest possible HLA name includes both of these details. Each extension beyond that signifies a nucleotide change that may or may not change the protein.

[0334] In some embodiments, the anti-RTMC antibody moiety specifically binds to a complex comprising an HIV- 1 RT peptide and an MHC class I protein, wherein the MHC class I protein is HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, or HLA-G. In some embodiments, the MHC class

I protein is HLA-A, HLA-B, or HLA-C. In some embodiments, the MHC class I protein is

HLA-A. In some embodiments, the MHC class I protein is HLA-B. In some embodiments, the

MHC class I protein is HLA-C. In some embodiments, the MHC class I protein is HLA-A01,

HLA-A02, HLA-A03, HLA-A09, HLA-A10, HLA- Al l, HLA-A19, HLA-A23, HLA-A24,

HLA-A25, HLA-A26, HLA-A28, HLA-A29, HLA-A30, HLA- A31, HLA-A32, HLA- A33,

HLA-A34, HLA- A36, HLA-A43, HLA-A66, HLA-A68, HLA-A69, HLA-A74, or HLA-A80. In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is any one of HLA-A*02:01-555, such as HLA-A*02:01, HLA-A*02:02, HLA-

A*02:03, HLA-A*02:04, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*02:08, HLA-

A*02:09, HLA-A*02: 10, HLA-A*02: 11, HLA-A*02: 12, HLA-A*02: 13, HLA-A*02: 14, HLA- A*02: 15, HLA-A*02: 16, HLA-A*02: 17, HLA-A*02: 18, HLA-A*02: 19, HLA-A*02:20, HLA- A*02:21, HLA-A*02:22, or HLA-A*02:24. In some embodiments, the MHC class I protein is HLA-A*02:01. HLA-A*02:01 is expressed in 39-46% of all Caucasians, and therefore represents a suitable choice of MHC class I protein for use in the present invention.

[0335] HIV-1 RT peptides suitable for use in generating anti-RTMC antibody moieties can be determined, for example, based on the presence of HLA-A*02:01 -binding motifs and cleavage sites for proteasomes and immune-proteasomes using computer prediction models known to those of skill in the art. For predicting MHC binding sites, such models include, but are not limited to, IEDB (Vita et al., The immune epitope database (IEDB) 3.0. Nucleic Acids Res. 2014 Oct 9. pii: gku938), ProPredl (described in more detail in Singh and Raghava, ProPred:

prediction ofHLA-DR binding sites. BIOINFORMATICS 17(12): 1236-1237, 2001), and SYFPEITHI (see Schuler et al. SYFPEITHI, Database for Searching and T-Cell Epitope Prediction, in Immunoinformatics Methods in Molecular Biology, vol 409(1): 75-93, 2007).

[0336] Once appropriate peptides have been identified, peptide synthesis may be done in accordance with protocols well known to those of skill in the art. Because of their relatively small size, the peptides of the invention may be directly synthesized in solution or on a solid support in accordance with conventional peptide synthesis techniques. Various automatic synthesizers are commercially available and can be used in accordance with known protocols. The synthesis of peptides in solution phase has become a well-established procedure for large- scale production of synthetic peptides and as such is a suitable alternative method for preparing the peptides of the invention (See for example, Solid Phase Peptide Synthesis by John Morrow Stewart and Martin et al. Application of Almez-mediated Amidation Reactions to Solution Phase Peptide Synthesis, Tetrahedron Letters Vol. 39, pages 1517-1520, 1998).

[0337] The binding activity of candidate HIV-1 RT peptides can be tested using the antigen- processing-deficient T2 cell line, which increases expression of HLA-A when stabilized by a peptide in the antigen-presenting groove. T2 cells are pulsed with the candidate peptide for a time sufficient to stabilize HLA-A expression on the cell surface, which can be measured using any methods known in the art, such as by immuno staining with a fluorescently labeled monoclonal antibody specific for HLA-A (for example, BB7.2) followed by fluorescence- activated cell-sorting (FACS) analysis. Preparation of anti-RTMC antibodies and anti-RTMC antibody moieties

[0338] In some embodiments, the anti-RTMC antibody or anti-RTMC antibody moiety is a monoclonal antibody. Monoclonal antibodies can be prepared, e.g., using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975) and Sergeeva et ah, Blood, 117(16):4262-4272, using the phage display methods described herein and in the Examples below, or using recombinant DNA methods {see, e.g., US Patent No. 4,816,567).

[0339] In a hybridoma method, a hamster, mouse, or other appropriate host animal is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes can be immunized in vitro. The immunizing agent can include a polypeptide or a fusion protein of the protein of interest, or a complex comprising at least two molecules, such as a complex comprising an HIV-1 RT peptide and an MHC class I protein. Generally, peripheral blood lymphocytes ("PBLs") are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell. See, e.g., Goding, Monoclonal Antibodies: Principles and Practice (New York: Academic Press, 1986), pp. 59-103. Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine, and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells can be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine ("HAT medium"), which prevents the growth of HGPRT-deficient cells.

[0340] In some embodiments, the immortalized cell lines fuse efficiently, support stable high- level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. In some embodiments, the immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, California and the American Type Culture Collection, Manassas, Virginia. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies. Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al. Monoclonal Antibody Production Techniques and Applications (Marcel Dekker, Inc.: New York, 1987) pp. 51-63. [0341] The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the polypeptide. The binding specificity of monoclonal antibodies produced by the hybridoma cells can be determined by

immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem., 107:220 (1980).

[0342] After the desired hybridoma cells are identified, the clones can be sub cloned by limiting dilution procedures and grown by standard methods. Goding, supra. Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.

[0343] The monoclonal antibodies secreted by the sub clones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0344] The anti-RTMC antibodies or antibody moieties may also be identified by screening combinatorial libraries for antibodies with the desired activity or activities. For example, a variety of methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing the desired binding characteristics. Such methods are reviewed, e.g., in Hoogenboom et al., Methods in Molecular Biology 178: 1-37 (O'Brien et al., ed., Human Press, Totowa, N.J., 2001) and further described, e.g., in McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581- 597 (1992); Marks and Bradbury, Methods in Molecular Biology 248: 161-175 (Lo, ed., Human Press, Totowa, N.J., 2003); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132(2004).

[0345] In certain phage display methods, repertoires of V_H and V_L genes are separately cloned by polymerase chain reaction (PCR) and recombined randomly in phage libraries, which can then be screened for antigen-binding phage as described in Winter et αί, Αηη. Rev. Immunol.,

12: 433-455 (1994). Phage typically display antibody fragments, either as single-chain Fv (scFv) fragments or as Fab fragments. Libraries from immunized sources provide high-affinity antibodies to the immunogen without the requirement of constructing hybridomas. Alternatively, the naive repertoire can be cloned {e.g., from human) to provide a single source of antibodies to a wide range of non-self and also self antigens without any immunization as described by Griffiths et al, EMBO J, 12: 725-734 (1993). Finally, naive libraries can also be made synthetically by cloning unrearranged V-gene segments from stem cells, and using PCR primers containing random sequence to encode the highly variable CDR3 regions and to accomplish rearrangement in vitro, as described by Hoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example: U.S. Pat. No. 5,750,373, and US Patent Publication Nos. 2005/0079574, 2005/0119455, 2005/0266000, 2007/0117126, 2007/0160598, 2007/0237764, 2007/0292936, and

2009/0002360.

[0346] The antibodies or antigen-binding fragments thereof can be prepared using phage display to screen libraries for antibodies specific to a complex comprising an HIV-1 RT peptide and an MHC class I protein. The library can be a human scFv phage display library having a diversity of at least one x 10⁹ (such as at least about any of 1 x 10⁹, 2.5 x 10⁹, 5 x 10⁹, 7.5 x 10⁹, 1 x 10¹⁰, 2.5 x 10¹⁰, 5 x 10¹⁰, 7.5 x 10¹⁰, or 1 x 10¹¹) unique human antibody fragments. In some embodiments, the library is a naive human library constructed from DNA extracted from human PMBCs and spleens from healthy donors, encompassing all human heavy and light chain subfamilies. In some embodiments, the library is a naive human library constructed from DNA extracted from PBMCs isolated from patients with various diseases, such as patients with autoimmune diseases, cancer patients, and patients with infectious diseases, such as HIV. In some embodiments, the library is a semi- synthetic human library, wherein heavy chain CDR3 is completely randomized, with all amino acids (with the exception of cysteine) equally likely to be present at any given position (see, e.g., Hoet, R.M. et al., Nat. Biotechnol. 23(3):344-348, 2005). In some embodiments, the heavy chain CDR3 of the semi- synthetic human library has a length from about 5 to about 24 (such as about any of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24) amino acids. In some embodiments, the library is a non-human phage display library.

[0347] Phage clones that bind to the RTMC with high affinity can be selected by iterative binding of phage to the RTMC, which is bound to a solid support (such as, for example, beads for solution panning or mammalian cells for cell panning), followed by removal of non-bound phage and by elution of specifically bound phage. In an example of solution panning, the RTMC can be biotinylated for immobilization to a solid support. The biotinylated RTMC is mixed with the phage library and a solid support, such as streptavidin-conjugated Dynabeads M-280, and then RTMC-phage-bead complexes are isolated. The bound phage clones are then eluted and used to infect an appropriate host cell, such as E. coli XLl-Blue, for expression and purification. In an example of cell panning, T2 cells (a TAP-deficient, HLA-A*02:01⁺ lymphoblast cell line) loaded with the HIV- 1 RT peptide of the RTMC are mixed with the phage library, after which the cells are collected and the bound clones are eluted and used to infect an appropriate host cell for expression and purification. The panning can be performed for multiple (such as about any of 2, 3, 4, 5, 6 or more) rounds with solution panning, cell panning, or a combination of both, to enrich for phage clones binding specifically to the RTMC. Enriched phage clones can be tested for specific binding to the RTMC by any methods known in the art, including for example ELISA and FACS.

[0348] Monoclonal antibodies can also be made by recombinant DNA methods, such as those described in U.S. Patent No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). Hybridoma cells as described above or RTMC-specific phage clones of the invention can serve as a source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. The DNA also can be modified, for example, by substituting the coding sequence for human heavy- and light-chain constant domains and/or framework regions in place of the homologous non-human sequences (U.S. Patent No. 4,816,567; Morrison et ah, supra) or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a nonimmunoglobulin polypeptide. Such a non- immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-combining site of an antibody of the invention to create a chimeric bivalent antibody.

[0349] The antibodies can be monovalent antibodies. Methods for preparing monovalent antibodies are known in the art. For example, one method involves recombinant expression of immunoglobulin light chain and modified heavy chain. The heavy chain is truncated generally at any point in the Fc region so as to prevent heavy-chain crosslinking. Alternatively, the relevant cysteine residues are substituted with another amino acid residue or are deleted so as to prevent crosslinking. [0350] In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce fragments thereof, particularly Fab fragments, can be accomplished using any method known in the art.

[0351] Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant-domain sequences. The fusion preferably is with an immunoglobulin heavy-chain constant domain, comprising at least part of the hinge, CH2, and CH3 regions. In some embodiments, the first heavy-chain constant region (C_HI) containing the site necessary for light-chain binding is present in at least one of the fusions. DNAs encoding the immunoglobulin heavy-chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co-transfected into a suitable host organism. For further details of generating bispecific antibodies, see, for example, Suresh et al., Methods in Enzymology, 121: 210 (1986).

Human and Humanized Antibodies

[0352] The anti-RTMC antibodies or antibody moieties can be humanized antibodies or human antibodies. Humanized forms of non-human (e.g., murine) antibodies are chimeric

immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F(ab')₂, scFv, or other antigen-binding subsequences of antibodies) that typically contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibody) in which residues from a CDR of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, or rabbit having the desired specificity, affinity, and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies can also comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody can comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin, and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. In some embodiments, the humanized antibody will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. See, e.g., Jones et al., Nature, 321: 522-525 (1986); Riechmann et al, Nature, 332: 323-329 (1988); Presta, Curr. Op. Struct. Biol, 2:593-596 (1992). [0353] Generally, a humanized antibody has one or more amino acid residues introduced into it from a source that is non-human. These non-human amino acid residues are often referred to as "import" residues, which are typically taken from an "import" variable domain. According to some embodiments, humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321: 522-525 (1986); Riechmann et al, Nature, 332: 323- 327 (1988); Verhoeyen et al, Science, 239: 1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such "humanized" antibodies are antibodies (U.S. Patent No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.

[0354] As an alternative to humanization, human antibodies can be generated. For example, it is now possible to produce transgenic animals (e.g., mice) that are capable, upon immunization, of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production. For example, it has been described that the homozygous deletion of the antibody heavy-chain joining region (JH) gene in chimeric and germ- line mutant mice results in complete inhibition of endogenous antibody production. Transfer of the human germ- line immunoglobulin gene array into such germ- line mutant mice will result in the production of human antibodies upon antigen challenge. See, e.g., Jakobovits et al, PNAS USA, 90:2551 (1993); Jakobovits et ah, Nature, 362:255-258 (1993); Bruggemann et ah, Year in Immunol., 7:33 (1993); U.S. Patent Nos. 5,545,806, 5,569,825, 5,591,669; 5,545,807; and WO 97/17852. Alternatively, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed that closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016, and Marks et al, Bio/Technology, 10: 779-783 (1992); Lonberg et al, Nature, 368: 856-859 (1994); Morrison, Nature, 368: 812-813 (1994); Fishwild et al, Nature Biotechnology, 14: 845-851 (1996); Neuberger, Nature Biotechnology, 14: 826 (1996); Lonberg and Huszar, Intern. Rev. Immunol, 13: 65-93 (1995).

[0355] Human antibodies may also be generated by in vitro activated B cells (see U.S. Patents

5,567,610 and 5,229,275) or by using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol, 227:381 (1991); Marks et al, J. Mol. Biol., 222:581 (1991). The techniques of Cole et al. and Boerner et al. are also available for the preparation of human monoclonal antibodies. Cole et al, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985) and Boerner et al, J. Immunol, 147(1): 86-95 (1991).

Multi-specific Antibodies

[0356] In some embodiments, the anti-RTMC construct is a multi- specific antibody. Suitable methods for making multi- specific {e.g., bispecific) antibodies are well known in the art. For example, the production of bispecific antibodies can based on the co-expression of two immunoglobulin heavy-chain/light-chain pairs, where the two pairs each have different specificities, and upon association result in a heterodimeric antibody {see, e.g., Milstein and Cuello, Nature, 305: 537-539 (1983); WO 93/08829, and Traunecker et al, EMBO J. 10: 3655 (1991)). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of ten different antibody molecules, of which only one has the correct bispecific structure. The purification of the correct molecule is usually accomplished by affinity chromatography steps. Similar procedures are disclosed in WO 93/08829 and in Traunecker et al, EMBO, 10: 3655-3659 (1991). Alternatively, the combining of heavy and light chains can be directed by taking advantage of species-restricted pairing {see, e.g., Lindhofer et al, J. Immunol, 155:219-225 (1995)) and the pairing of heavy chains can be directed by use of "knob-into hole" engineering of CH3 domains {see, e.g., U.S. Pat. No.

5,731,168; Ridgway et al, Protein Eng., 9(7):617-621 (1996)). Multi- specific antibodies may also be made by engineering electrostatic steering effects for making antibody Fc-heterodimeric molecules {see, e.g., WO 2009/089004A1). In yet another method, stable bispecific antibodies can be generated by controlled Fab-arm exchange, where two parental antibodies having distinct antigen specificity and matched point mutations in the CH3 domains are mixed in reducing condition to allow for separation, reassembly, and reoxidation to form highly pure bispecific antibodies. Labrigin et al, Proc. Natl. Acad. Scl, 110(13):5145-5150 (2013). Such antibodies, comprising a mixture of heavy-chain/light-chain pairs, are also referred to herein as

"heteromultimeric antibodies".

[0357] Antibodies or antigen-binding fragments thereof having different specificities can also be chemically cross-linked to generate multi- specific heteroconjugate antibodies. For example, two F(ab')2 molecules, each having specificity for a different antigen, can be chemically linked. Pullarkat et al, Trends Biotechnol, 48:9-21 (1999). Such antibodies have, for example, been proposed to target immune-system cells to unwanted cells (U.S. Patent No. 4,676,980), and for treatment of HIV infection. WO 91/00360; WO 92/200373; EP 03089. It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide-exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in U.S. Patent No. 4,676,980.

[0358] In some embodiments, multi- specific antibodies can be prepared using recombinant DNA techniques. For example, a bispecific antibody can be engineered by fusing two scFvs, such as by fusing them through a peptide linker, resulting in a tandem scFv. One example of a tandem scFv is a bispecific T cell engager. Bispecific T cell engagers are made by linking an anti-CD3 scFv to an scFv specific for a surface antigen of a target cell, such as a tumor- associated antigen (TAA), resulting in the redirection of T cells to the target cells. Mack et al., Proc. Natl. Acad. Sci., 92:7021-7025 (1995); Brischwein et al., Mol. Immunol, 43(8): 1129- 1143 (2006). By shortening the length of a peptide linker between two variable domains, they can be prevented from self-assembling and forced to pair with domains on a second polypeptide, resulting in a compact bispecific antibody called a diabody (Db). Holliger et al, Proc. Natl. Acad. Sci., 90:6444-6448 (1993). The two polypeptides of a Db each comprise a V_H connected to a V_L by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the V_H and V_L domains of one polypeptide are forced to pair with the complementary V_L and V_H domains of another polypeptide, thereby forming two antigen- binding sites. In a modification of this format, the two polypeptides are linked by another peptide linker, resulting in a single chain diabody (scDb). In yet another modification of the Db format, dual-affinity retargeting (DART) bispecific antibodies can be generated by introducing a disulfide linkage between cysteine residues at the C-terminus of each polypeptide, optionally including domains prior to the C-terminal cysteine residues that drive assembly of the desired heterodimeric structure. Veri et al, Arthritis Rheum., 62(7): 1933- 1943 (2010). Dual- variable- domain immunoglobulins (DVD-Ig™), in which the target-binding variable domains of two monoclonal antibodies are combined via naturally occurring linkers to yield a tetravalent, bispecific antibody, are also known in the art. Gu and Ghayur, Methods Enzymol, 502:25-41 (2012). In yet another format, Dock and Lock (DNL), bispecific antibodies are prepared by taking advantage of the dimerization of a peptide (DDD2) derived from the regulatory subunit of human cAMP-dependent protein kinase (PKA) with a peptide (AD2) derived from the anchoring domains of human A kinase anchor proteins (AKAPs). Rossi et ah, Proc. Natl. Acad. Sci., 103:6841-6846 (2006).

[0359] Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. Kostelny et ah, J. Immunol., 148(5): 1547-1553 (1992). This method can also be utilized for the production of antibody homodimers.

Anti-RTMC variants

[0360] In some embodiments, amino acid sequence variants of the antibody moieties provided herein are contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibody moiety. Amino acid sequence variants of an antibody moiety may be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody moiety, or by peptide synthesis. Such modifications include, for example, deletions from, and/or insertions into and/or substitutions of residues within the amino acid sequences of the antibody moiety. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen-binding.

[0361] In some embodiments, antibody moiety variants having one or more amino acid substitutions are provided. Sites of interest for substitutional mutagenesis include the HVRs and FRs. Amino acid substitutions may be introduced into an antibody moiety of interest and the products screened for a desired activity, e.g., retained/improved antigen binding, decreased immunogenicity, or improved ADCC or CDC.

[0362] Conservative substitutions are shown in Table 6 below.

TABLE 6: CONSERVATIVE SUBSTITITIONS

Gly (G) Ala Ala

His (H) Asn; Gin; Lys; Arg Arg

lie (I) Leu; Val; Met; Ala; Phe; Norleucine Leu

Leu (L) Norleucine; He; Val; Met; Ala; Phe He

Lys (K) Arg; Gin; Asn Arg

Met (M) Leu; Phe; He Leu

Phe (F) Trp; Leu; Val; He; Ala; Tyr Tyr

Pro (P) Ala Ala

Ser (S) Thr Thr

Thr (T) Val; Ser Ser

Trp (W) Tyr; Phe Tyr

Tyr (Y) Trp; Phe; Thr; Ser Phe

Val (V) He; Leu; Met; Phe; Ala; Norleucine Leu

[0363] Amino acids may be grouped into different classes according to common side-chain properties:

a. hydrophobic: Norleucine, Met, Ala, Val, Leu, He;

b. neutral hydrophilic: Cys, Ser, Thr, Asn, Gin;

c. acidic: Asp, Glu;

d. basic: His, Lys, Arg;

e. residues that influence chain orientation: Gly, Pro;

f. aromatic: Trp, Tyr, Phe.

[0364] Non-conservative substitutions will entail exchanging a member of one of these classes for another class.

[0365] An exemplary substitutional variant is an affinity matured antibody moiety, which may be conveniently generated, e.g., using phage display-based affinity maturation techniques. Briefly, one or more CDR residues are mutated and the variant antibody moieties displayed on phage and screened for a particular biological activity (e.g. binding affinity). Alterations (e.g., substitutions) may be made in HVRs, e.g., to improve antibody moiety affinity. Such alterations may be made in HVR "hotspots," i.e., residues encoded by codons that undergo mutation at high frequency during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol. 207: 179- 196 (2008)), and/or specificity determining residues (SDRs), with the resulting variant VH or VL being tested for binding affinity. Affinity maturation by constructing and reselecting from secondary libraries has been described, e.g., in Hoogenboom et al. in Methods in

Molecular Biology 178: 1-37 (O'Brien et ah, ed., Human Press, Totowa, NJ, (2001).)

[0366] In some embodiments of affinity maturation, diversity is introduced into the variable genes chosen for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then created. The library is then screened to identify any antibody moiety variants with the desired affinity.

Another method to introduce diversity involves HVR-directed approaches, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding may be specifically identified, e.g., using alanine scanning mutagenesis or modeling. CDR-H3 and CDR-L3 in particular are often targeted.

[0367] In some embodiments, substitutions, insertions, or deletions may occur within one or more HVRs so long as such alterations do not substantially reduce the ability of the antibody moiety to bind antigen. For example, conservative alterations (e.g. , conservative substitutions as provided herein) that do not substantially reduce binding affinity may be made in HVRs. Such alterations may be outside of HVR "hotspots" or SDRs. In some embodiments of the variant VH and VL sequences provided above, each HVR either is unaltered, or contains no more than one, two or three amino acid substitutions.

[0368] A useful method for identification of residues or regions of an antibody moiety that may be targeted for mutagenesis is called "alanine scanning mutagenesis" as described by

Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, a residue or group of target residues (e.g. , charged residues such as arg, asp, his, lys, and glu) are identified and replaced by a neutral or negatively charged amino acid (e.g. , alanine or polyalanine) to determine whether the interaction of the antibody moiety with antigen is affected. Further substitutions may be introduced at the amino acid locations demonstrating functional sensitivity to the initial substitutions. Alternatively, or additionally, a crystal structure of an antigen- antibody moiety complex can be determined to identify contact points between the antibody moiety and antigen. Such contact residues and neighboring residues may be targeted or eliminated as candidates for substitution. Variants may be screened to determine whether they contain the desired properties.

[0369] Amino acid sequence insertions include amino- and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an antibody moiety with an N-terminal methionyl residue. Other insertional variants of the antibody moiety include the fusion to the N- or C-terminus of the antibody moiety to an enzyme (e.g. for ADEPT) or a polypeptide which increases the serum half-life of the antibody moiety.

Fc Region Variants

[0370] In some embodiments, one or more amino acid modifications may be introduced into the Fc region of a full-length anti-RTMC antibody provided herein, thereby generating an Fc region variant. In some embodiments, the Fc region variant has enhanced antibody dependent cellular cytotoxicity (ADCC) effector function, often related to binding to Fc receptors (FcRs). In some embodiments, the Fc region variant has decreased ADCC effector function. There are many examples of changes or mutations to Fc sequences that can alter effector function. For example, WO 00/42072 and Shields et al. J Biol. Chem. 9(2): 6591-6604 (2001) describe antibody variants with improved or diminished binding to FcRs. The contents of those publications are specifically incorporated herein by reference.

[0371] Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) is a mechanism of action of therapeutic antibodies against tumor cells. ADCC is a cell-mediated immune defense whereby an effector cell of the immune system actively lyses a target cell (e.g., an HIV- 1 -infected cell), whose membrane- surface antigens have been bound by specific antibodies (e.g., an anti-RTMC antibody). The typical ADCC involves activation of NK cells by antibodies. An NK cell expresses CD16 which is an Fc receptor. This receptor recognizes, and binds to, the Fc portion of an antibody bound to the surface of a target cell. The most common Fc receptor on the surface of an NK cell is called CD16 or FcyRIII. Binding of the Fc receptor to the Fc region of an antibody results in NK cell activation, release of cytolytic granules and consequent target cell apoptosis. The contribution of ADCC to tumor cell killing can be measured with a specific test that uses NK-92 cells that have been transfected with a high-affinity FcR. Results are compared to wild-type NK-92 cells that do not express the FcR.

[0372] In some embodiments, the invention contemplates an anti-RTMC construct variant comprising an FC region that possesses some but not all effector functions, which makes it a desirable candidate for applications in which the half-life of the anti-RTMC construct in vivo is important yet certain effector functions (such as CDC and ADCC) are unnecessary or deleterious. In vitro and/or in vivo cytotoxicity assays can be conducted to confirm the reduction/depletion of CDC and/or ADCC activities. For example, Fc receptor (FcR) binding assays can be conducted to ensure that the antibody lacks FcyR binding (hence likely lacking ADCC activity), but retains FcRn binding ability. The primary cells for mediating ADCC, NK cells, express FcyRIII only, whereas monocytes express FcyRI, FcyRII and FcyRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest is described in U.S. Pat. No. 5,500,362 (see, e.g.

Hellstrom, I. et al. Proc. Nat'lAcad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I et al., Proc. Nat'l Acad. Sci. USA 82: 1499-1502 (1985); U.S. Pat. No. 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166: 1351-1361 (1987)). Alternatively, non-radioactive assay methods may be employed (see, for example, ACTI™ non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, Calif.; and CytoTox 96™ non-radioactive cytotoxicity assay (Promega, Madison, Wis.). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al. Proc. Nat'lAcad. Sci. USA 95:652-656 (1998). Clq binding assays may also be carried out to confirm that the antibody is unable to bind Clq and hence lacks CDC activity. See, e.g., Clq and C3c binding ELISA in WO 2006/029879 and WO 2005/100402. To assess complement activation, a CDC assay may be performed (see, for example, Gazzano- Santoro et al, J. Immunol. Methods 202: 163 (1996); Cragg, M. S. et al, Blood 101: 1045-1052 (2003); and Cragg, M. S. and M. J. Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance/half life determinations can also be performed using methods known in the art (see, e.g., Petkova, S. B. et al, Int'l. Immunol. 18(12): 1759-1769 (2006)).

[0373] Antibodies with reduced effector function include those with substitution of one or more of Fc region residues 238, 265, 269, 270, 297, 327 and 329 (U.S. Pat. No. 6,737,056). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called "DANA" Fc mutant with substitution of residues 265 and 297 to alanine (U.S. Pat. No. 7,332,581).

[0374] Certain antibody variants with improved or diminished binding to FcRs are described. (See, e.g., U.S. Pat. No. 6,737,056; WO 2004/056312, and Shields et al, J. Biol. Chem. 9(2): 6591-6604 (2001).)

[0375] In some embodiments, there is provided an anti-RTMC construct {e.g., a full-length anti-

RTMC antibody) variant comprising a variant Fc region comprising one or more amino acid substitutions which improve ADCC. In some embodiments, the variant Fc region comprises one or more amino acid substitutions which improve ADCC, wherein the substitutions are at positions 298, 333, and/or 334 of the variant Fc region (EU numbering of residues). In some embodiments, the anti-RTMC construct (e.g., full-length anti-RTMC antibody) variant comprises the following amino acid substitution in its variant Fc region: S298A, E333A, and K334A.

[0376] In some embodiments, alterations are made in the Fc region that result in altered (i.e., either improved or diminished) Clq binding and/or Complement Dependent Cytotoxicity (CDC), e.g., as described in U.S. Pat. No. 6,194,551, WO 99/51642, and Idusogie et al, J.

Immunol. 164: 4178-4184 (2000).

[0377] In some embodiments, there is provided an anti-RTMC construct (e.g., a full-length anti- RTMC antibody) variant comprising a variant Fc region comprising one or more amino acid substitutions which increase half-life and/or improve binding to the neonatal Fc receptor (FcRn). Antibodies with increased half-lives and improved binding to FcRn are described in

US2005/0014934A1 (Hinton et al). Those antibodies comprise an Fc region with one or more substitutions therein which improve binding of the Fc region to FcRn. Such Fc variants include those with substitutions at one or more of Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434, e.g., substitution of Fc region residue 434 (U.S. Pat. No. 7,371,826).

[0378] See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO 94/29351 concerning other examples of Fc region variants.

[0379] Anti-RTMC constructs (such as full-length anti-RTMC antibodies) comprising any of the Fc variants described herein, or combinations thereof, are contemplated.

Glycosylation Variants

[0380] In some embodiments, an anti-RTMC construct provided herein is altered to increase or decrease the extent to which the anti-RTMC construct is glycosylated. Addition or deletion of glycosylation sites to an anti-RTMC construct may be conveniently accomplished by altering the amino acid sequence of the anti-RTMC construct or polypeptide portion thereof such that one or more glycosylation sites is created or removed.

[0381] Where the anti-RTMC construct comprises an Fc region, the carbohydrate attached thereto may be altered. Native antibodies produced by mammalian cells typically comprise a branched, biantennary oligosaccharide that is generally attached by an N-linkage to Asn297 of the CH2 domain of the Fc region. See, e.g., Wright et al, TIBTECH 15:26-32 (1997). The oligosaccharide may include various carbohydrates, e.g., mannose, N-acetyl glucosamine (GlcNAc), galactose, and sialic acid, as well as a fucose attached to a GlcNAc in the "stem" of the biantennary oligosaccharide structure. In some embodiments, modifications of the oligosaccharide in an anti-RTMC construct of the invention may be made in order to create anti- RTMC construct variants with certain improved properties.

[0382] In some embodiments, anti-RTMC construct (such as full-length anti-RTMC antibody) variants are provided comprising an Fc region wherein a carbohydrate structure attached to the

Fc region has reduced fucose or lacks fucose, which may improve ADCC function. Specifically, anti-RTMC constructs are contemplated herein that have reduced fusose relative to the amount of fucose on the same anti-RTMC construct produced in a wild-type CHO cell. That is, they are characterized by having a lower amount of fucose than they would otherwise have if produced by native CHO cells (e.g., a CHO cell that produce a native glycosylation pattern, such as, a

CHO cell containing a native FUT8 gene). In some embodiments, the anti-RTMC construct is one wherein less than about 50%, 40%, 30%, 20%, 10%, or 5% of the N-linked glycans thereon comprise fucose. For example, the amount of fucose in such an anti-RTMC construct may be from 1% to 80%, from 1% to 65%, from 5% to 65% or from 20% to 40%. In some

embodiments, the anti-RTMC construct is one wherein none of the N-linked glycans thereon comprise fucose, i.e., wherein the anti-RTMC construct is completely without fucose, or has no fucose or is afucosylated. The amount of fucose is determined by calculating the average amount of fucose within the sugar chain at Asn297, relative to the sum of all glyco structures attached to

Asn 297 (e. g. complex, hybrid and high mannose structures) as measured by MALDI-TOF mass spectrometry, as described in WO 2008/077546, for example. Asn297 refers to the asparagine residue located at about position 297 in the Fc region (Eu numbering of Fc region residues); however, Asn297 may also be located about +3 amino acids upstream or downstream of position

297, i.e., between positions 294 and 300, due to minor sequence variations in antibodies. Such fucosylation variants may have improved ADCC function. See, e.g., US Patent Publication Nos.

US 2003/0157108 (Presta, L.); US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to "defucosylated" or "fucose-deficient" antibody variants include: US

2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614; US 2002/0164328; US

2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865;

WO 2003/085119; WO 2003/084570; WO 2005/035586; WO 2005/035778; WO2005/053742;

WO2002/031140; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al.

Biotech. Bioeng. 87: 614 (2004). Examples of cell lines capable of producing defucosylated antibodies include Lecl3 CHO cells deficient in protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US Pat Appl No US 2003/0157108 Al, Presta, L; and WO 2004/056312 Al, Adams et al., especially at Example 11), and knockout cell lines, such asa-l,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003/085107).

[0383] Anti-RTMC construct (such as full-length anti-RTMC antibody) variants are further provided with bisected oligosaccharides, e.g., in which a biantennary oligosaccharide attached to the Fc region of the anti-RTMC construct is bisected by GlcNAc. Such anti-RTMC construct (such as full-length anti-RTMC antibody) variants may have reduced fucosylation and/or improved ADCC function. Examples of such antibody variants are described, e.g., in WO 2003/011878 (Jean-Mairet et al.); U.S. Pat. No. 6,602,684 (Umana et al.); US 2005/0123546 (Umana et al.), and Ferrara et al., Biotechnology and Bioengineering, 93(5): 851-861 (2006). Anti-RTMC construct (such as full-length anti-RTMC antibody) variants with at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such anti- RTMC construct variants may have improved CDC function. Such antibody variants are described, e.g., in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S.); and WO

1999/22764 (Raju, S.).

[0384] In some embodiments, the anti-RTMC construct (such as full-length anti-RTMC antibody) variants comprising an Fc region are capable of binding to an FcyRIII. In some embodiments, the anti-RTMC construct (such as full-length anti-RTMC antibody) variants comprising an Fc region have ADCC activity in the presence of human effector cells or have increased ADCC activity in the presence of human effector cells compared to the otherwise same anti-RTMC construct (such as full-length anti-RTMC antibody) comprising a human wild- type IgGlFc region.

Cysteine Engineered Variants

[0385] In some embodiments, it may be desirable to create cysteine engineered anti-RTMC constructs (such as full-length anti-RTMC antibodies) in which one or more amino acid residues are substituted with cysteine residues. In some embodiments, the substituted residues occur at accessible sites of the anti-RTMC construct. By substituting those residues with cysteine, reactive thiol groups are thereby positioned at accessible sites of the anti-RTMC construct and may be used to conjugate the anti-RTMC construct to other moieties, such as drug moieties or linker-drug moieties, to create an anti-RTMC immunoconjugate, as described further herein. Cysteine engineered anti-RTMC constructs (such as full-length anti-RTMC antibodies) may be generated as described, e.g., in U.S. Pat. No. 7,521,541.

Derivatives

[0386] In some embodiments, an anti-RTMC construct provided herein may be further modified to contain additional nonproteinaceous moieties that are known in the art and readily available. The moieties suitable for derivatization of the anti-RTMC construct include but are not limited to water soluble polymers. Non-limiting examples of water soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly- 1,3-dioxo lane, poly-l,3,6-trioxane, ethylene/maleic anhydride copolymer, polyamino acids (either

homopolymers or random copolymers), and dextran or poly(n- vinyl pyrrolidone)polyethylene glycol, propropylene glycol homopolymers, prolypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The polymer may be of any molecular weight, and may be branched or unbranched. The number of polymers attached to the anti-RTMC construct may vary, and if more than one polymer are attached, they can be the same or different molecules. In general, the number and/or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular properties or functions of the anti-RTMC construct to be improved, whether the anti-RTMC construct derivative will be used in a therapy under defined conditions, etc.

[0387] In some embodiments, conjugates of an anti-RTMC construct and nonproteinaceous moiety that may be selectively heated by exposure to radiation are provided. In some

embodiments, the nonproteinaceous moiety is a carbon nanotube (Kam et ah, Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation may be of any wavelength, and includes, but is not limited to, wavelengths that do not harm ordinary cells, but which heat the

nonproteinaceous moiety to a temperature at which cells proximal to the anti-RTMC construct- nonproteinaceous moiety are killed.

Chimeric Receptor Effector Cell Preparation

[0388] The present invention in one aspect provides effector cells (such as lymphocytes, for example T cells) expressing an anti-RTMC chimeric receptor, such as an anti-RTMC CAR or anti-RTMC abTCR. Exemplary methods of preparing effector cells (such as T cells) expressing the anti-RTMC chimeric receptor (anti-RTMC chimeric receptor effector cells, such as anti- RTMC CAR T cells or anti-RTMC abTCR T cells) are provided herein.

[0389] In some embodiments, an anti-RTMC chimeric receptor (such as anti-RTMC CAR or anti-RTMC abTCR) effector cell (such as T cell) can be generated by introducing a vector (including for example a lentiviral vector) comprising a sequence encoding an anti-RTMC CAR (for example a CAR comprising an anti-RTMC antibody moiety and CD28 or 4- IBB and CD3ζ intracellular signaling sequences) or an anti-RTMC abTCR into the effector cell (such as T cell). In some embodiments, the anti-RTMC chimeric receptor effector cells (such as T cells) of the invention are able to replicate in vivo, resulting in long-term persistence that can lead to sustained control of an HIV-1 infection.

[0390] In some embodiments, the invention relates to administering a genetically modified T cell expressing an anti-RTMC CAR or anti-RTMC abTCR for the treatment of a patient having an HIV-1 infection or at risk of having an HIV-1 infection using lymphocyte infusion. In some embodiments, autologous lymphocyte infusion is used in the treatment. Autologous PBMCs are collected from a patient in need of treatment and T cells are activated and expanded using the methods described herein and known in the art and then infused back into the patient.

[0391] In some embodiments, the anti-RTMC CAR T cell expresses an anti-RTMC CAR comprising an anti-RTMC antibody moiety (also referred to herein as an "anti-RTMC CAR T cell"). In some embodiments, the anti-RTMC CAR T cell expresses an anti-RTMC CAR comprising an extracellular domain comprising the anti-RTMC antibody moiety and an intracellular domain comprising intracellular signaling sequences of CD3ζ and CD28 and/or 4-

1BB. In some embodiments, the anti-RTMC abTCR T cell expresses an anti-RTMC abTCR comprising an anti-RTMC antibody moiety (also referred to herein as an "anti-RTMC abTCR T cell"). In some embodiments, the anti-RTMC abTCR T cell expresses an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. The anti-RTMC chimeric receptor T cells (such as anti-RTMC CAR T cells or anti-RTMC abTCR T cells) of the invention can undergo robust in vivo T cell expansion and can establish RTMC- specific memory cells that persist at high levels for an extended amount of time in blood and bone marrow. In some embodiments, the anti-RTMC chimeric receptor T cells of the invention infused into a patient can eliminate RTMC-presenting cells, such as RTMC- presenting HIV-1 -infected cells, in vivo in patients having an HIV-1 infection. In some embodiments, the anti-RTMC chimeric receptor T cells of the invention infused into a patient can eliminate RTMC-presenting cells, such as RTMC-presenting HIV- 1 -infected cells, in vivo in patients having an HIV-1 infection that is refractory to at least one conventional treatment.

[0392] Prior to expansion and genetic modification of the T cells, a source of T cells is obtained from a subject. T cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In some embodiments of the present invention, any number of T cell lines available in the art may be used. In some embodiments of the present invention, T cells can be obtained from a unit of blood collected from a subject using any number of techniques known to the skilled artisan, such as Ficoll™ separation. In some embodiments, cells from the circulating blood of an individual are obtained by apheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets. In some embodiments, the cells collected by apheresis may be washed to remove the plasma fraction and to place the cells in an appropriate buffer or media for subsequent processing steps. In some embodiments, the cells are washed with phosphate buffered saline

(PBS). In some embodiments, the wash solution lacks calcium and may lack magnesium or may lack many if not all divalent cations. As those of ordinary skill in the art would readily appreciate a washing step may be accomplished by methods known to those in the art, such as by using a semi-automated "flow-through" centrifuge (for example, the Cobe 2991 cell processor, the Baxter CytoMate, or the Haemonetics Cell Saver 5) according to the

manufacturer's instructions. After washing, the cells may be resuspended in a variety of biocompatible buffers, such as Ca²⁺-free, Mg²⁺-free PBS, PlasmaLyte A, or other saline solutions with or without buffer. Alternatively, the undesirable components of the apheresis sample may be removed and the cells directly resuspended in culture media.

[0393] In some embodiments, T cells are isolated from peripheral blood lymphocytes by lysing the red blood cells and depleting the monocytes, for example, by centrifugation through a

PERCOLL™ gradient or by counterflow centrifugal elutriation. A specific subpopulation of T cells, such as CD3⁺, CD28⁺, CD4⁺, CD8⁺, CD45RA⁺, and CD45RO⁺ T cells, can be further isolated by positive or negative selection techniques. For example, in some embodiments, T cells are isolated by incubation with anti-CD3/anti-CD28 (i.e., 3x28)-conjugated beads, such as

DYNABEADS® M-450 CD3/CD28 T, for a time period sufficient for positive selection of the desired T cells. In some embodiments, the time period is about 30 minutes. In some embodiments, the time period ranges from 30 minutes to 36 hours or longer and all integer values there between. In some embodiments, the time period is at least one, 2, 3, 4, 5, or 6 hours. In some embodiments, the time period is 10 to 24 hours. In some embodiments, the incubation time period is 24 hours. For isolation of T cells from patients with leukemia, use of longer incubation times, such as 24 hours, can increase cell yield. Longer incubation times may be used to isolate T cells in any situation where there are few T cells as compared to other cell types, such as in isolating tumor infiltrating lymphocytes (TIL) from tumor tissue or from immune- compromised individuals. Further, use of longer incubation times can increase the efficiency of capture of CD8⁺ T cells. Thus, by simply shortening or lengthening the time T cells are allowed to bind to the CD3/CD28 beads and/or by increasing or decreasing the ratio of beads to T cells, subpopulations of T cells can be preferentially selected for or against at culture initiation or at other time points during the process. Additionally, by increasing or decreasing the ratio of anti- CD3 and/or anti-CD28 antibodies on the beads or other surface, subpopulations of T cells can be preferentially selected for or against at culture initiation or at other desired time points. The skilled artisan would recognize that multiple rounds of selection can also be used in the context of this invention. In some embodiments, it may be desirable to perform the selection procedure and use the "unselected" cells in the activation and expansion process. "Unselected" cells can also be subjected to further rounds of selection.

[0394] Enrichment of a T cell population by negative selection can be accomplished with a combination of antibodies directed to surface markers unique to the negatively selected cells. One method is cell sorting and/or selection via negative magnetic immunoadherence or flow cytometry that uses a cocktail of monoclonal antibodies directed to cell surface markers present on the cells negatively selected. For example, to enrich for CD4⁺ cells by negative selection, a monoclonal antibody cocktail typically includes antibodies to CD 14, CD20, CDl lb, CD 16, HLA-DR, and CD8. In some embodiments, it may be desirable to enrich for or positively select for regulatory T cells which typically express CD4⁺, CD25⁺, CD62Lhi, GITR⁺, and FoxP3⁺. Alternatively, in some embodiments, T regulatory cells are depleted by anti-CD25 conjugated beads or other similar methods of selection.

[0395] For isolation of a desired population of cells by positive or negative selection, the concentration of cells and surface (e.g. , particles such as beads) can be varied. In some embodiments, it may be desirable to significantly decrease the volume in which beads and cells are mixed together (i.e., increase the concentration of cells), to ensure maximum contact of cells and beads. For example, in some embodiments, a concentration of about 2 billion cells/ml is used. In some embodiments, a concentration of about 1 billion cells/ml is used. In some embodiments, greater than about 100 million cells/ml is used. In some embodiments, a concentration of cells of about any of 10, 15, 20, 25, 30, 35, 40, 45, or 50 million cells/ml is used. In some embodiments, a concentration of cells of about any of 75, 80, 85, 90, 95, or 100 million cells/ml is used. In some embodiments, a concentration of about 125 or about 150 million cells/ml is used. Using high concentrations can result in increased cell yield, cell activation, and cell expansion. Further, use of high cell concentrations allows more efficient capture of cells that may weakly express target antigens of interest, such as CD28-negative T cells, or from samples where there are many tumor cells present (i.e., leukemic blood, tumor tissue, etc.). Such populations of cells may have therapeutic value and would be desirable to obtain. For example, using high concentration of cells allows more efficient selection of CD8⁺ T cells that normally have weaker CD28 expression.

[0396] In some embodiments of the present invention, T cells are obtained from a patient directly following treatment. In this regard, it has been observed that following certain cancer treatments, in particular treatments with drugs that damage the immune system, shortly after treatment during the period when patients would normally be recovering from the treatment, the quality of T cells obtained may be optimal or improved for their ability to expand ex vivo.

Likewise, following ex vivo manipulation using the methods described herein, these cells may be in a preferred state for enhanced engraftment and in vivo expansion. Thus, it is contemplated within the context of the present invention to collect blood cells, including T cells, dendritic cells, or other cells of the hematopoietic lineage, during this recovery phase. Further, in some embodiments, mobilization (for example, mobilization with GM-CSF) and conditioning regimens can be used to create a condition in a subject wherein repopulation, recirculation, regeneration, and/or expansion of particular cell types is favored, especially during a defined window of time following therapy. Illustrative cell types include T cells, B cells, dendritic cells, and other cells of the immune system.

[0397] Whether prior to or after genetic modification of the T cells to express a desirable anti- RTMC chimeric receptor (such as anti-RTMC CAR or anti-RTMC abTCR), the T cells can be activated and expanded generally using methods as described, for example, in U.S. Pat. Nos. 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575;

7,067,318; 7,172,869; 7,232,566; 7,175,843; 5,883,223; 6,905,874; 6,797,514; 6,867,041; and U.S. Patent Application Publication No. 20060121005. [0398] Generally, the T cells of the invention are expanded by contact with a surface having attached thereto an agent that stimulates a CD3/TCR complex associated signal and a ligand that stimulates a co-stimulatory molecule on the surface of the T cells. In particular, T cell populations may be stimulated, such as by contact with an anti-CD3 antibody, or antigen- binding fragment thereof, or an anti-CD2 antibody immobilized on a surface, or by contact with a protein kinase C activator (e.g., bryostatin) in conjunction with a calcium ionophore. For co- stimulation of an accessory molecule on the surface of the T cells, a ligand that binds the accessory molecule is used. For example, a population of T cells can be contacted with an anti- CD3 antibody and an anti-CD28 antibody, under conditions appropriate for stimulating proliferation of the T cells. To stimulate proliferation of either CD4⁺ T cells or CD8⁺ T cells, an anti-CD3 antibody and an anti-CD28 antibody. Examples of an anti-CD28 antibody include 9.3, B-T3, XR-CD28 (Diaclone, Besancon, France) can be used as can other methods commonly known in the art (Berg et al., Transplant Proc. 30(8):3975-3977, 1998; Haanen et ah, J. Exp. Med. 190(9): 13191328, 1999; Garland et al, J. Immunol. Meth. 227(l-2):53-63, 1999).

Immunoconjugate preparation

[0399] The anti-RTMC immunoconjugates may be prepared using any methods known in the art. See, e.g., WO 2009/067800, WO 2011/133886, and U.S. Patent Application Publication No. 2014322129, incorporated by reference herein in their entirety.

[0400] The anti-RTMC antibody moiety of an anti-RTMC immunoconjugate may be "attached to" the effector molecule by any means by which the anti-RTMC antibody moiety can be associated with, or linked to, the effector molecule. For example, the anti-RTMC antibody moiety of an anti-RTMC immunoconjugate may be attached to the effector molecule by chemical or recombinant means. Chemical means for preparing fusions or conjugates are known in the art and can be used to prepare the anti-RTMC immunoconjugate. The method used to conjugate the anti-RTMC antibody moiety and effector molecule must be capable of joining the binding protein with the effector molecule without interfering with the ability of the binding protein to bind to the antigen on the target cell.

[0401] The anti-RTMC antibody moiety of an anti-RTMC immunoconjugate may be linked indirectly to the effector molecule. For example, the anti-RTMC antibody moiety of an anti- RTMC immunoconjugate may be directly linked to a liposome containing the effector molecule of one of several types. The effector molecule(s) and/or the anti-RTMC antibody moiety may also be bound to a solid surface. [0402] In some embodiments, the anti-RTMC antibody moiety of an anti-RTMC immunoconjugate and the effector molecule are both proteins and can be conjugated using techniques well known in the art. There are several hundred crosslinkers available that can conjugate two proteins. (See for example "Chemistry of Protein Conjugation and Crosslinking". 1991 , Shans Wong, CRC Press, Ann Arbor). The crosslinker is generally chosen based on the reactive functional groups available or inserted on the anti-RTMC antibody moiety and/or effector molecule. In addition, if there are no reactive groups, a photoactivatible crosslinker can be used. In certain instances, it may be desirable to include a spacer between the anti-RTMC antibody moiety and the effector molecule. Crosslinking agents known to the art include the homobifunctional agents: glutaraldehyde, dimethyladipimidate and Bis(diazobenzidine) and the heterobifunctional agents: m Maleimidobenzoyl-N-Hydroxysuccinimide and Sulfo-m

Maleimidobenzoyl-N-Hydroxysuccinimide.

[0403] In some embodiments, the anti-RTMC antibody moiety of an anti-RTMC

immunoconjugate may be engineered with specific residues for chemical attachment of the effector molecule. Specific residues used for chemical attachment of molecule known to the art include lysine and cysteine. The crosslinker is chosen based on the reactive functional groups inserted on the anti-RTMC antibody moiety, and available on the effector molecule.

[0404] An anti-RTMC immunoconjugate may also be prepared using recombinant DNA techniques. In such a case a DNA sequence encoding the anti-RTMC antibody moiety is fused to a DNA sequence encoding the effector molecule, resulting in a chimeric DNA molecule. The chimeric DNA sequence is transfected into a host cell that expresses the fusion protein. The fusion protein can be recovered from the cell culture and purified using techniques known in the art.

[0405] Examples of attaching an effector molecule, which is a label, to the binding protein include the methods described in Hunter, et al, Nature 144:945 (1962); David, et al,

Biochemistry 13: 1014 (1974); Pain, et al, J. Immunol. Meth. 40:219 (1981); Nygren, J.

Histochem. and Cytochem. 30:407 (1982); Wensel and Meares, Radioimmunoimaging And Radioimmunotherapy, Elsevier, N.Y. (1983); and Colcher et al, "Use Of Monoclonal

Antibodies As Radiopharmaceuticals For The Localization Of Human Carcinoma Xenografts In Athymic Mice", Meth. Enzymol, 121:802-16 (1986).

[0406] The radio- or other labels may be incorporated in the immunoconjugate in known ways.

For example, the peptide may be biosynthesized or may be synthesized by chemical amino acid synthesis using suitable amino acid precursors involving, for example, fluorine- 19 in place of hydrogen. Labels such as ⁹⁹Tc or ¹²³I, ¹⁸⁶Re, ¹⁸⁸Re and ^{U 1}ln can be attached via a cysteine residue in the peptide. Yttrium-90 can be attached via a lysine residue. The IODOGEN method (Fraker et ah, Biochem. Biophys. Res. Commun. 80:49-57 (1978)) can be used to incorporate iodine-123. "Monoclonal Antibodies in Immuno scintigraphy" (Chatal, CRC Press 1989) describes other methods in detail.

[0407] Immunoconjugates of the antibody moiety and a cytotoxic agent may be made using a variety of bifunctional protein coupling agents such as N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP), succinimidyl-4-(N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCI), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)- ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1 ,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et ah, Science 238: 1098 (1987).

Carbon- 14-labeled l-isothiocyanatobenzyl-3-methyldiethylene tnaminepentaacetic acid (MX- DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See, e.g., WO94/11026. The linker may be a "cleavable linker" facilitating release of the cytotoxic drug in the cell. For example, an acid-labile linker, peptidase- sensitive linker, photolabile linker, dimethyl linker or disulfide-containing linker (Chari et ah, Cancer Research 52: 127-131 (1992); U.S. Patent No. 5,208,020) may be used.

[0408] The anti-RTMC immunoconjugates of the invention expressly contemplate, but are not limited to, ADC prepared with cross-linker reagents: BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo- KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, and sulfo-SMPB, and SVSB (succinimidyl-(4- vinylsulfone)benzoate) which are commercially available {e.g., from Pierce Biotechnology, Inc., Rockford, IL, U.S.A). See pages 467-498, 2003-2004 Applications Handbook and Catalog.

Pharmaceutical Compositions

[0409] Also provided herein are compositions (such as pharmaceutical compositions, also referred to herein as formulations) comprising an anti-RTMC construct. In some embodiments, the composition further comprises a cell (such as an effector cell, e.g., a T cell) associated with the anti-RTMC construct. In some embodiments, there is provided a pharmaceutical composition comprising an anti-RTMC construct and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition further comprises a cell (such as an effector cell, e.g., a T cell) associated with the anti-RTMC construct.

[0410] Suitable formulations of the anti-RTMC constructs are obtained by mixing an anti- RTMC construct having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as olyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter- ions such as sodium; metal complexes (e.g. Zn- protein complexes); and/or no n- ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). Exemplary formulations are described in W098/56418, expressly incorporated herein by reference. Lyophilized formulations adapted for subcutaneous

administration are described in WO97/04801. Such lyophilized formulations may be

reconstituted with a suitable diluent to a high protein concentration and the reconstituted formulation may be administered subcutaneously to the individual to be treated herein.

Lipofectins or liposomes can be used to deliver the anti-RTMC constructs of this invention into cells.

[0411] The formulation herein may also contain one or more active compounds in addition to the anti-RTMC construct as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. For example, it may be desirable to further provide an anti-neoplastic agent, a growth inhibitory agent, a cytotoxic agent, or a chemotherapeutic agent in addition to the anti-RTMC construct. Such molecules are suitably present in combination in amounts that are effective for the purpose intended. The effective amount of such other agents depends on the amount of anti-RTMC construct present in the formulation, the stage of HIV- 1 infection, and other factors discussed above. These are generally used in the same dosages and with administration routes as described herein or about from 1 to 99% of the heretofore employed dosages.

[0412] The anti-RTMC constructs may also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example,

hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Sustained- release preparations may be prepared.

[0413] Sustained-release preparations of the anti-RTMC constructs can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody (or fragment thereof), which matrices are in the form of shaped articles, e.g., films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate ), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT TM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydro gels release proteins for shorter time periods. When encapsulated antibodies remain in the body for a long time, they can denature or aggregate as a result of exposure to moisture at 37°C, resulting in a loss of biological activity and possible changes in

immunogenicity. Rational strategies can be devised for stabilization of anti-RTMC constructs depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thio-disulfide interchange, stabilization can be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.

[0414] In some embodiments, the anti-RTMC construct is formulated in a buffer comprising a citrate, NaCl, acetate, succinate, glycine, polysorbate 80 (Tween 80), or any combination of the foregoing. In some embodiments, the anti-RTMC construct is formulated in a buffer comprising about 100 mM to about 150 mM glycine. In some embodiments, the anti-RTMC construct is formulated in a buffer comprising about 50mM to about 100 mM NaCl. In some embodiments, the anti-RTMC construct is formulated in a buffer comprising about lOmM to about 50 mM acetate. In some embodiments, the anti-RTMC construct is formulated in a buffer comprising about lOmM to about 50 mM succinate. In some embodiments, the anti-RTMC construct is formulated in a buffer comprising about 0.005% to about 0.02% polysorbate 80. In some embodiments, the anti-RTMC construct is formulated in a buffer having a pH between about 5.1 and 5.6. In some embodiments, the anti-RTMC construct is formulated in a buffer comprising 10 mM citrate, 100 mM NaCl, lOOmM glycine, and 0.01% polysorbate 80, wherein the formulation is at pH 5.5.

[0415] The formulations to be used for in vivo administration must be sterile. This is readily accomplished by, e.g., filtration through sterile filtration membranes.

Methods for treatment using anti-RTMC constructs

[0416] The anti-RTMC constructs and/or compositions of the invention can be administered to individuals (e.g., mammals such as humans) to treat an HIV-1 infection. The present application thus in some embodiments provides a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition (such as a pharmaceutical composition) comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety, such as any one of the anti-RTMC constructs described herein. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-RTMC construct.

[0417] For example, in some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein. In some embodiments, the HIV-1 RT peptide comprises (such as consists of) the amino acid sequence of any one of SEQ ID NOs: 5-18. In some embodiments, the HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full- length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-RTMC construct. In some embodiments, the individual is human.

[0418] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-RTMC construct. In some embodiments, the individual is human.

[0419] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, an HC-

CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions. In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247- 249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250- 253. In some embodiments, the anti-RTMC construct is non- naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-RTMC construct. In some embodiments, the individual is human.

[0420] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-

124, or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; and an HC-CDR3 comprising the amino acid sequence of any one of

SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (for example about any of

1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-

189, or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions; an LC-CDR2 comprising the amino acid sequence of any one of SEQ

ID NOs: 190-207, or a variant thereof comprising up to about 3 (for example about any of 1, 2, or 3) amino acid substitutions; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, there is provided a method of treating an HIV- 1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti- RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (for example about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences. In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises: i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164- 189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti- RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-RTMC construct. In some embodiments, the individual is human.

[0421] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an anti-RTMC construct comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, wherein the anti-RTMC antibody moiety comprises a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full-length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-RTMC construct. In some embodiments, the individual is human.

[0422] For example, in some embodiments, the anti-RTMC construct contained in a

composition administered to an individual in a method of treating an HIV-1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165, 191, and 209, respectively, SEQ ID NOs: 77, 99,

respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and 214,

[0423] In some embodiments, the anti-RTMC construct contained in a composition administered to an individual in a method of treating an HIV- 1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-

CDR2, HC-CDR3, LC-CDR1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165,

191, and 209, respectively, SEQ ID NOs: 77, 99, 127, 164, 192, and 210, respectively, SEQ ID

NOs: 78, 100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81, 103, 131, 169, 191, and 214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105, 134, 164, 192, and 217, respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218,

respectively, SEQ ID NOs: 84, 107, 136, 172, 197, and 219, respectively, SEQ ID NOs: 85, 108, 137, 169, 191, and 218, respectively, SEQ ID NOs: 86, 109, 138, 173, 198, and 220,

[0424] In some embodiments, the anti-RTMC construct contained in a composition administered to an individual in a method of treating an HIV- 1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-

CDR2, HC-CDR3, LC-CDR 1, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98, 126, 165,

NOs: 78, 100, 128, 166, 193, and 211, respectively, SEQ ID NOs: 79, 101, 129, 167, 194, and 212, respectively, SEQ ID NOs: 80, 102, 130, 168, 192, and 213, respectively, SEQ ID NOs: 81,

103, 131, 169, 191, and 214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105,

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively.

[0425] In some embodiments, the anti-RTMC construct contained in a composition administered to an individual in a method of treating an HIV- 1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ

ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and

51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively,

SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ

ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively.

[0426] In some embodiments, the anti-RTMC construct contained in a composition administered to an individual in a method of treating an HIV- 1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity.

In some embodiments, the anti-RTMC construct contained in a composition administered to an individual in a method of treating an HIV-1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of

SEQ ID NOs: 27 and 55, respectively. In some embodiments, the anti-RTMC construct contained in a composition administered to an individual in a method of treating an HIV-1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC construct contained in a composition administered to an individual in a method of treating an HIV- 1 infection comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

[0427] In some embodiments of any of the methods for treating an HIV- 1 infection described above, the anti-RTMC construct is conjugated to a cell (such as an immune cell, e.g. , a T cell) prior to being administered to the individual. Thus, for example, there is provided a method of treating an HIV- 1 infection in an individual comprising a) conjugating any one of the anti- RTMC constructs described herein to a cell (such as an immune cell, e.g. , a T cell) to form an anti-RTMC construct/cell conjugate, and b) administering to the individual an effective amount of a composition comprising the anti-RTMC construct/cell conjugate. In some embodiments, the cell is derived from the individual. In some embodiments, the cell is not derived from the individual. In some embodiments, the anti-RTMC construct is conjugated to the cell by covalent linkage to a molecule on the surface of the cell. In some embodiments, the anti-RTMC construct is conjugated to the cell by non-covalent linkage to a molecule on the surface of the cell. In some embodiments, the anti-RTMC construct is conjugated to the cell by insertion of a portion of the anti-RTMC construct into the outer membrane of the cell. In some embodiments, the anti-RTMC construct is non-naturally occurring. In some embodiments, the anti-RTMC construct is a full- length antibody. In some embodiments, the anti-RTMC construct is a multi- specific (such as bispecific) molecule. In some embodiments, the anti-RTMC construct is a chimeric antigen receptor. In some embodiments, the anti-RTMC construct is an immunoconjugate. In some embodiments, the individual is human.

[0428] In some embodiments, the individual is a mammal (e.g., human, non- human primate, rat, mouse, cow, horse, pig, sheep, goat, dog, cat, etc.). In some embodiments, the individual is a human. In some embodiments, the individual is a clinical patient, a clinical trial volunteer, an experimental animal, etc. In some embodiments, the individual is younger than about 60 years old (including for example younger than about any of 50, 40, 30, 25, 20, 15, or 10 years old). In some embodiments, the individual is older than about 60 years old (including for example older than about any of 70, 80, 90, or 100 years old). In some embodiments, the individual is diagnosed with HIV- 1 infection.

[0429] The present application in some embodiments provides a method of delivering an anti- RTMC construct (such as any one of the anti-RTMC constructs described herein) to a cell presenting on its surface a complex comprising an HIV- 1 RT peptide and an MHC class I protein in an individual, the method comprising administering to the individual a composition comprising the anti-RTMC construct. In some embodiments, the anti-RTMC construct to be delivered is associated with a cell (such as an effector cell, e.g. , a T cell).

[0430] Many diagnostic methods for HIV- 1 infection are known in the art. Such methods include, but are not limited to, e.g., immunohistochemistry, PCR, and fluorescent in situ hybridization (FISH).

[0431] In some embodiments, the anti-RTMC constructs and/or compositions of the invention are administered in combination with a second, third, or fourth agent (including, e.g., an antiviral drug) to treat HIV- 1 infection. In some embodiments, the anti-RTMC construct is administered in combination with an agent that increases the expression of MHC class I proteins and/or enhances the surface presentation of HIV- 1 RT peptides by MHC class I proteins. In some embodiments, the agent includes, for example, IFN receptor agonists, Hsp90 inhibitors, enhancers of p53 expression, and chemotherapeutic agents. In some embodiments, the agent is an IFN receptor agonist including, for example, IFNy, IFNP, and IFNa. In some embodiments, the agent is an Hsp90 inhibitor including, for example, tanespimycin (17-AAG), alvespimycin (17-DMAG), retaspimycin (IPI-504), IPI-493, CNF2024/BIIB021, MPC-3100, Debio 0932 (CUDC-305), PU-H71, Ganetespib (STA-9090), NVP-AUY922 (VER-52269), HSP990, KW- 2478, AT13387, SNX-5422, DS-2248, and XL888. In some embodiments, the agent is an enhancer of p53 expression including, for example, 5-fluoro uracil and nutlin-3. In some embodiments, the agent is a chemotherapeutic agent including, for example, topotecan, etoposide, cisplatin, paclitaxel, and vinblastine.

[0432] In some embodiments, there is provided a method of treating an HIV- 1 infection in an individual, wherein the cells expressing HIV- 1 RT do not normally present, or present at relatively low levels, a complex comprising an HIV- 1 RT protein and an MHC class I protein on their surface, the method comprising administering to the individual a composition comprising an anti-RTMC construct in combination with an agent that increases the expression of MHC class I proteins and/or enhances the surface presentation of HIV- 1 RT peptides by MHC class I proteins. In some embodiments, the agent includes, for example, IFN receptor agonists, Hsp90 inhibitors, enhancers of p53 expression, and chemotherapeutic agents. In some embodiments, the agent is an IFN receptor agonist including, for example, IFNy, IFNP, and IFNa. In some embodiments, the agent is an Hsp90 inhibitor including, for example, tanespimycin (17-AAG), alvespimycin (17-DMAG), retaspimycin (IPI-504), IPI-493, CNF2024/BIIB021, MPC-3100,

Debio 0932 (CUDC-305), PU-H71, Ganetespib (STA-9090), NVP-AUY922 (VER-52269), HSP990, KW-2478, AT13387, SNX-5422, DS-2248, and XL888. In some embodiments, the agent is an enhancer of p53 expression including, for example, 5-fluorouracil and nutlin-3. In some embodiments, the agent is a chemotherapeutic agent including, for example, topotecan, etoposide, cisplatin, paclitaxel, and vinblastine.

[0433] Cancer treatments can be evaluated, for example, by tumor regression, tumor weight or size shrinkage, time to progression, duration of survival, progression free survival, overall response rate, duration of response, quality of life, protein expression and/or activity.

Approaches to determining efficacy of the therapy can be employed, including for example, measurement of response through radiological imaging.

[0434] In some embodiments, the efficacy of treatment is measured as the percentage tumor growth inhibition (% TGI), calculated using the equation 100-(T/C x 100), where T is the mean relative tumor volume of the treated tumor, and C is the mean relative tumor volume of a non- treated tumor. In some embodiments, the %TGI is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94% , about 95%, or more than 95%.

Dosing and Method of Administering the anti-RTMC construct Compositions

[0435] The dose of the anti-RTMC construct compositions administered to an individual (such as a human) may vary with the particular composition, the mode of administration, and the stage of the HIV-1 infection. In some embodiments, the amount of the composition is effective to result in an undetectable viral load. In some embodiments, the amount of the anti-RTMC construct composition is sufficient to result in a complete elimination of latent HIV reservoirs.

[0436] In some embodiments, the amount of the composition is sufficient to prolong overall survival of the individual. In some embodiments, the amount of the composition (for example when administered along) is sufficient to produce clinical benefit of more than about any of 50%, 60%, 70%, or 77% among a population of individuals treated with the anti-RTMC construct composition.

[0437] In some embodiments, the amount of the composition, alone or in combination with a second, third, and/or fourth agent, is an amount sufficient to decrease the viral load or decrease the number of HIV-1 -infected cells by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 100% compared to the corresponding viral load or number of HIV- 1- infected cells in the same subject prior to treatment or compared to the corresponding values in other subjects not receiving the treatment. Standard methods can be used to measure the magnitude of this effect, such as in vitro assays with purified enzyme, cell-based assays, animal models, or human testing.

[0438] In some embodiments, the amount of the anti-RTMC construct {e.g., full-length anti- RTMC antibody, multi- specific anti-RTMC molecule, anti-RTMC CAR, anti-RTMC abTCR, or anti-RTMC immunoconjugate) in the composition is below the level that induces a toxicological effect {i.e., an effect above a clinically acceptable level of toxicity) or is at a level where a potential side effect can be controlled or tolerated when the composition is administered to the individual.

[0439] In some embodiments, the amount of the composition is close to a maximum tolerated dose (MTD) of the composition following the same dosing regimen. In some embodiments, the amount of the composition is more than about any of 80%, 90%, 95%, or 98% of the MTD.

[0440] In some embodiments, the amount of an anti-RTMC construct {e.g., full-length anti- RTMC antibody, multi- specific anti-RTMC molecule, anti-RTMC CAR, anti-RTMC abTCR, or anti-RTMC immunoconjugate) in the composition is included in a range of about 0.001 μg to about 1000 μg.

[0441] In some embodiments of any of the above aspects, the effective amount of an anti-RTMC construct {e.g., full-length anti-RTMC antibody, multi- specific anti-RTMC molecule, anti- RTMC CAR, anti-RTMC abTCR, or anti-RTMC immunoconjugate) in the composition is in the range of about 0.1 g/kg to about 100 mg/kg of total body weight.

[0442] The anti-RTMC construct compositions can be administered to an individual (such as human) via various routes, including, for example, intravenous, intra- arterial, intraperitoneal, intrapulmonary, oral, inhalation, intravesicular, intramuscular, intra-tracheal, subcutaneous, intraocular, intrathecal, transmucosal, and transdermal. In some embodiments, sustained continuous release formulation of the composition may be used. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered intraportally. In some embodiments, the composition is administered

intraarterially. In some embodiments, the composition is administered intraperitoneally. In some embodiments, the composition is administered intrahepatically. In some embodiments, the composition is administered by hepatic arterial infusion.

Anti-RTMC Chimeric Receptor Effector Cell Therapy

[0443] The present application also provides methods of using an anti-RTMC chimeric receptor (such as an anti-RTMC CAR or anti-RTMC abTCR) to redirect the specificity of an effector cell (such as a primary T cell) to a complex comprising an HIV-1 RT peptide and an MHC class I protein. Thus, the present invention also provides a method of stimulating an effector cell- mediated response (such as a T cell-mediated immune response) to a target cell population or tissue comprising RTMC-presenting cells in a mammal, comprising the step of administering to the mammal an effector cell (such as a T cell) that expresses an anti-RTMC CAR or anti-RTMC abTCR.

[0444] Anti-RTMC chimeric receptor effector cells (such as anti-RTMC CAR T cells or anti- RTMC abTCR T cells) expressing the anti-RTMC chimeric receptor can be infused to a recipient in need thereof. The infused cell is able to kill RTMC-presenting cells in the recipient. In some embodiments, unlike antibody therapies, anti-RTMC chimeric receptor effector cells (such as T cells) are able to replicate in vivo resulting in long-term persistence that can lead to sustained tumor control.

[0445] In some embodiments, the anti-RTMC chimeric receptor effector cells are anti-RTMC CAR T cells or anti-RTMC abTCR T cells that can undergo robust in vivo T cell expansion and can persist for an extended amount of time. In some embodiments, the anti-RTMC CAR T cells or anti-RTMC abTCR T cells of the invention develop into specific memory T cells that can be reactivated to inhibit any additional tumor formation or growth.

[0446] The anti-RTMC chimeric receptor T cells (such as anti-RTMC CAR T cells or anti- RTMC abTCR T cells) of the invention may also serve as a type of vaccine for ex vivo immunization and/or in vivo therapy in a mammal. In some embodiments, the mammal is a human.

[0447] With respect to ex vivo immunization, of least one of the following occurs in vitro prior to administering the cell into a mammal: i) expansion of the cells, ii) introducing a nucleic acid encoding an anti-RTMC CAR or anti-RTMC abTCR to the cells, and/or iii) cryopreservation of the cells.

[0448] Ex vivo procedures are well known in the art and are discussed more fully below. Briefly, cells are isolated from a mammal (preferably a human) and genetically modified (i.e., transduced or transfected in vitro) with a vector expressing an anti-RTMC CAR or anti-RTMC abTCR disclosed herein. The anti-RTMC CAR cell or anti-RTMC abTCR cell can be administered to a mammalian recipient to provide a therapeutic benefit. The mammalian recipient may be a human and the anti-RTMC CAR or anti-RTMC abTCR cell can be autologous with respect to the recipient. Alternatively, the cells can be allogeneic, syngeneic or xenogeneic with respect to the recipient. [0449] The procedure for ex vivo expansion of hematopoietic stem and progenitor cells is described in U.S. Pat. No. 5,199,942, incorporated herein by reference, can be applied to the cells of the present invention. Other suitable methods are known in the art, therefore the present invention is not limited to any particular method of ex vivo expansion of the cells. Briefly, ex vivo culture and expansion of T cells comprises: (1) collecting CD34⁺ hematopoietic stem and progenitor cells from a mammal from peripheral blood harvest or bone marrow explants; and (2) expanding such cells ex vivo. In addition to the cellular growth factors described in U.S. Pat. No. 5,199,942, other factors such as flt3-L, IL-1, IL-3 and c-kit ligand, can be used for culturing and expansion of the cells.

[0450] In addition to using a cell-based vaccine in terms of ex vivo immunization, the present invention also provides compositions and methods for in vivo immunization to elicit an immune response directed against an antigen in a patient.

[0451] The anti-RTMC chimeric receptor effector cells (such as anti-RTMC CAR T cells or anti-RTMC abTCR T cells) of the present invention may be administered either alone, or as a pharmaceutical composition in combination with diluents and/or with other components such as IL-2 or other cytokines or cell populations. Briefly, pharmaceutical compositions of the present invention may comprise anti-RTMC chimeric receptor effector cells (such as T cells), in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. Such compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants;

chelating agents such as EDTA or glutathione; adjuvants {e.g., aluminum hydroxide); and preservatives. In some embodiments, anti-RTMC chimeric receptor effector cell (such as T cell) compositions are formulated for intravenous administration.

[0452] The precise amount of the anti-RTMC chimeric receptor effector cell (such as anti- RTMC CAR T cell or anti-RTMC abTCR T cell) compositions of the present invention to be administered can be determined by a physician with consideration of individual differences in age, weight, tumor size, extent of infection or metastasis, and condition of the patient (subject). In some embodiments, a pharmaceutical composition comprising the anti-RTMC chimeric receptor effector cells (such as T cells) is administered at a dosage of about 10⁴ to about 10⁹ cells/kg body weight, such any of about 10⁴ to about 10⁵, about 10⁵ to about 10⁶, about 10⁶ to about 10 7 , about 107 to about 108 , or about 108 to about 109 cells/kg body weight, including all integer values within those ranges. Anti-RTMC chimeric receptor effect cell (such as T cell) compositions may also be administered multiple times at these dosages. The cells can be administered by using infusion techniques that are commonly known in immunotherapy (see, e.g., Rosenberg et al., New Eng. J. of Med. 319: 1676, 1988). The optimal dosage and treatment regimen for a particular patient can readily be determined by one skilled in the art of medicine by monitoring the patient for signs of disease and adjusting the treatment accordingly.

[0453] In some embodiments, it may be desired to administer activated anti-RTMC chimeric receptor T cells (such as anti-RTMC CAR T cells or anti-RTMC abTCR T cells) to a subject and then subsequently redraw blood (or have an apheresis performed), activate T cells therefrom according to the present invention, and reinfuse the patient with these activated and expanded T cells. This process can be carried out multiple times every few weeks. In some embodiments, T cells can be activated from blood draws of from 10 cc to 400 cc. In some embodiments, T cells are activated from blood draws of 20 cc, 30 cc, 40 cc, 50 cc, 60 cc, 70 cc, 80 cc, 90 cc, or 100 cc.

[0454] The administration of the anti-RTMC chimeric receptor effector cells (such as anti- RTMC CAR T cells or anti-RTMC abTCR T cells) may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. The compositions described herein may be administered to a patient

subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous (i.v.) injection, or intraperitoneally. In some embodiments, the anti-RTMC chimeric receptor effector cell (such as T cell) compositions of the present invention are administered to a patient by intradermal or subcutaneous injection. In some embodiments, the anti-RTMC chimeric receptor effector cell (such as T cell) compositions of the present invention are administered by i.v. injection. The compositions of anti-RTMC chimeric receptor effector cells (such as T cells) may be injected directly into a tumor, lymph node, or site of infection.

[0455] Thus, for example, in some embodiments, there is provided a method of treating an HIV- 1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ

ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the individual is human.

[0456] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0457] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1,

3) amino acid substitutions, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0458] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0459] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2,

3, 4, or 5) amino acid substitutions; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human. [0460] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0461] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human. [0462] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ

intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0463] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC CAR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47- 74; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0464] For example, in some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-

CDR1, HC-CDR2, HC-CDR3, LC-CDRl, LC-CDR2, and LC-CDR3 comprising the amino acid sequences of SEQ ID NOs: 75, 97, 125, 164, 190, and 208, respectively, SEQ ID NOs: 76, 98,

[0465] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

[0466] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

137, 169, 191, and 218, respectively, SEQ ID NOs: 86, 109, 138, 173, 198, and 220, respectively, SEQ ID NOs: 80, 102, 139, 174, 199, and 221, respectively, SEQ ID NOs: 79, 110,

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively.

[0467] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of

SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC antibody moiety comprises heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 19 and 47, respectively, SEQ ID NOs: 20 and 48, respectively, SEQ ID NOs: 21 and 49, respectively, SEQ ID NOs: 22 and 50, respectively, SEQ ID NOs: 23 and 51, respectively, SEQ ID NOs: 24 and 52, respectively, SEQ ID NOs: 25 and 53, respectively, SEQ ID NOs: 26 and 54, respectively, SEQ ID NOs: 27 and 55, respectively, SEQ ID NOs: 28 and 56, respectively, SEQ ID NOs: 29 and 57, respectively, SEQ ID NOs: 30 and 58, respectively, SEQ ID NOs: 31 and 59, respectively, SEQ ID NOs: 32 and 60, respectively, SEQ ID NOs: 33 and 61, respectively, SEQ ID NOs: 34 and 62, respectively, SEQ ID NOs: 35 and 63, respectively, SEQ ID NOs: 36 and 64, respectively, SEQ ID NOs: 37 and 65, respectively, SEQ ID NOs: 38 and 66, respectively, SEQ ID NOs: 39 and 67, respectively, SEQ ID NOs: 40 and 68, respectively, SEQ ID NOs: 41 and 69, respectively, SEQ ID NOs: 42 and 70, respectively, SEQ ID NOs: 43 and 71, respectively, SEQ ID NOs: 44 and 72, respectively, SEQ ID NOs: 45 and 73, respectively, or SEQ ID NOs: 46 and 74, respectively.

[0468] In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the anti-RTMC CAR comprises an anti- RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC CAR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

[0469] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the HIV-1 RT peptide is HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9). In some embodiments, the MHC class I protein is HLA-A02. In some embodiments, the MHC class I protein is HLA-A*02:01. In some embodiments, the individual is human.

[0470] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT 181 (SEQ ID NO: 5), HIV-1 RT 181 M184V (SEQ ID NO: 6), HIV-1 RT 181 M184I (SEQ ID NO: 7), HIV-1 RT 181 Y181C (SEQ ID NO: 8), or HIV-1 RT 181 Y181C, M184V (SEQ ID NO: 9) peptide and HLA-A*02:01, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0471] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 (for example about any of 1,

3) amino acid substitutions, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human. [0472] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDRl comprising the amino acid sequence of SEQ ID NO: 240, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246; and ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0473] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising an HC-CDRl comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 (such as about any of 1, 2,

3, 4, or 5) amino acid substitutions; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0474] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a light chain variable domain sequence comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0475] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain sequence comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96; an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124; and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; and ii) a light chain variable domain sequence comprising an LC- CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189; an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207; and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0476] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising i) a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity, and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% (including for example at least about any of 96%, 97%, 98%, or 99%) sequence identity; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ

[0477] In some embodiments, there is provided a method of treating an HIV-1 infection in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-RTMC abTCR comprising a) an extracellular domain comprising an anti-RTMC antibody moiety that specifically binds to a complex comprising an HIV-1 RT peptide and an MHC class I protein comprising a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47- 74; b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence. In some embodiments, the individual is human.

[0478] For example, in some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-

NOs: 81, 103, 131, 169, 191, and 214, respectively, SEQ ID NOs: 80, 104, 132, 170, 195, and 215, respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82,

[0479] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

respectively, SEQ ID NOs: 76, 98, 133, 171, 196, and 216, respectively, SEQ ID NOs: 82, 105, 134, 164, 192, and 217, respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218,

[0480] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising HC-CDR1, HC-CDR2,

134, 164, 192, and 217, respectively, SEQ ID NOs: 83, 106, 135, 169, 191, and 218, respectively, SEQ ID NOs: 84, 107, 136, 172, 197, and 219, respectively, SEQ ID NOs: 85, 108,

respectively, or SEQ ID NOs: 96, 124, 163, 189, 207, and 239, respectively.

[0481] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of

[0482] In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 27 and 55, respectively. In some embodiments, the anti-RTMC abTCR comprises an anti- RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively, or variants thereof having, individually, at least about 95% (for example at least about any of 96%, 97%, 98%, or 99%) sequence identity. In some embodiments, the anti-RTMC abTCR comprises an anti-RTMC antibody moiety comprising heavy chain and light chain variable domains comprising the amino acid sequence of SEQ ID NOs: 30 and 58, respectively.

Methods for Diagnosis and Imaging Using anti-RTMC constructs

[0483] Labeled anti-RTMC antibody moieties and derivatives and analogs thereof, which specifically bind to an RTMC on the surface of a cell, can be used for diagnostic purposes to detect, diagnose, or monitor HIV-1 infection. For example, the anti-RTMC antibody moieties of the invention can be used in in situ, in vivo, ex vivo, and in vitro diagnostic assays or imaging assays.

[0484] Additional embodiments of the invention include methods of diagnosing HIV-1 infection in an individual (e.g., a mammal such as a human). The methods comprise detecting RTMC- presenting cells in the individual. In some embodiments, there is provided a method of diagnosing HIV-1 infection in an individual (e.g., a mammal, such as a human) comprising (a) administering an effective amount of a labeled anti-RTMC antibody moiety according to any of the embodiments described above to the individual; and (b) determining the level of the label in the individual, such that a level of the label above a threshold level indicates that the individual has the HIV-1 infection. The threshold level can be determined by various methods, including, for example, by detecting the label according to the method of diagnosing described above in a first set of individuals that have the HIV- 1 infection and a second set of individuals that do not have the HIV-1 infection, and setting the threshold to a level that allows for discrimination between the first and second sets. In some embodiments, the threshold level is zero, and the method comprises determining the presence or absence of the label in the individual. In some embodiments, the method further comprises waiting for a time interval following the

administering of step (a) to permit the labeled anti-RTMC antibody moiety to preferentially concentrate at sites in the individual where the RTMC is expressed (and for unbound labeled anti-RTMC antibody moiety to be cleared). In some embodiments, the method further comprises subtracting a background level of the label. Background level can be determined by various methods, including, for example, by detecting the label in the individual prior to administration of the labeled anti-RTMC antibody moiety, or by detecting the label according to the method of diagnosing described above in an individual that does not have the HIV-1 infection.

[0485] Anti-RTMC antibody moieties of the invention can be used to assay levels of RTMC- presenting cell in a biological sample using methods known to those of skill in the art. Suitable antibody labels are known in the art and include enzyme labels, such as, glucose oxidase;

radioisotopes, such as iodine ( 131 I, 125 I, 123 I, 121 I), carbon ( 14 C), sulfur ( 35 S), tritium ( 3 H), indium (^115mIn, ^113mIn, ¹¹²In, ^mIn), technetium (⁹⁹Tc, ^99mTc), thallium (²⁰¹Ti), gallium (⁶⁸Ga, ⁶⁷Ga), palladium (¹⁰³Pd), molybdenum (⁹⁹Mo), xenon (¹³³Xe), fluorine (¹⁸F), samarium (¹⁵³Sm), lutetium (¹⁷⁷Lu), gadolinium (¹⁵⁹Gd), promethium (¹⁴⁹Pm), lanthanum (¹⁴⁰La), ytterbium (¹⁷⁵Yb) , holmium (¹⁶⁶Ho), yttrium (⁹⁰Y), scandium (⁴⁷Sc), rhenium (¹⁸⁶Re, ¹⁸⁸Re), praseodymium (¹⁴²Pr), rhodium (¹⁰⁵Rh), and ruthenium (⁹⁷Ru); luminol; fluorescent labels, such as fluorescein and rhodamine; and bio tin. [0486] Techniques known in the art may be applied to labeled anti-RTMC antibody moieties of the invention. Such techniques include, but are not limited to, the use of bifunctional conjugating agents (see e.g., U.S. Pat. Nos. 5,756,065; 5,714,631; 5,696,239; 5,652,361; 5,505,931;

5,489,425; 5,435,990; 5,428,139; 5,342,604; 5,274,119; 4,994,560; and 5,808,003). Aside from the above assays, various in vivo and ex vivo assays are available to the skilled practitioner. For example, one can expose cells within the body of the subject to an anti-RTMC antibody moiety which is optionally labeled with a detectable label, e.g., a radioactive isotope, and binding of the anti-RTMC antibody moiety to the cells can be evaluated, e.g., by external scanning for radioactivity or by analyzing a sample {e.g., a biopsy or other biological sample) derived from a subject previously exposed to the anti-RTMC antibody moiety.

Articles of Manufacture and Kits

[0487] In some embodiments of the invention, there is provided an article of manufacture containing materials useful for the treatment of an HIV-1 infection, for delivering an anti-RTMC construct to a cell presenting an RTMC on its surface, or for isolation or detection of RTMC- presenting cells in an individual. The article of manufacture can comprise a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, etc. The containers may be formed from a variety of materials such as glass or plastic. Generally, the container holds a composition which is effective for treating an HIV-1 infection, and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is an anti-RTMC construct of the invention. The label or package insert indicates that the composition is used for treating the particular condition. The label or package insert will further comprise instructions for administering the anti-RTMC construct composition to the patient. Articles of manufacture and kits comprising combinatorial therapies described herein are also contemplated.

[0488] Package insert refers to instructions customarily included in commercial packages of therapeutic products that contain information about the indications, usage, dosage,

administration, contraindications and/or warnings concerning the use of such therapeutic products. In some embodiments, the package insert indicates that the composition is used for treating HIV- 1 infection.

[0489] Additionally, the article of manufacture may further comprise a second container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

[0490] Kits are also provided that are useful for various purposes, e.g., for treatment of an HIV- 1 infection, for delivering an anti-RTMC construct to a cell presenting an RTMC on its surface, or for isolation or detection of RTMC-presenting cells in an individual, optionally in

combination with the articles of manufacture. Kits of the invention include one or more containers comprising an anti-RTMC construct composition (or unit dosage form and/or article of manufacture), and in some embodiments, further comprise another agent (such as the agents described herein) and/or instructions for use in accordance with any of the methods described herein. The kit may further comprise a description of selection of individuals suitable for treatment. Instructions supplied in the kits of the invention are typically written instructions on a label or package insert (e.g. , a paper sheet included in the kit), but machine-readable instructions (e.g., instructions carried on a magnetic or optical storage disk) are also acceptable.

[0491] For example, in some embodiments, the kit comprises a composition comprising an anti- RTMC construct (e.g., a full-length anti-RTMC antibody, a multi- specific anti-RTMC molecule (such as a bispecific anti-RTMC antibody), or an anti-RTMC immunoconjugate). In some embodiments, the kit comprises a) a composition comprising an anti-RTMC construct, and b) an effective amount of at least one other agent, wherein the other agent increases the expression of MHC class I proteins and/or enhances the surface presentation of HIV- 1 RT peptides by MHC class I proteins (e.g. , ΙΡΝγ, IFNP, IFNa, or Hsp90 inhibitor). In some embodiments, the kit comprises a) a composition comprising an anti-RTMC construct, and b) instructions for administering the anti-RTMC construct composition to an individual for treatment of an HIV- 1 infection. In some embodiments, the kit comprises a) a composition comprising an anti-RTMC construct, b) an effective amount of at least one other agent, wherein the other agent increases the expression of MHC class I proteins and/or enhances the surface presentation of HIV- 1 RT peptides by MHC class I proteins (e.g. , ΙΡΝγ, IFNP, IFNa, or Hsp90 inhibitor), and c) instructions for administering the anti-RTMC construct composition and the other agent(s) to an individual for treatment of an HIV- 1 infection. The anti-RTMC construct and the other agent(s) can be present in separate containers or in a single container. For example, the kit may comprise one distinct composition or two or more compositions wherein one composition comprises an anti-RTMC construct and another composition comprises another agent. [0492] In some embodiments, the kit comprises a) a composition comprising an anti-RTMC construct (e.g., a full-length anti-RTMC antibody, a multi- specific anti-RTMC molecule (such as a bispecific anti-RTMC antibody), or an anti-RTMC immunoconjugate), and b) instructions for combining the anti-RTMC construct with cells (such as cells, e.g. , immune cells, derived from an individual) to form a composition comprising anti-RTMC construct/cell conjugates and administering the anti-RTMC construct/cell conjugate composition to the individual for treatment of an HIV- 1 infection. In some embodiments, the kit comprises a) a composition comprising an anti-RTMC construct, and b) a cell (such as a cytotoxic cell). In some

embodiments, the kit comprises a) a composition comprising an anti-RTMC construct, b) a cell (such as a cytotoxic cell), and c) instructions for combining the anti-RTMC construct with the cell to form a composition comprising anti-RTMC construct/cell conjugates and administering the anti-RTMC construct/cell conjugate composition to an individual for the treatment of an HIV- 1 infection. In some embodiments, the kit comprises a composition comprising an anti- RTMC construct in association with a cell (such as a cytotoxic cell). In some embodiments, the kit comprises a) a composition comprising an anti-RTMC construct in association with a cell (such as a cytotoxic cell), and b) instructions for administering the composition to an individual for the treatment of an HIV- 1 infection. In some embodiments, the association is by conjugation of the anti-RTMC construct to a molecule on the surface of the cell. In some embodiments, the association is by insertion of a portion of the anti-RTMC construct into the outer membrane of the cell.

[0493] In some embodiments, the kit comprises a nucleic acid (or set of nucleic acids) encoding an anti-RTMC construct (e.g. , a full-length anti-RTMC antibody, a multi- specific anti-RTMC molecule (such as a bispecific anti-RTMC antibody), an anti-RTMC CAR, an anti-RTMC abTCR, or an anti-RTMC immunoconjugate) or polypeptide portions thereof. In some embodiments, the kit comprises a) a nucleic acid (or set of nucleic acids) encoding an anti- RTMC construct or polypeptide portions thereof, and b) a host cell (such as an effector cell) for expressing the nucleic acid (or set of nucleic acids). In some embodiments, the kit comprises a) a nucleic acid (or set of nucleic acids) encoding an anti-RTMC construct or polypeptide portions thereof, and b) instructions for i) expressing the anti-RTMC construct in a host cell (such as an effector cell, e.g. , a T cell), ii) preparing a composition comprising the anti-RTMC construct or the host cell expressing the anti-RTMC construct, and iii) administering the composition comprising the anti-RTMC construct or the host cell expressing the anti-RTMC construct to an individual for the treatment of an HIV- 1 infection. In some embodiments, the host cell is derived from the individual. In some embodiments, the kit comprises a) a nucleic acid (or set of nucleic acids) encoding an anti-RTMC construct or polypeptide portions thereof, b) a host cell (such as an effector cell) for expressing the nucleic acid (or set of nucleic acids), and c) instructions for i) expressing the anti-RTMC construct in the host cell, ii) preparing a composition comprising the anti-RTMC construct or the host cell expressing the anti-RTMC construct, and iii) administering the composition comprising the anti-RTMC construct or the host cell expressing the anti-RTMC construct to an individual for the treatment of an HIV- 1 infection.

[0494] In some embodiments, the kit comprises a nucleic acid encoding an anti-RTMC CAR or anti-RTMC abTCR. In some embodiments, the kit comprises a vector comprising a nucleic acid encoding an anti-RTMC CAR or anti-RTMC abTCR. In some embodiments, the kit comprises a) a vector comprising a nucleic acid encoding an anti-RTMC CAR or anti-RTMC abTCR, and b) instructions for i) introducing the vector into effector cells, such as T cells derived from an individual, ii) preparing a composition comprising the anti-RTMC CAR or anti-RTMC abTCR effector cells, and iii) administering the anti-RTMC CAR or anti-RTMC abTCR effector cell composition to the individual for treatment of an HIV- 1 infection.

[0495] The kits of the invention are in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g. , sealed Mylar or plastic bags), and the like. Kits may optionally provide additional components such as buffers and interpretative information. The present application thus also provides articles of manufacture, which include vials (such as sealed vials), bottles, jars, flexible packaging, and the like.

[0496] The instructions relating to the use of the anti-RTMC construct compositions generally include information as to dosage, dosing schedule, and route of administration for the intended treatment. The containers may be unit doses, bulk packages (e.g. , multi-dose packages) or sub- unit doses. For example, kits may be provided that contain sufficient dosages of an anti-RTMC construct (e.g., a full-length anti-RTMC antibody, a multi- specific anti-RTMC molecule (such as a bispecific anti-RTMC antibody), an anti-RTMC CAR, an anti-RTMC abTCR, or an anti- RTMC immunoconjugate) as disclosed herein to provide effective treatment of an individual for an extended period, such as any of a week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, or more. Kits may also include multiple unit doses of the anti-RTMC construct and pharmaceutical compositions and instructions for use and packaged in quantities sufficient for storage and use in pharmacies, for example, hospital pharmacies and compounding pharmacies. EXEMPLARY EMBODIMENTS

[0497] Embodiment 1. An isolated anti-RTMC construct comprising an antibody moiety that specifically binds to a complex comprising a human immunodeficiency virus 1 (HIV-1) reverse transcriptase (RT) peptide and a major histocompatibility (MHC) class I protein ("RTMC").

[0498] Embodiment 2. The isolated anti-RTMC construct of embodiment 1, wherein the HIV-1 RT/MHC class I complex is present on a cell surface.

[0499] Embodiment 3. The isolated anti-RTMC construct of embodiment 1, wherein the HIV-1 RT/MHC class I complex is present on the surface of a T cell.

[0500] Embodiment 4. The isolated anti-RTMC construct of any one of embodiments 1-3, wherein the MHC class I protein is human leukocyte antigen (HLA)-A.

[0501] Embodiment 5. The isolated anti-RTMC construct of embodiment 4, wherein the MHC class I protein is HLA-A02.

[0502] Embodiment 6. The isolated anti-RTMC construct of embodiment 5, wherein the MHC class I protein is selected from the group consisting of HLA-A*02:01, HLA-A*02:02, HLA- A*02:06, HLA-A*02:07, and HLA-A*02: 11.

[0503] Embodiment 7. The isolated anti-RTMC construct of embodiment 6, wherein the MHC class I protein is HLA-A*02:01.

[0504] Embodiment 8. The isolated anti-RTMC construct of any one of embodiments 1-7, wherein the antibody moiety cross-reacts with a complex comprising the HIV-1 RT peptide and a second MHC class I protein having a different HLA allele than the MHC class I protein.

[0505] Embodiment 9. The isolated anti-RTMC construct of any one of embodiments 1-8, wherein the HIV-1 RT peptide is 8 to 12 amino acids in length.

[0506] Embodiment 10. The isolated anti-RTMC construct of any one of embodiments 1-9, wherein the HIV-1 RT peptide is derived from the region corresponding to amino acids 181-189 of SEQ ID NO: 1.

[0507] Embodiment 11. The isolated anti-RTMC construct of any one of embodiments 1-10, wherein the HIV-1 RT peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-18.

[0508] Embodiment 12. The isolated anti-RTMC construct of embodiment 10, wherein the HIV- 1 RT peptide has the amino acid sequence of YQYVDDLYV (SEQ ID NO: 6). [0509] Embodiment 13. The isolated anti-RTMC construct of embodiment 12, wherein the isolated anti-RTMC construct cross-reacts with a complex comprising a variant of the HIV-1 RT peptide having the amino acid sequence of any one of YQYMDDLYV (SEQ ID NO: 5), YQYIDDLYV (SEQ ID NO: 7), CQYMDDLYV (SEQ ID NO: 8), or CQYVDDLYV (SEQ ID NO: 9) and the MHC class I protein.

[0510] Embodiment 14. The isolated anti-RTMC construct of embodiment 10, wherein the HIV- 1 RT peptide has the amino acid sequence of YQYMDDLYV (SEQ ID NO: 5).

[0511] Embodiment 15. The isolated anti-RTMC construct of embodiment 14, wherein the isolated anti-RTMC construct cross-reacts with a complex comprising a variant of the HIV-1 RT peptide having the amino acid sequence of any one of YQYVDDLYV (SEQ ID NO: 6), YQYIDDLYV (SEQ ID NO: 7), CQYMDDLYV (SEQ ID NO: 8), or CQYVDDLYV (SEQ ID NO: 9) and the MHC class I protein.

[0512] Embodiment 16. The isolated anti-RTMC construct of any one of embodiments 1-15, wherein the antibody moiety is human, humanized, synthetic, or semi- synthetic.

[0513] Embodiment 17. The isolated anti-RTMC construct of any one of embodiments 1-16, wherein the antibody moiety is a full-length antibody, a Fab, a Fab', a (Fab')2, an Fv, or a single chain Fv (scFv).

[0514] Embodiment 18. The isolated anti-RTMC construct of any one of embodiments 1-17, wherein the antibody moiety binds to the HIV-1 RT/MHC class I complex with an equilibrium dissociation constant (Kd) from about 0.1 pM to about 500 nM.

[0515] Embodiment 19. The isolated anti-RTMC construct of any one of embodiments 1-18, wherein the isolated anti-RTMC construct binds to the HIV-1 RT/MHC class I complex with a Kd from about 0.1 pM to about 500 nM.

[0516] Embodiment 20. The isolated anti-RTMC construct of any one of embodiments 1-19, wherein the antibody moiety comprises:

i) a heavy chain variable domain comprising a heavy chain complementarity determining region (HC-CDR) 1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 amino acid substitutions; and ii) a light chain variable domain comprising a light chain complementarity determining region (LC-CDR) 1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 amino acid substitutions.

[0517] Embodiment 21. The isolated anti-RTMC construct of any one of embodiments 1-19, wherein the antibody moiety comprises:

i) a heavy chain variable domain comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, or a variant thereof comprising up to about 5 amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, or a variant thereof comprising up to about 5 amino acid substitutions, and an HC- CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 amino acid substitutions; and

ii) a light chain variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, or a variant thereof comprising up to about 5 amino acid substitutions, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, or a variant thereof comprising up to about 3 amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 amino acid substitutions.

[0518] Embodiment 22. The isolated anti-RTMC construct of any one of embodiments 1-19, wherein the antibody moiety comprises:

i) a heavy chain (HC) variable domain comprising an HC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 75-96, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 97-124, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 125-163; or a variant thereof comprising up to about 5 amino acid substitutions in the HC-CDR regions; and

ii) a light chain (LC) variable domain comprising an LC-CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 164-189, an LC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 190-207, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 208-239; or a variant thereof comprising up to about 5 amino acid substitutions in the LC-CDR regions.

[0519] Embodiment 23. The isolated anti-RTMC construct of embodiment 21 or 22, wherein the antibody moiety comprises a) a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% sequence identify to any one of SEQ ID NOs: 19-46; and b) a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% sequence identity to any one of SEQ ID NOs: 47-74.

[0520] Embodiment 24. The isolated anti-RTMC construct of embodiment 23, wherein the antibody moiety comprises a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46 and a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74.

[0521] Embodiment 25. The isolated anti-RTMC construct of any one of embodiments 1-24, wherein the isolated anti-RTMC construct is a full-length antibody.

[0522] Embodiment 26. The isolated anti-RTMC construct of any one of embodiments 1-25, wherein the isolated anti-RTMC construct is monospecific.

[0523] Embodiment 27. The isolated anti-RTMC construct of any one of embodiments 1-25, wherein the isolated anti-RTMC construct is multispecific.

[0524] Embodiment 28. The isolated anti-RTMC construct of embodiment 27, wherein the isolated anti-RTMC construct is bispecific.

[0525] Embodiment 29. The isolated anti-RTMC construct of embodiment 27 or 28, wherein the isolated anti-RTMC construct is a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into- hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a heteromultimeric antibody, or a heteroconjugate antibody.

[0526] Embodiment 30. The isolated anti-RTMC construct of embodiment 29, wherein the isolated anti-RTMC construct is a tandem scFv comprising two scFvs linked by a peptide linker.

[0527] Embodiment 31. The isolated anti-RTMC construct of embodiment 30, wherein the peptide linker comprises the amino acid sequence of SEQ ID NO: 276.

[0528] Embodiment 32. The isolated anti-RTMC construct of any one of embodiments 27-31, wherein the isolated anti-RTMC construct further comprises a second antibody moiety that specifically binds to a second antigen.

[0529] Embodiment 33. The isolated anti-RTMC construct of embodiment 32, wherein the second antigen is an antigen on the surface of a T cell.

[0530] Embodiment 34. The isolated anti-RTMC construct of embodiment 33, wherein wherein the second antigen is selected from the group consisting of CD3y, CD35, CD3s, CD3ζ, CD28,

OX40, GITR, CD137, CD27, CD40L and HVEM. [0531] Embodiment 35. The isolated anti-RTMC construct of embodiment 33, wherein the second antigen is CD3s, and wherein the isolated anti-RTMC construct is a tandem scFv comprising an N-terminal scFv specific for the HIV-1 RT/MHC class I complex and a C- terminal scFv specific for CD3s.

[0532] Embodiment 36. The isolated anti-RTMC construct of embodiment 33, wherein the T cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, and a natural killer T cell.

[0533] Embodiment 37. The isolated anti-RTMC construct of embodiment 32, wherein the second antigen is an antigen on the surface of a natural killer cell, a neutrophil, a monocyte, a macrophage or a dendritic cell.

[0534] Embodiment 38. The isolated anti-RTMC construct of any one of embodiments 1-24, wherein the isolated anti-RTMC construct is a chimeric antigen receptor (CAR).

[0535] Embodiment 39. The isolated anti-RTMC construct of embodiment 38, wherein the chimeric antigen receptor comprises an extracellular domain comprising the antibody moiety, a transmembrane domain, and an intracellular signaling domain comprising a CD3ζ intracellular signaling sequence and a CD28 and/or 4- IBB intracellular signaling sequence.

[0536] Embodiment 40. The isolated anti-RTMC construct of any one of embodiments 1-24, wherin the isolated anti-RTMC construct is a chimeric antibody/T cell receptor (abTCR) comprising an extracellular domain comprising the antibody moiety and a T cell receptor (TCR) module (TCRM) comprising TCR transmembrane domains.

[0537] Embodiment 41. The isolated anti-RTMC construct of embodiment 40, wherein the TCRM is capable of recruiting at least one TCR-associated signaling module.

[0538] Embodiment 42. The isolated anti-RTMC construct of embodiment 41, wherein the TCR-associated signaling module is selected from the group consisting of CD35s, CD3y8, and ζζ·

[0539] Embodiment 43. The isolated anti-RTMC construct of any one of embodiments 40-42, wherein the antibody moiety comprises:

a) a first polypeptide chain comprising a first antigen-binding domain comprising V_H and C_HI antibody domains; and

b) a second polypeptide chain comprising a second antigen-binding domain comprising V_L and C_L antibody domains, wherein the V_H and C_HI domains of the first antigen-binding domain and the V_L and C_L domains of the second antigen-binding domain form a Fab-like antigen-binding module that specifically binds to the RTMC.

[0540] Embodiment 44. The isolated anti-RTMC construct of any one of embodiments 1-24, wherein the isolated anti-RTMC construct is an immunoconjugate comprising the antibody moiety and an effector molecule.

[0541] Embodiment 45. The isolated anti-RTMC construct of embodiment 44, wherein the effector molecule is a therapeutic agent selected from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid.

[0542] Embodiment 46. The isolated anti-RTMC construct of embodiment 45, wherein the therapeutic agent is a drug or a toxin.

[0543] Embodiment 47. The isolated anti-RTMC construct of embodiment 44, wherein the effector molecule is a label.

[0544] Embodiment 48. A host cell expressing the isolated anti-RTMC construct of any one of embodiments 1-47.

[0545] Embodiment 49. A nucleic acid encoding one or more polypeptides contained in the isolated anti-RTMC construct of any one of embodiments 1-47.

[0546] Embodiment 50. A vector comprising the nucleic acid of embodiment 49.

[0547] Embodiment 51. An effector cell expressing the isolated anti-RTMC construct of any one of embodiments 38-43.

[0548] Embodiment 52. The effector cell of embodiment 51, wherein the effector cell is a T cell.

[0549] Embodiment 53. A pharmaceutical composition comprising the isolated anti-RTMC construct of any one of embodiments 1-46, the nucleic acid of embodiment 49, the vector of embodiment 50, or the effector cell of embodiment 51 or 52.

[0550] Embodiment 54. A method for detecting a cell presenting a complex comprising an HIV- 1 RT peptide and an MHC class I protein on its surface, comprising contacting the cell with the isolated anti-RTMC construct of embodiment 47 and detecting the presence of the label on the cell.

[0551] Embodiment 55. A method for treating an individual having an HIV-1 infection, comprising administering to the individual an effective amount of the pharmaceutical composition of embodiment 53. [0552] Embodiment 56. A method for treating an individual having an HIV-1 infection, comprising administering to the individual an effective amount of the effector cell of embodiment 51 or 52.

[0553] Embodiment 57. A method of diagnosing an individual having an HIV-1 infection, comprising:

a) administering an effective amount of the isolated anti-RTMC construct of embodiment 47 to the individual; and

b) determining the level of the label in the individual, wherein a level of the label above a threshold level indicates that the individual has the HIV-1 infection.

[0554] Embodiment 58. A method of diagnosing an individual having an HIV-1 infection, comprising:

a) contacting a sample derived from the individual with the isolated anti-RTMC construct of embodiment 47; and

b) determining the number of cells bound with the isolated anti-RTMC construct in the sample, wherein a value for the number of cells bound with the isolated anti-RTMC construct above a threshold level indicates that the individual has the HIV-1 infection.

[0555] Embodiment 59. The method of any one of embodiments 55-58, wherein the individual is human.

[0556] Embodiment 60. The isolated anti-RTMC construct of any one of embodiments 1-19, wherein the HIV-1 RT peptide is HIV-1 RT 181, wherein the antibody moiety comprises HC- CDR and LC-CDR sequences, and wherein a bispecific antibody according to embodiment 35 and comprising the HC-CDR and LC-CDR sequences of the isolated anti-RTMC construct is stable in an aqueous formulation for at least about 2 years.

[0557] Embodiment 61. The isolated anti-RTMC construct of embodiment 60, wherein the aqueous formulation is stored at about 4° C.

[0558] Embodiment 62. The isolated anti-RTMC construct of embodiment 60 or 61, wherein the bispecific antibody in the aqueous formulation retains at least about 40% of its target cell-killing activity for at least about 2 years.

[0559] Embodiment 63. The isolated anti-RTMC construct of any one of embodiments 60-62, wherein the anti-RTMC construct is a bispecific antibody according to embodiment 35. EXAMPLES

[0560] Those skilled in the art will recognize that several embodiments are possible within the scope and spirit of this invention. The invention will now be described in greater detail by reference to the following non-limiting examples. The following examples further illustrate the invention but, of course, should not be construed as in any way limiting its scope.

Materials

Cell Samples, Cell Lines, and Antibodies

[0561] The cell lines include: liver adenocarcinoma cell line SK-HEP-1 (ATCC HTB-52; HLA- A2⁺, HIV-l^") and lymphoblast cell line T2 (ATCC CRL-1992; HLA-A2⁺, HIV-l^"). T2 is a TAP- deficient cell line. The cell lines were cultured in RPMI 1640 supplemented with 5% FCS, penicillin, streptomycin, 2 mmol/L glutamine, and 2-mercaptoethanol at 37° C/5% C0₂.

[0562] All peptides were purchased and synthesized by Genemed Synthesis, Inc. (San Antonio, Tex.) or Elim Biopharm (Hayward, CA). Peptides were >90% pure. The peptides were dissolved in DMSO at 10 mg/mL and frozen at -80 °C. Recombinant HIV-l RT peptide/HLA-A*02:01 and control peptide/HLA-A*02:01 complexes were prepared by refolding the peptides with recombinant HLA-A02 (biotinylated at the C-terminus using BirA, Bio tin- Protein ligase from Avidity (Aurora, Colorado)) and beta-2 microglobulin (β2Μ) (~2M). 20 control peptides (P20, SEQ ID NOs: 255-274) that bind HLA-A*02:01 were generated from the following 15 genes: BCR, BTG2, CALR, CD247, CSF2RA, CTSG, DDX5, DMTN, HLA-E, IFI30, IL7, PIM1, PPP2R1B, RPS6KB 1, SSR1, and β-globin.

Example 1. Production of biotinylated HIV-l RT/HLA-A*02:01 complex monomer

[0563] Biotinylated HIV-l RT/HLA-A*02:01 complex monomers were prepared according to standard protocols (John D. Altman and Mark M. Davis, Current Protocols in Immunology 17.3.1-17.3.33, 2003). In brief, DNA encoding full-length human beta-2 microglobulin (p2m) was synthesized by Genewiz and cloned into vector pET-27b. DNA encoding HLA-A*02:01 ECD-BSP (BirA substrate peptide (BSP) fused to the C-terminus of HLA-A*02:01 extracellular domain (ECD)) was also synthesized by Genewiz and cloned into vector pET-27b. The vectors expressing human β2ιη and HLA-A*02:01 ECD-BSP were transformed into E.coli BL21 cells separately, and expressed proteins were isolated as inclusion bodies from bacterial culture. Peptide ligand HIV-l RT peptide 181 M184V (YQYVDDLYV, SEQ ID NO: 6) was refolded with human p2m and HLA-A*02:01 ECD-BSP to form HIV-1 RT peptide/HLA-A*02:01 complex monomer. Folded peptide/HLA-A*02:01 monomers were concentrated by

ultrafiltration and further purified through size-exclusion chromatography. HiPrep 26/60

Sephacryl S-300 HR was equilibrated with 1.5 column volumes of Hyclone Dulbecco's

Phosphate Buffered Saline solution (Thermo Scientific, Cat No. SH3002802). The unpurified sample was loaded and eluted with 1 column volume. The first peak, corresponding to misfolded aggregates, eluted at approximately 111 mL, the peak corresponding to the properly folded MHC complex was observed at 212 mL, and the peak corresponding to free β2Μ was observed at 267 mL (data not shown). Peptide/HLA-A*02:01 monomers were biotinylated via BirA- mediated enzymatic reaction. Biotinylated peptide/HLA-A*02:01 monomers were stored in PBS at -80°C.

[0564] SDS-PAGE of the purified HIV-1 RT peptide/MHC complex can be performed to determine protein purity. For example, ^g of the protein complex is mixed with 2.5μί of the NuPAGE LDS Sample Buffer (Life Technologies, NP0008) and brought up to ΙΟμί with deionized water. The sample is heated at 70°C for 10 minutes, and then loaded onto the gel. Gel electrophoresis is performed at 180V for 1 hour.

Example 2. Selection and Characterization of scFv Specific for HIV-1 RT/HLA-A*02:01 Complexes.

[0565] A collection of human scFv antibody phage display libraries (diversity = lOxlO¹⁰) constructed by Eureka Therapeutics was used for the selection of human mAbs specific to HIV-

1 RT 181 M184V/HLA-A*02:01/p2M complexes. 24 fully human phage scFv libraries were used to pan against HIV-1 RT/HLA-A*02:01 complex. In order to reduce the conformational change of MHCI complex introduced by immobilizing the protein complex onto plastic surfaces, solution panning and cell panning were used in place of conventional plate panning. In solution panning, recombinant biotinylated antigens were first mixed with the human scFv phage library after extended washing with PBS buffer, and then antigen-scFv antibody phage complexes were pulled down by streptavidin-conjugated Dynabeads M-280 through a magnetic rack. The bound clones were then eluted and used to infect E.coli XLl-Blue cells. In cell panning, T2 cells loaded with HIV- 1 RT peptide were first mixed with the human scFv phage library. T2 cells are a TAP- deficient, HLA-A*02:01⁺ lymphoblast cell line. To load peptide, T2 cells were pulsed with peptides (50μg/ml) in serum-free RPMI1640 medium. After extended washing with PBS, peptide- loaded T2 cells with bound scFv antibody phage were spun down. The bound clones were then eluted and used to infect E.coli XLl-Blue cells. The phage clones expressed in bacteria were then purified. The panning was performed for 3 rounds with either solution panning, cell panning or a combination of solution and cell panning to enrich for scFv phage clones that bound HIV-1 RT/HLA-A*02:01 specifically, and 1350 clones were selected for screening.

[0566] Streptavidin ELISA plates were coated with monomeric biotinylated HIV-1 RT 181 peptide/HLA-A*02:01 complex monomer or monomeric biotinylated C3 control peptide/HLA-

A*02:01 monomer respectively. C3 peptide is human nuclear protein p68-derived peptide

YLLPAIVHI (SEQ ID NO: 255). Individual phage clones from enriched phage display panning pools against HIV-1 RT 181 peptide/HLA-A*02:01 complex were incubated in the coated plates. Binding of the phage clones was detected by HRP-conjugated anti-M13 antibodies and developed using HRP substrate. The absorbance was read at 450nm. 214 positive clones were identified through ELISA screening of the 1350 phage clones enriched from phage panning. 39 unique clones were identified by DNA sequencing of the 214 ELISA-positive phage clones.

[0567] Specific and unique clones were further tested for their binding to HLA-A*02:01/peptide complexes on the cell surface by flow cytometry (FACS analysis) using HIV-1 RT-loaded live

T2 cells. T2 cells were loaded with HIV-1 RT 181 WT peptide, HIV-1 RT 181 M184V peptide, or a control mixture of 19 peptides (P19, SEQ ID NOs: 255-273). Controls included T2 cells without peptide loading (T2), R-PE conjugated horse anti-mouse IgG control (secondary antibody only), and cells stained with a negative control phage (NC phage). In brief, peptide- loaded T2 cells were first stained with purified scFv phage clones, followed by a second staining with mouse anti-M13 mAb, and a third staining with R-PE conjugated horse anti- mouse IgG from Vector Labs. Each staining step was performed for between 30-60 minutes on ice and the cells were washed twice between staining s. As reported in Table 7, among the 39 clones tested, all 39 recognized HIV-1 RT 181 M184V-loaded T2 cells specifically, and 37 also recognized

HIV-1 RT 181 WT-loaded T2 cells specifically. These phage clones specifically bound to HIV-1

RT-loaded T2 cells and did not recognize T2 cells loaded with control peptide mixture P19 in the context of HLA-A*02:01, or T2 cells without peptide loaded. To confirm peptide loading,

T2-P19 and T2-RT 181 cells were stained with BB7.2, an anti-HLA-A*02 specific antibody.

Peptide binding to MHC complex stabilizes cell- surface MHC complexes and this change in stability can be detected with BB7.2. Therefore, T2 cells loaded with MHC-binding peptide and stained with BB7.2 will have an enhanced fluorescence signal compared to T2 cells without peptide loading. BB7.2 binding data showed the HIV-1 RT peptides and control peptide mixture P19 were able to bind HLA-A*02:01 molecules and form surface peptide/MHC complexes as seen in the change in MFI compared to T2 cells.

Table 7

Anti-HIV-1 RT Clone 37 176 8.34 11.4 10.7

Anti-HIV-1 RT Clone 38 491 434 54.7 9.98

Anti-HIV-1 RT Clone 39 710 543 18.7 6.78

Anti-HIV-1 RT Clone 40 580 583 299 62.2

Anti-HIV-1 RT Clone 41 549 547 63.1 7.74

[0568] On average, each nucleated cell in the human body expresses about half a million different peptide/MHC Class I complexes. In order to develop anti-peptide/MHCI-complex antibodies into drugs with high specificity and therapeutic index, it is essential for the antibodies to specifically recognize the target peptide/MHCI complex, but not the MHCI molecule itself, or MHCI molecules bound to other peptides presented on cell surfaces. The phage clones were screened against a mixture of 19 endogenous HLA-A*02:01 peptides, which were derived from proteins normally expressed in multiple types of nucleated human cells, such as globin alpha chain, beta chain, nuclear protein p68, and the like. As reported in Table 7, the HIV-1 RT peptide/HLA-A*02:01-specific antibody phage clones bound HIV-1 RT peptide/HLA-A*02:01 complex, but not HLA-A*02:01 complexes folded with endogenous peptides. We conclude that the identified antibodies are specific to HIV-1 RT peptide/HLA-A*02:01 complexes, and do not recognize HLA-A*02:01 molecules bound to other HLA-A*02:01 -restricted peptides.

Example 3. Characterization of FACS-positive HIV-1 RT-specific phage clones

Cross-reactivity to HIV-1 RT peptide 181 variants

[0569] The HIV-1 RT 181 peptide selected in this invention is highly conserved among various HIV-1 strains. Clones selected from FACS binding analysis against WT and M184V HIV-1 RT peptide 181-loaded T2 cells were characterized further for cross-reactivity towards HIV-1 RT peptide 181 variant/HLA-A*02:01 complexes on live cell surfaces by FACS analysis using variant HIV-1 RT peptide- loaded T2 cells. The variant peptides differed from the WT HIV-1 RT 181 peptide by one or two amino acids at positions 181 and/or 184. The variant peptide sequences tested included HIV-1 RT 181 M184V (YQYVDDLYV, SEQ ID NO: 6), HIV-1 RT 181 M184I (YQYIDDLYV, SEQ ID NO: 7), HIV-1 RT 181 Y181C (CQYMDDLYV, SEQ ID NO: 8), and HIV-1 RT 181 Y181C/M184V (CQYVDDLYV, SEQ ID NO: 9).

[0570] In brief, T2 cells were loaded with the HIV-1 RT 181 peptide. Controls included T2 cells without peptide loading (T2). The peptide- loaded T2 cells were stained with purified scFv phage clones, followed by a second staining with a mouse anti-M13 mAb and a third staining with R- PE conjugated horse anti-mouse IgG from Vector Labs. Each staining step was performed on ice for 30-60 minutes and the cells were washed twice between each staining. As reported in Table 8, among the 8 clones tested, 6 recognized all HIV-1 RT 181 peptide- loaded T2 cells specifically, while 2 were unable to bind to HIV-1 RT WT-loaded T2 cells, 1 of which was also unable to bind to T2 cells loaded with HIV-1 RT 181 Y181C or HIV-1 RT 181 Y181C/M184V. BB7.2 binding data indicate that all of the HIV-1 RT peptides were able to bind HLA-A*02:01 molecules to form surface peptide/MHC complexes, though T2 cells loaded with HIV-1 RT 181 Y181C or HIV-1 RT 181 Y181C/M184V showed much higher surface expression than T2 cells loaded with the other HIV-1 RT peptides (FIG. 1).

Table 8

Epitope mapping by alanine walking

[0571] To investigate with precision the epitope for the mAb recognition, HIV-1 RT peptides with alanine substitutions at positions 1, 2, 3, 4, 5, 6, 7, 8, or 9 (Table 9) were pulsed onto the surface of T2 cells. Antibody phage clones were then tested for binding to these peptide- loaded T2 cells by FACS analysis. Mean fluorescent intensity (MFI) values of each staining are shown in Table 9. All the antibody phage clones tested recognized the small conformational epitope formed by the HIV-1 RT peptide and its surrounding MHC alpha chain residues, and the key peptide residues interacting with the various antibodies appeared to reside in the C-terminal half of the peptide, as most clones were sensitive to substitution at positions 6 and 7 (Table 10). Positions 5, 8, and 9 appeared to affect peptide loading based on BB7.2 binding (FIG. 1).

Controls included T2 cells without peptide loading ("T2" in Table 10 and "T2 Cell + 2^nd Antibody in FIG. 1).

Table 9

Table 10

Antibody binding specificity evaluation against endogenous peptide [0572] Phage clones may also be screened against individual endogenous HLA-A*02:01 peptides derived from proteins normally expressed in multiple types of nucleated human cells, such as the peptides contained in P20 (SEQ ID NOs: 255-274). For example, recombinant peptide/HLA-A*02:01 complexes folded with 20 endogenous peptides (SEQ ID NOs: 255-274) separately are coated on streptavidin plates and antibody binding is determined through ELISA analysis. In brief, individual phage clones are incubated on the peptide/HLA-A*02:01 complex- coated plates. Binding of the phage clones is detected by HRP-conjugated anti-M13 antibodies and developed using HRP substrate. The absorbance is read at 450nm.

Example 4. Engineering bispecific antibodies

[0573] Bispecific antibodies (BsAbs) were generated using scFv sequences of the HIV-1 RT/HLA-A*02:01-specific phage clones. The BsAbs are single-chain bispecific antibodies comprising the scFv sequence of an HIV-1 RT/HLA-A*02:01-specific phage clone (from N- terminus to C-terminus: light chain variable region (LCVR), scFv linker (SEQ ID NO: 276), heavy chain variable region (HCVR)) fused to an anti-human CD3s mouse monoclonal scFv at the C-terminal end (SEQ ID NO: 277) (Brischwein, K. et al., Molecular Immunology 43: 1129- 1143, 2006) by a BsAb linker (SEQ ID NO: 278). The phage clone heavy and light chain variable regions used for generating the BsAbs are listed in Table 11. DNA fragments coding for the HIV-1 RT scFv and the anti- human CD3s scFv were synthesized by Genewiz and subcloned into Eureka's mammalian expression vector pGSN-Hyg using standard DNA techniques. A hexhistamine tag was inserted at the C-terminal end for antibody purification and detection. Chinese hamster ovary (CHO) cells were transfected with the BsAb expression vector, and then cultured for 7 days for BsAb antibody production. CHO cell supernatants containing secreted HIV-1 RT BsAb molecules were collected. BsAbs were purified by affinity chromatography using a HisTrap HP column (GE healthcare) and an AKTA FPLC system. Briefly, CHO cell culture was clarified and loaded onto the column with low imidazole concentration (20 mM), and then an isocratic high imidazole concentration elution buffer (500 mM) was used to elute the bound BsAb proteins. Purity and molecular weight of the purified HIV-1 RT BsAbs can be determined under reducing conditions by gel electrophoresis. For example, 4μg of the protein is mixed with 2.5μί of the NuPAGE LDS Sample Buffer (Life Technologies, NP0008) and brought up to ΙΟμί with deionized water. The sample is heated at 70°C for 10 minutes, and then loaded onto the gel. Gel electrophoresis is performed at 180V for 1 hour. Table 11

[0574] Antibody aggregation can be assessed by size-exclusion chromatography (SEC). For example, 50μί of the sample is injected into a SEC column (for example Agilent, BioSEC- 3,300A, 4.6x300mm) while flowing a buffer consisting of Dulbecco' s Phosphate Buffered Saline (Fisher Scientific, SH30028.FS) and 0.2M arginine adjusted to pH 7.0. BsAbs with high molecular weight aggregation less than 10% are selected for further characterization.

Example 5. Characterization of HIV- 1 RT BsAb antibodies

Binding affinity ofHIV-1 RT BsAb antibodies

[0575] The binding affinity of HIV- 1 RT BsAbs to recombinant HIV-1 RT/HLA-A*02:01 complex is measured, for example, by Surface Plasma Resonance (BiaCore). The binding parameters between the HIV-1 RT BsAb and the HIV-1 RT/HLA-A*02:01 complex are measured, for example, using a His Capture Kit (GE Healthcare, Cat# 28995056) on a Biacore XI 00 (GE Healthcare) according to the manufacturer's protocol for multi-cycle kinetics measurement. All of the proteins used in the assay are diluted using HBS-E buffer. For example, 1 μg/mL of the HIV-1 RT BsAb is immobilized onto a Sensor Chip pre-functionalized with the anti-histidine antibody by flowing the solution through flow cell 2 at 2 μί/ητίη for 2

minutes. Binding towards the HIV-1 RT/A*02:01 complex is analyzed at, for example, 0.19, 0.38, 7.5, 15, and 30 μg/mL, each run consisting of a 3 minute association and 3 minute dissociation at 30 μί/ητίη. At the end of the cycles, the surface is regenerated using the regeneration buffer from the His Capture kit. The data are analyzed using 1: 1 binding site model with the BiaCore X-100 evaluation software. The binding parameters (association on rate constant k_a, dissociation constant k_d, and equilibrium dissociation constant Ε¾ are then calculated.

T-cell killing assay with peptide-pulsed T2 cells

[0576] Tumor cytotoxicity was assayed by LDH Cytotoxicity Assay (Pro mega). Human T cells purchased from AllCells were activated and expanded with CD3/CD28 Dynabeads (Invitrogen) according to manufacturer's protocol. Activated T cells (ATC) were cultured and maintained in RPMI1640 medium with 10% FBS plus 30 U/ml IL-2, and used at day 7-14. The T cells were > 99% CD3⁺ by FACS analysis. Activated αβ T cells (effector cells) and target peptide- loaded T2 cells were co-cultured at a 5: 1 ratio with 1 μg/ml or 0.2 μg/ml of BsAbs for 16 hours. Peptide- loaded T2 cells were prepared by incubating T2 cells overnight with 50 μg/ml of either the target HIV-1 RT 181 M184V (YQYVDDLYV, SEQ ID NO: 6) or control peptide mixture P20 (SEQ ID NOs: 255-274). Cytotoxicities were then determined by measuring LDH activity in culture supernatants. As shown in FIG. 2, BsAbs 1, 4, 9, 10, 13, 14, 17, 27, 30, and 34 killed the HIV-1 RT peptide- loaded T2 cells in a selective manner.

T-cell killing assay with cell lines

[0577] Target cell cytotoxicity was assayed by LDH Cytotoxicity Assay (Pro mega). Human T cells were purchased from AllCells and activated and expanded with CD3/CD28 Dynabeads (Invitrogen) according to manufacturer's protocol. Activated T cells (ATC) were cultured and maintained in RPMI1640 medium with 10% FBS plus 30 U/ml IL-2, and used at day 7-14. Activated T cells (effector cells) and target cells were co-cultured at a 5: 1 ratio with different concentrations of BsAbs (including for example 0.2, 0.04, 0.008, and 0.0016 μg/ml BsAbs) for 16 hours. Cytotoxicities were then determined by measuring LDH activities in culture supernatants. [0578] Target cells tested included parental SK-Hepl cells and SK-Hepl cells transfected with a minigene to stably express HIV-1 RT WT (SEQ ID NO: 1) (SK-Hepl HIV-1 RT WT), HIV-1 RT M184V (SEQ ID NO: 3) (SK-Hepl HIV-1 RT M184V), or HIV-1 RT M184I (SEQ ID NO: 4) (SK-Hepl HIV-1 RT Ml 841). As shown in FIG. 3, BsAbs 1, 9, 10, 13, 14, 27, and 34 were able to specifically direct killing of SK-Hepl WT, M184V, and M184I cells, but not the parental SK-Hepl cells. BsAb clone 4 was able to specifically direct killing of SK-Hepl M184V and M184I cells, but not SK-Hepl WT cells. BsAbs 17 and 30 showed evidence of non-specific killing.

[0579] Additional target cells that may be tested include T cells, such as CD4⁺ T cells, transduced to express HIV-1 RT WT (SEQ ID NO: 1), HIV-1 RT M184V (SEQ ID NO: 3), or HIV-1 RT M184I (SEQ ID NO: 4).

Cross-reactivity of HIV-1 RT BsAb antibodies against multiple HLA-A02 alleles

[0580] Human MHCI molecules consist of 6 class isoforms, HLA-A, -B, -C, -E, -F and G. The HLA-A, -B and-C heavy chain genes are highly polymorphic. For each isoform, the HLA genes are further grouped according to the similarity of heavy chain sequences. For example, HLA-A is divided into different alleles such as HLA- AO 1, -A02, -A03, etc. For the HLA-A02 allele, there are multiple subtypes, such as HLA-A*02:01, A*02:02, etc. Between the different subtypes of HLA-A02 group, the sequence differences are limited to only several amino acids. So in many cases, peptides that bind to HLA-A*02:01 molecule can also form complexes with multiple subtypes of the HLA-A02 allele. As shown in Table 12

(http://www.allelefrequencies.net/), although HLA-A*02:01 is the dominant HLA-A02 subtype among Caucasian populations, in Asia, A*02:05, A*02:06, A*02:07 and A*02: l l are also common HLA-A02 subtypes. The ability of HIV-1 RT antibodies to recognize not only HIV-1 RT peptide in the context of HLA-A*02:01, but also other subtypes of HLA-A02, will greatly broaden the patient population that might be able to benefit from HIV-1 RT antibody drug treatment. To determine cross-reactivity, HIV-1 RT/MHC class I complexes with other subtypes of the HLA-A02 allele are generated and the binding affinity of the HIV-1 RT/HLA-A*02:01- specific antibodies for these other complexes is tested. Binding affinity is determined, for example, using a ForteBio Octet QK. Briefly, 5 μg/mL biotinylated HIV-1 RT peptide/HLA- A02 MHC complex having varying subtypes is loaded onto a streptavidin biosensor. After washing off excess antigen, BsAbs are tested at, for example, 10 μg/mL for association and dissociation kinetics. Binding parameters are calculated using a 1: 1 binding site, partial fit model. Table 12

australia china europe india north sub- taiwan us africa saharan

africa

A*02:01 97.8% 39.5% 94.0% 53.9% 73.3% 56.3% 35.1% 79.4%

A*02:02 0.0% 0.1% 0.3% 0.9% 9.7% 24.1% 0.0% 3.6%

A*02:03 0.0% 15.3% 0.2% 4.9% 0.0% 0.4% 19.3% 2.2%

A*02:04 0.0% 0.1% 0.0% 0.3% 2.6% 0.4% 0.0% 0.2%

A*02:05 1.1% 0.9% 3.2% 5.8% 13.8% 15.9% 0.1% 4.5%

A*02:06 0.0% 16.0% 0.9% 10.6% 0.0% 0.7% 12.8% 5.5%

A*02:07 1.1% 26.1% 0.4% 0.4% 0.0% 0.0% 32.7% 2.4%

A*02:08 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0%

A*02:09 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0%

A*02:10 0.0% 1.1% 0.0% 0.0% 0.0% 0.2% 0.0% 0.1%

A*02:ll 0.0% 0.1% 0.1% 22.3% 0.0% 1.5% 0.0% 1.7% other A02 subtypes 0.0% 0.7% 0.8% 0.9% 0.5% 0.5% 0.0% 0.6%

(A*02:12 - A*02:93)

100.0% 100.0% 100.0% 100.0% 100.0% 100.0% 100.0% 100.0%

Stability of HIV- 1 RT BsAb antibodies

[0581] Anti-RTMC BsAb clones #10, #14, and #27 were stored in a formulation buffer (30 mM Citric Acid, 200 mM Lysine, 0.05% polysorbate 80, pH 7) at 4°C for two years, and a target cell killing assay as described above was performed (using SK-Hepl target cells expressing HIV-1 RT 181 WT minigene). The percent specific lysis of clone #10 was found to be ~ 48% (64% of the 75% level determined two years prior as shown in FIG. 3), the percent specific lysis of clone #14 was found to be ~ 52% (69% of the 75% level determined two years prior as shown in Fig. 3), and the percent specific lysis of clone #27 was found to be ~ 4% (5% of the 83% level determined two years prior).

Example 6A. Generation of HIV-1 RT/HLA-A*02:01 specific chimeric antigen receptor (CAR) constructs

[0582] Chimeric antigen receptor therapy (CAR-T therapy) is a relatively new form of targeted immunotherapy. It merges the exquisite targeting specificity of monoclonal antibodies with the potent cytotoxicity and long-term persistence provided by cytotoxic T cells. This technology enables T cells to acquire long-term novel antigenic specificity independent of the endogenous TCR. Clinical trials have shown clinically significant antitumor activity of CAR-T therapy in neuroblastoma (Louis C.U. et al, Blood 118(23):6050-6056), B-ALL (Maude S.L. et al, N. Engl. J. Med. 371(16): 1507-1517, 2014), CLL (Brentjens R.J. et al, Blood 118(18):4817-4828, 2011), and B cell lymphoma (Kochenderfer J.N. et al, Blood. 116(20):4099-4102, 2010). In one study, a 90% complete remission rate in 30 patients with B-ALL treated with CD19-CAR T therapy was reported (Maude S.L. et al., supra).

[0583] To further explore the potency of the HIV-1 RT/HLA-A*02:01 specific antibodies, anti- RTMC scFv-containing CARs are constructed and transduced into T cells. For example, HIV-1 RT/HLA-A*02:01 specific CARs are constructed using a lentiviral CAR expression vector. Anti-HIV-1 RT/HLA-A*02:01 scFvs are grafted onto a second generation CAR (Mackall C.L. et al., Nat. Rev. Clin. Oncol. l l(12):693-703, 2014) with a co-stimulatory signaling domain (such as from CD28 or 4- IBB) and a signaling domain from TCRζ engineered in cis to provide intracellular T cell stimulation signals and to activate T cells. For example, the anti- HIV-1 RT/HLA-A*02:01 scFvs are grafted onto a CAR polypeptide having the amino acid sequence of SEQ ID NO: 279 or 280.

Example 6B. Generation of HIV-1 RT/HLA-A*02:01 specific chimeric antigen receptor (CAR) constructs

[0584] HIV-1 RT/HLA-A*02:01 specific CARs were generated using scFv sequences of the HIV-1 RT/HLA-A*02:01-specific phage clones 10 and 14 to generate anti-HIV CAR 10 (SEQ ID NO: 254) and anti-HIV CAR 14 (SEQ ID NO: 275). The anti-HIV- 1 RT/HLA-A*02:01 CARs comprise the scFv sequence of an HIV-1 RT/HLA-A*02:01-specific phage clone (from N-terminus to C-terminus: light chain variable region (LCVR), scFv linker (SEQ ID NO: 276), heavy chain variable region (HCVR)) fused at its C-terminal end to a CAR polypeptide comprising a CD3ζ intracellular signaling sequence and a CD28 intracellular signaling sequence (SEQ ID NO: 279). The phage clone heavy and light chain variable regions used for generating the CARs are listed in Table 13.

Table 13

Example 7. Generation of HIV-1 RT/HLA-A*02:01 specific chimeric antibody/TCR receptor-presenting T cells (abTCR T cells)

[0585] Chimeric antibody/T cell receptors (abTCRs) are regulated by the naturally occurring machinery that controls TCR activation, and can thus avoid being constitutively activated and negative outcomes associated with such activation. abTCR T cells are expected to signal and respond to endogenous T-cell regulatory mechanisms and to demonstrate increased persistance in vivo.

[0586] To further explore the potency of the HIV-1 RT/HLA-A*02:01 specific antibodies, anti- RTMC abTCRs are constructed and transduced into T cells. For example, HIV-1 RT/HLA- A*02:01 specific abTCRs are constructed using one or more lentiviral abTCR expression vectors. Anti-HIV-1 RT/HLA-A*02:01 Fab-like antigen-binding modules are grafted onto a T cell receptor transmembrane module comprising T cell receptor subunit transmembrane domains to provide intracellular T cell stimulation signals through association with endogenous TCR- associated signaling molecules (such as CD35s, CD3y8, and ζζ) and to activate T cells. For example, the anti- HIV-1 RT/HLA-A*02:01 Fab-like antigen-binding modules are grafted onto abTCR polypeptides comprising the amino acid sequences of SEQ ID NOs: 301 and 302.

Example 8A. Characterization of anti-RTMC chimeric receptor T cells

In vitro Cytotoxicity study of anti-RTMC chimeric receptor T cells

[0587] Lentiviruses encoding HIV-1 RT/HLA-A*02:01 specific chimeric antigen receptors (CARs) or chimeric antibody/TCRs (abTCRs) are produced, for example, by transfection of 293T cells with anti-RTMC CAR vectors or anti-RTMC abTCR vectors, respectively. Human T- cells are used for transduction after 2-day stimulation with CD3/CD28 beads (Dynabeads®, Invitrogen) in the presence of interleukin-2 at 30 U/ml. Concentrated lentiviruses are applied to T-cells in Retronectin (Takara) coated 6-well plates for 72 hours. Transduction efficiency is assessed by FACS using biotinylated HIV-1 RT tetramer and PE-conjugated streptavidin.

Repeat FACS analyses are done at 72 hours and every 3-4 days thereafter.

[0588] Functional assessment of the transduced T cells (anti-RTMC CAR T cells or anti-RTMC abTCR T cells) is performed using LDH Cytotoxicity Assay. Effector-to-target ratios used include, for example, 5: 1 and 10: 1. The target cell lines may include, for example, SK-HEP-1 (ATCC HTB-52; HLA-A2⁺, HIV-1^"), T cell leukemia cell line Jurkat (ATCC TIB- 152; HLA- A2^", HIV-1^"), leukemia cell line K562 (ATCC CCL-243; HLA-A2^", HIV-1^"), and primary T cells, such as CD4⁺ T cells. SK-HEP-1, Jukat, K562, and primary T cells are transduced with an HIV-1 RT-expressing minigene cassette encoding WT HIV-1 RT (SEQ ID NO: 1), M184V HIV-1 RT (SEQ ID NO: 3), or M184I HIV-1 RT (SEQ ID NO: 4), which results in a high level of cell surface expression of HIV-1 RT peptide/HLA-A*02:01 complex. The specificity and efficiency of the anti-RTMC CAR-expressing T cells or anti-RTMC abTCR-expressing T cells to kill the target-positive cells is determined as described above using LDH assay.

Viral suppression study of anti-RTMC chimeric receptor T cells

[0589] Anti-RTMC chimeric receptor T cells (anti-RTMC CAR T cells or anti-RTMC abTCR T cells) are produced as described above. PBMCs are treated with 1 mg/ml of PHA and lOng/ml of IL-2 in RPMI-1640 + 10% FBS for 2 days. CD4⁺ T cells are isolated by negative selection using magnetic beads and infected with multiple strains of HIV (for example NL4-3, Bal, and SF162). After 48 hours, cells are washed with fresh media. HIV-infected CD4⁺ T-cells are mixed with anti-RTMC CAR-transduced or anti-RTMC abTCR-transduced CD8⁺ T cells at exemplary effector-to-target ratios of 5: 1 or 1: 1. Every 2 days (for 8 days) intracellular p24 (Gag) levels are measured by flow cytometry. Briefly, HIV-infected CD4⁺ T cells are stained with antibodies to CD4, CD8, and p24 using, for example, the FIX & PERM® Cell Fixation & Cell Permeabilization Kit. For examples of viral suppression studies see Varela-Rohena, A. et al. (2008) Nature medicine 14(12): 1390-1395.

Proliferation of anti-RTMC chimeric receptor-transduced CD8⁺ T cells in response to HIV- infected CD4⁺ T cells

[0590] The purpose of this experiment is to measure the proliferative capacity of CD8⁺ T cells transducted with either an anti-RTMC CAR or an anti-RTMC abTCR in response to exposure to HIV-infected CD4⁺ T cells.

[0591] HIV-infected CD4⁺ T-cells are produced as described above. Target cells are irradiated, for example, with high-dose gamma-radiation (e.g., 10,000 rad). Anti-RTMC chimeric receptor T cells (anti-RTMC CAR T cells or anti-RTMC abTCR T cells) are labeled with Cell Trace Violet (Thermo Fisher) according to the manufacturer's instructions. Target cells and anti- RTMC chimeric receptor T cells are mixed, for example, at 1: 1 ratio. The media is changed after 3 days and Cell Trace Violet fluorescence is measured after 7 days. For examples of proliferation studies see Ali, A. et al. (2016) Journal of virology JVI-00805.

[0592] Cytokine release by activated anti-RTMC chimeric receptor-transduced CD8⁺ T cells

[0593] The purpose of this assay is to measure cytokine release by activated CD8⁺ anti-RTMC chimeric receptor T cells (anti-RTMC CAR T cells or anti-RTMC abTCR T cells) in response to HIV-infected CD4⁺ T cells. For examples of analysis of cytokine production by transduced CD8⁺ T cells see Varela-Rohena, A. et al. (2008) Nature medicine 14(12): 1390-1395.

[0594] HIV-infected CD4⁺ T-cells are produced as described above. Target cells and anti- RTMC chimeric receptor T cells are mixed, for example, at 1: 1 or 1:8 exemplary E:T ratios. Cytokines released by CD8⁺ chimeric receptor T cells are analyzed, for example, using ELISA Kits for Granzyme B (Neobio science), IL-2 (MultiSciences, Lianke Biotech), and IFN-γ. See for example Liu, B. et al. (2016) Journal of Virology JVI-00852.

Example 8B. Characterization of anti-RTMC CAR T cells

In vitro Cytotoxicity study of anti-RTMC chimeric receptor T cells

[0595] Lentiviruses encoding HIV-1 RT/HLA-A*02:01 specific CARs were produced by transfection of 293T cells with anti-RTMC CAR vectors encoding anti-HIV CAR 10 (SEQ ID NOs: 254) and anti-HIV CAR 14 (SEQ ID NO: 275). Human T-cells were used for transduction after 1-day stimulation with CD3/CD28 beads (Dynabeads®, Invitrogen) in the presence of interleukin-2 at 30 U/ml. Concentrated lentiviruses (MOI 3: 1) were applied to T-cells in

Retronectin (Takara) coated 6-well plates for 72 hours. Mock-transfected T cells were also prepared for use as a control. Transduction efficiency was assessed by FACS using biotinylated HIV-1 RT tetramer and PE-conjugated streptavidin (FIG. 4).

[0596] Functional assessment of the transduced T cells (anti-HIV CAR 10 T cells or anti-HIV CAR 14 T cells) and mock T cells grown in IL-2 at day 9 was performed using LDH

Cytotoxicity Assay. An effector-to-target ratio of 5: 1 with approximately 1 x 10⁵ target cells per well was used. The target cell lines included SK-HEP-1 control cells (ATCC HTB-52; HLA- A2⁺, HIV-Γ) and SK-HEP-l-MG, which was prepared by transducing parental SK-HEP-1 cells with an HIV-1 RT peptide-expressing minigene cassette encoding WT HIV-1 RT 181-189 (SEQ ID NO: 5), resulting in a high level of cell surface expression of HIV-1 RT 181-189/HLA- A*02:01 complex.

[0597] The specificity and efficiency of the anti-RTMC CAR-expressing T cells to kill the target-positive cells was determined as described above using LDH assay (FIG. 5). Both of the anti-RTMC CAR-expressing T cells killed the target-positive SK-HEP-l-MG cells in a specific and highly efficient way. Target-negative, wild-type SK-HEP1 cells, however, were poorly recognized by the same T cells. Theses results demonstrates that the antibody clones are highly specific to HIV RT-expressing cells and do not mediate non-specific killing of cells lines that do not express HIV RT.

Example 9. Generation and Characterization of the full-length IgGl HIV-1 RT antibodies

[0598] Full-length human IgGl of the selected phage clones are produced, for example, in

HEK293 and Chinese hamster ovary (CHO) cell lines, as described (Tomimatsu K. et al., Biosci. Biotechnol. Biochem. 73(7): 1465- 1469, 2009). In brief, antibody variable regions are subcloned into mammalian expression vectors, with matching human lambda or kappa light chain constant region and human IgGl constant region sequences. Applying the same cloning strategy, chimeric anti-HIV-1 RT peptide/MHC full-length antibodies with mouse IgGl heavy chain and light chain constant regions are generated. Molecular weight of the purified full length IgG antibodies is measured under both reducing and non-reducing conditions by electrophoresis. SDS-PAGE of purified HIV-1 RT mouse chimeric IgGl antibodies is performed to determine protein purity. In brief, 2μg of the protein is mixed with 2.5μί of NuPAGE LDS Sample Buffer (Life Technologies, NP0008) and brought up to ΙΟμί with deionized water. The sample is heated at 70°C for 10 minutes, and then loaded onto the gel. Gel electrophoresis is performed at 180V for 1 hour.

[0599] Anti-HIV-1 RT peptide/MHC chimeric IgGl antibody is tested for binding towards HIV- 1 RT presenting cells (such as SK-Hepl, Jurkat, K562, and primary CD4⁺ cells transduced with a minigene cassete as described above) by flow cytometry. An HIV-1 RT minigene cassette (expressing WT, M184V, or M184I HIV-1 RT) is transfected into cells to generate the HIV-1 RT-presenting target cells. 10 μg/mL of antibody is added to cells on ice for 1 hour. After washing, R-PE conjugated anti-mouse IgG(H+L) (Vector Labs#EI-2007) is added to detect antibody binding. Binding affinity of the mouse chimeric IgGl anti-HIV-1 RT peptide/MHC antibodies is determined by ForteBio Octet QK. 5 μg/mL biotinylated HIV-1 RT peptide/HLA- A*02:01 complex is loaded onto a streptavidin biosensor. After washing off excess antigen, mouse chimeric full-length antibodies are tested at 10 μg/mL for association and dissociation kinetics. Binding parameters are calculated using a 1: 1 binding site, partial fit model.

[0600] HIV-1 RT-specific and negative control mouse chimeric IgGl are tested for binding towards HIV-1 RT/HLA-A*02:01, HIV-1 RT recombinant protein and free HIV-1 RT 181 peptide in an ELISA assay. Antibodies are tested, for example, at 3x serial dilution, starting from 100 ng/mL for a total of 8 concentrations. Biotinlyated HIV-1 RT/A*02:01 MHC is coated onto streptavidin plates at 2 μg/mL, HIV-1 RT protein is coated at 2 μg/mL and HIV-1 RT peptide is coated at 40 ng/mL. The ability of full-length anti-HIV-1 RT/HLA-A*02:01 antibodies to recognize the HIV-1 RT peptide only in the context of HLA-A02, and not bind recombinant HIV-1 RT protein or free HIV-1 RT peptide is determined.

Example 10. In vivo efficacy studies

Anti-RTMC chimeric receptor T cell treatment in mice [0601] Anti-RTMC chimeric receptor T cells (such as anti-RTMC CAR T cells or anti-RTMC abTCR cells) are evaluated in a mouse model of HIV infection. For examples of such mouse models see Akkina, R. (2013) Virology 435(1): 14-28. For example, NOD SCID gamma (NSG) mice are engrafted with human peripheral blood mononuclear cells (PBMCs), human

hematopoietic stem cells (HSCs), thymus cells, liver cells, and/or bone marrow cells from an HLA-A2⁺ source to create humanized NSG mice. The mice are then infected with HIV, such as by rectal or vaginal inoculation with HIV-1. Mice are divided into at least 3 groups that receive one of the following: (i) no treatment; (ii) mock treatment; or (iii) treatment with an anti-RTMC construct (e.g., BsAb) or anti-RTMC chimeric receptor T cells (anti-RTMC CAR T cells or anti- RTMC abTCR T cells). The animals in each group are monitored for HIV-associated disease pathologies, CD4⁺ T cell levels, HIV viral load, body weight, and general health condition (eating, walking, daily activities).

Example 11. Affinity Maturation of anti-HIV-1 RT antibody agents

[0602] This example describes the affinity maturation of anti-HIV-1 RT antibody agents. In particular, this example specifically describes the generation of a series of antibody variants by incorporation of random mutations into a representative anti-HIV-1 RT antibody followed by screening and characterization of the antibody variants.

[0603] Generation of variant phage libraries

[0604] DNA encoding an anti-HIV-1 RT peptide/MHC scFv is subjected to random mutagenesis using GeneMorph II Random Mutagenesis kit (Agilent Technologies) according to the manufacturer's specifications. After mutagenesis, DNA sequences are cloned into an scFv- expressing phagemid vector to build a variant human antibody phage library which contains, for example, about 5x10 unique phage clones. On average, variant clones have two nucleotide mutations compared with the parental anti-HIV-1 RT peptide/MHC clone, ranging from 1 to 4 nucleotide mutations, per scFv sequence.

Cell panning

[0605] The human phage scFv library with mutants is used to pan against HIV-1 RT 181 peptide/HLA-A*02:01 complex as described in Example 2. In particular, cell panning is used.

For example, human scFv phage library is first mixed with T2 cells loaded with 50 ug/ml of a pool of 20 different endogenous peptides (P20, SEQ ID NOs: 255-274) as negative control panning. The negative control-depleted human scFv phage library is then mixed with T2 cells loaded with HIV-1 RT 181 peptide (1.5 ug/ml first round, 0.8 ug/ml second round, 0.4 ug/ml third round) for positive selection. To load peptide, T2 cells are pulsed with peptides in serum- free RPMI1640 medium in the presence of 20 μg/ml β2Μ overnight. After extended washing with PBS, peptide- loaded T2 cells with bound scFv antibody phage are spun down. The bound clones are then eluted and used to infect E.coli XLl-Blue cells. The phage clones expressed in bacteria are then purified. The panning is performed for 3 rounds to enrich for scFv phage clones that bind HIV-1 RT 181 peptide/HLA-A*02:01 specifically.

[0606] Streptavidin ELISA plates are coated with biotinylated HIV-1 RT 181 peptide/HLA- A*02:01 complex monomer or biotinylated P20 control peptides/HLA-A*02:01 monomer. Individual phage clones from enriched phage display panning pools against HIV-1 RT 181 peptide/HLA-A*02:01 complex are incubated in the coated plates. Binding of the phage clones is detected by HRP-conjugated anti-M13 antibodies and developed using HRP substrate. The absorbance is read at 450nm. Positive clones are identified through ELISA screening of phage clones enriched from phage panning. Unique clones are identified by DNA sequencing of the ELISA-positive phage clones. Specific and unique clones are further tested for their binding to HLA-A*02:01/peptide complexes on the live cell surface by flow cytometry (FACS analysis) using HIV-1 RT 181 peptide- loaded live T2 cells. Controls include T2 cells without peptide loading (cells only) and R-PE conjugated horse anti-mouse IgG control (secondary antibody only). Briefly, T2 cells loaded with HIV-1 RT 181 peptide or P20 peptide pool are first stained with purified scFv phage clones, followed by a second staining with mouse anti-M13 mAb, and a third staining with R-PE conjugated horse anti-mouse IgG from Vector Labs. Each staining step is performed for 30-60 minutes on ice and the cells are washed twice between staining steps. Specific binding to HIV-1 RT 181-loaded T2 cells and not T2 cells loaded with P20 peptide pool in the context of HLA-A*02:01, or T2 cells without peptide loaded is determined.

Example 12. Characterization of bi-specific antibody molecules based on anti-HIV-1 RT affinity maturation variants

Generation of bispecific antibodies

[0607] Bispecific antibodies (BsAbs) are generated using scFv sequences of the affinity matured HIV-1 RT/HLA-A*02:01-specific phage clones isolated in Example 10 using the method described in Example 4. The resulting single-chain bispecific antibodies comprise the scFv sequence of an HIV-1 RT/HLA-A*02:01-specific phage clone at the N-terminal end and an anti- human CD3s mouse monoclonal scFv at the C-terminal end.

Determination of binding affinities of bispecific antibodies to HIV-1 RT/HLA-A*02:01 [0608] The binding affinity of HIV- 1 RT BsAb antibodies (derived from affinity matured clones) to recombinant HIV-1 RT/HLA-A*02:01 complex is measured by Surface Plasmon Resonance (BiaCore). The binding parameters between the HIV-1 RT BsAbs and HIV-1 RT/HLA-A*02:01 complex are measured using a Biotin CAPture Kit (GE Healthcare, Cat# 28920233) on a Biacore X100 (GE Healthcare) according to the manufacturer's protocol for multi-cycle kinetics measurement. All of the proteins used in the assay are diluted using HBS- EP running buffer. 5 μg/mL of biotinylated HIV-1 RT 181/HLA-A*02:01/p2M complex is immobilized onto a Sensor Chip CAP pre-functionalized with streptavidin (-3,800 RU of streptavidin captured) by flowing the solution through the flow cell at 5 μΕ/ητίη for 75 seconds (-120 RU of MHC complex was captured per cycle). Binding towards the HIV-1 RT BsAbs is analyzed at 150 nM, 75 nM, 37.5 nM, 18.8 nM, and 9.4 nM, each run consisting of a 2 minute association and 10 minute dissociation at 30 μΕ/ητίη. At the end of cycle, the surface is regenerated using the regeneration buffer from the Biotin CAPture kit. The data is analyzed using 1: 1 binding site model with the BiaCore X-100 evaluation software. The binding parameters (association on rate constant k_a, dissociation constant k_d, and equilibrium dissociation constant K_d) are then calculated.

Cross-reactivity and binding affinities of HIV-1 RT bispecific antibodies against multiple HLA- A02 alleles

[0609] As described above in Example 5, the different subtypes of the HLA-A02 group are quite conserved, and cross-reactivity against multiple HLA-A02 subtypes is highly desired. Therefore, experiments are performed to determine whether the bi- specific antibodies (BsAb) generated from the parental clone and the affinity maturation variants cross-react with non-HLA-A*02:01 subtypes of the HLA-A02 group. Specifically, HIV-1 RT 181/MHC class I complexes with various subtypes of the HLA-A02 allele are generated, and their binding affinities to the HIV-1 RT 181/HLA-A*02:01-specific antibodies are determined using the Octet® QKe System by Pall ForteBio LLC (Menlo Park, CA), which utilizes the Biolayer Interferometry (BLI) technology. The BsAbs tested include the parental clone and affinity maturation variant clones. Five μg/mL biotinylated HIV-1 RT 181 peptide/HLA-A02 complex having varying subtypes of HLA-A02 is loaded onto a streptavidin biosensor. After washing off excess antigen (the HIV-1 RT

peptide/HLA-A02 complex), BsAbs are tested at 10 μg/mL for association and dissociation kinetics. Binding parameters are calculated using a 1: 1 binding site, partial fit model.

Peptide binding specificity assay [0610] In order to confirm the specificity of the peptide recognized by the affinity maturation variant antibodies, a FACS analysis is performed with T2 cells loaded with HIV-1 RT 181 peptide, the P20 peptide pool, or no peptide.

Epitope mapping by alanine walking

[0611] To investigate with precision the sensitive residues of the HIV-1 RT 181 peptide for recognition by BsAb affinity maturation variants, alanine walking experiments as described above are performed.

Sequence Listing

GDAYFSVPLDKDFRKYTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKI

LEPFRKQNPDIVIYQYVDDLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQ

KEPPFLWMGYELHPDKWTVQPIVLPEKDSWTVNDIQKLVGKLNWASQIYAGIK

VRQLCKLLRGTKALTEVVPLTEEAELELAENREILKEPVHGVYYDPSKDLIAEIQKQ

GQGQWTYQIYQEPFKNLKTGKYARMKGAHTNDVKQLTEAVQKIATESIVIWGK

TPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPPLVKLWYQLEKEPIIGAE

TFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQAIHLALQDSGLE

VNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAHKGIGGNEQV

DKLVSAGIRKVL

PISPIETVPVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYN HIV-1 RTM184I

TPVFAIKKKDSTKWRKLVDFRELNKRTQDFWEVQLGIPHPAGLKQKKSVTVLDV amino acid

GDAYFSVPLDKDFRKYTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKI

LEPFRKQNPDIVIYQYIDDLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQK

EPPFLWMGYELHPDKWTVQPIVLPEKDSWTVNDIQKLVGKLNWASQIYAGIKV

RQLCKLLRGTKALTEVVPLTEEAELELAENREILKEPVHGVYYDPSKDLIAEIQKQG

QGQWTYQIYQEPFKNLKTGKYARMKGAHTNDVKQLTEAVQKIATESIVIWGKT

PKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPPLVKLWYQLEKEPIIGAET

FYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQAIHLALQDSGLEV

NIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAHKGIGGNEQVD

KLVSAGIRKVL

YQYMDDLYV HIV-1 RT 181-189

YQYVDDLYV HIV-1 RT 181-189

M184V

YQYIDDLYV HIV-1 RT 181-189

M184I

CQYMDDLYV HIV-1 RT 181-189

Y181C

CQYVDDLYV HIV-1 RT 181-189

Y181C, M184V

AQYMDDLYV HIV-1 RT 181-189 Al

YAYMDDLYV HIV-1 RT 181-189 A2

YQAMDDLYV HIV-1 RT 181-189 A3

YQYADDLYV HIV-1 RT 181-189 A4

YQYMADLYV HIV-1 RT 181-189 A5

YQYMDALYV HIV-1 RT 181-189 A6

YQYMDDAYV HIV-1 RT 181-189 A7

YQYMDDLAV HIV-1 RT 181-189 A8

YQYMDDLYA HIV-1 RT 181-189 A9

EVQLVESGGGVVRPGGSLRLSCAASGFTFGDYGMSWVRQAPGKGLEWVSGIN HCVR 1

WNGGSTGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCARDPIYSGSY KAFDYWGQGTLVTVSS

EVQLVQSGGVLVQPGGSLRLSCAASGFTFSNYAMTWVRQAPGEGLEWVSGISR HCVR3

TGATISYADSVKGRFTISRDNSKNTLHLQMNSLRDEDTAIYYCVKDLFGADSGYD

GVAYYFGDWGQGTLVTVSS

EVQLVQSGAEVKKPGASVKISCKASGYYFSAYYIHWVRQAPGHGLEWVGFINPS HCVR 4

GGSTSYAQKFRGRITMTGDTSTSTVDMELSNLRSEDTAVYYCARGSDYYPFNH

WGDLWGQGTLVTVSS

EVQLVESGGGLVKPGGSLRLSCAGSGFTFSDYYMNWIRQAPGKGLEWISYITGK HCVR 5

SSYTNYADSVKGRFTISRDNAKNILYLQMNNLRAEDTAVYYCARYPSYVEGDYW GQGTLVTVSS

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWIS HCVR 6 AYNGNTNYAQKLQGRVTMTTDTSTSTAYM ELRSLRSDDTAVYYCARDSYYFDK WGQGTLVTVSS

QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGVNWVRQAPGQGLEWMGWI HCVR 7

SPYN DYTNYAPNVQGRVTMTTDTSTNTAYLEVRSLRSDDTAVYYCARSFYDSW

GQGTLVTVSS

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWI N HCVR 8

TYNGNTNYPQKLQGRVTMTRDTSTSTAYM ELSSLRSDDTAVYYCARYSSTGRW

QDWWGQGTLVTVSS

EVQLVQSGAEVKKPGESLRISCKGSGYSFTSYWISWVRQM PGKGLEWMGRI DP HCVR 9 SDSYTNYSPSFQGHVTISADKSISTAYLQWSSLKASDTAMYYCARIGGMVKDTW GQGTLVTVSS

EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRI I PI HCVR 10

LDI PNYAQKFQGRVTITADNSTSTAYM ELSSLRSEDTAVYYCARGYGWQSYDG

WGQGTLVTVSS

QMQLVQSGSELKKAGASVKVSCKASGYTFTDYSI NWVRQAPGQGLEWMGW HCVR 12 M NTNTGKPTYAQGFTGRFVFSLDTSVSTAYLQISSLKPEDTAVYYCARSSGDYW GQGTLVTVSS

EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDI NWVRQATGQGLEWMGWM HCVR 13 N PNSGNTGYAQKFQGRVTMTRNTSISTAYM ELSSLRPEDTAVYYCARSDFDSW GQGTLVTVSS

QVQLVQSGAEVRKPGASVKVSCKASGYI FNNYYI HWVRQAPGQGLEWMGWI HCVR 14 NTYTG N PTYAQG FTG R FVFS LDTSVSTAYLQI SS LKAE DTAVYYC ARGSYS WSYYS QKDYWGQGTLVTVSS

EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWIS HCVR 15 AYNGNTNYAQKLQGRVTMTTDTSTSTAYM ELRSLRSDDTAVYYCARDAYVYDS WGQGTPVTVSS

QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWI RQAPGKGLEWVSYISSS HCVR 16 GSTIYYADSVKGRFTISRDNAKNSLYLQM NSLRAEDTAVYYCARQGWYMYLGW DYWGQGTLVTVSS

EVQLVETGGGLVQPGGSLRLSCAASGLTFSMYAM NWVRQPPGKGLEWVAGIS HCVR 17 SSGGSTYYADSVKGRFTISRDNSKNTLYLEM NSLRAEDTAVYYCARGQHGSYYSY SDYWGQGTLVTVSS

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDI NWVRQATGQGLEWMGWM HCVR 18 N PNSGNTGYAQKFQGRVTMTRNTSISTAYM ELSSLRSEDTAVYYCARMSH RVG YMGAGFDPWGQGTLVTVSS

EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWM HCVR 19 N PHSGNTGYAQKFQGRVTMTRDTSTSTAYM ELSSLRSEDTAVYYCARSGFDIW GQGTLVTVSS

QVQLVQSGAEVKKPGASVKVSCKASGYPFTSYGISWVRQAPGQGLEWMGWIS HCVR 20 GYNGNTDYAQKLQGRVTMTTDTSTTTAYM ELRSLRSDDTAVYYCARWWHPW YGDH WGQGTLVTVSS

QMQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRI IP HCVR 21 I LGIANYAQKFQGRVTITADKSTSTAYM ELSSLRSEDTAVYYCARGSISYSIYFMSS

Dl WGQGTLVTVSS

QVQLQESGPGLVKPSETLSLTCTVSGGSISPYYWSWI RQPPGRGLEWIGYISDSG HCVR 22 TAKYN PSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGRSVMGYYYSDYW GQGTLVTVSS EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGFSWVRQATGQGLEWMGI I DP HCVR 24

SGGSTTYAQKFQGRVTMTRDTSTNTVYM ELSSLRSEDTAVYYCARQAVDQWG

QGTLVTVSS

EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGI I PI HCVR 26 FGTANYAQKFQGRVTITADESTSTAYM ELSSLRSEDTAVYYCARYRGSSLWYQYV DYWGQGTLVTVSS

EVQLVESGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGRI I PI L HCVR 27

GIANYAQKFQGRVTITADKSTSTAYM ELSSLRSEDTAVYYCARAYGRSYYDSWG

QGTLVTVSS

QVQLVESGAEVKKPGDSVKVSCKPSGYN FLNYGI NWVRQAPGQGLEWMGWIS HCVR 30 TYTGNTNYAQKLQGRVTFTTDTSTSTAYM EM RSLRSDDTAVYYCARSWGGYP WYSM DYWGQGTLVTVSS

EVQLVQSGAEVKKPGESLKISCQASGYAFSNYWIGWVRQMPGKGLEWIGM IYP HCVR 32 GDSDTRYSPSFQGHVTISADKSI NTAYLQWSSLKASDTAMYYCARGFWYRDGW GQGTLVTVSS

QVQLVQSGAEVKQPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWIS HCVR 34 AYNGNTNYAQKLQGRVTMTTDTSTSTAYM ELRSLRSDDTAVYYCARGVRSSQY DYWGQGTLVTVSS

QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYMSWI RQAPGKGLEWVSYISSS HCVR 37 GSTIYYADSVKGRFTISRDNAKNSLYLQM NSLRAEDTAVYYCARLFYYPH DDWW GQGTLVTVSS

QLQLQESGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWI RQSPSRGLEWLGRTY HCVR 39 YRSKWYN DYAVSVKS R ITI N P DTS KN QFS LQLN SVTP E DTAVYYC ARG LWSSYG F DNWGQGTLVTVSS

QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYYNSN LCVR 1 RPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGYVFGTGTKVTVL G

QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNS LCVR 3

N RPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGLYVFGTGTKVT

VLG

QSALTQPPSASGSPGQSVTISCTGTSSDVGGYNYVSWYQQH PGKAPKLM IYEVT LCVR 4 KRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYAGSKGVFGGGTKLTVL G

QSVLTQPPSVSGAPGQRVTISCTGSTSKLGPGYDVHWYQHVPGRAPKLLI H HNS LCVR 5 N RPSGVPDRFSGSKSGSSASLTITGLQAEDEADYYCQSYDTSLSGSVFGGGTKLTV LG

QSVLTQPPSASGTPGQRVTISCTGSSSDIGAGN DVHWH RQLPGTAPKLLIYGNS LCVR 6

N RPSGVPDRFSGSKSGTSASLVITGLQAEDEAVYYCQSYDGSLSGGSQVFGGGTK

LTVLG

DIQLTQSPSSVSASVGDRVTITCRASQGISRWLAWYQQKPGKAPKLLIYAASSLQ LCVR 7 SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANN FPLTFGGGTKVEI KR

QSVLTQPPSVSAAPGQKVTISCSGSSSN IGH NYVSWYQQLPGTAPKLLIYAHN ER LCVR 8 PSGI PDRFSGSKSGTSATLGITGLQTGDEADYFCGTWDDRLSGEVFGGGTKLTVL G

SYVLTQPPSVSAGPRQRVTISCSGSSSN I LN NAVNWYQQLPGMAPKLLIYYSDLL LCVR 9 SSGVSTRFSGSKSGTSASLAISGLQSEDEADYYCAVWDDSVKGYVFGTGTKVTVL G

QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKVLIYGNS LCVR 10 N RPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSHYVFGTGTK

252 GX39WDX40X41LSX42X43V LC-CDR3 consensus

X₃₉ = S or T, X₄₀ = any AA, X_4i = any AA, X₄₂ = A or G, X₄₃ = any AA 3

253 QSYDX44SLSG LC-CDR3 consensus X₄₄ = any AA 4

254 QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKVLIYGNS anti-HIVCAR 10

N PSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGSHYVFGTGTK

VTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGSSVKVSCKASG

GTFSSYAISWVRQAPGQGLEWMGRIIPILDIPNYAQKFQGRVTITADNSTSTAY

MELSSLRSEDTAVYYCARGYGWQSYDGWGQGTLVTVSSEQKLISEEDLAAAIEV

MYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVT

VAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFS

RSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEG

LYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALP

PR

255 YLLPAIVHI C3 control peptide

(A2E-7)

256 LLDVPTAAV A2E-1

257 TLWVDPYEV A2E-2

258 FLLDHLKRV A2E-3

259 LLLDVPTAAV A2E-4

260 VLFRGGPRGLLAV A2E-5

261 SLLPAIVEL A2E-6

262 FLLPTGAEA A2E-8

263 LLDPKLCYLL A2E-9

264 MLLSVPLLLG A2E-11

265 MVDGTLLLL A2E-17

266 SLPHFHHPET DMTN control

267 LLYDMVCGDIP PIM1 control

268 LLLDVPTAAVQ IFI30 control

269 LLLDVPTAAVQA IFI30 control

270 VLFRGGPRGLLAVA SSR1 control

271 YMAPEILMRS RPS6KB1 control

272 FIYNADLMNC CSF2RA control

273 KQYESVLMVSI IL7 control

274 KVNVDEVGGE Beta globin control

275 QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYSNNQR anti-HIVCAR 14

PSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNGYVFGTGTKVTVL

GSRGGGGSGGGGSGGGGSLEMAQVQLVQSGAEVRKPGASVKVSCKASGYIFN

NYYIHWVRQAPGQGLEWMGWINTYTGNPTYAQGFTGRFVFSLDTSVSTAYLQI

SSLKAEDTAVYYCARGSYSWSYYSQKDYWGQGTLVTVSSEQKLISEEDLAAAIEV

MYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVT

VAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFS

RSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEG

LYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALP

PR

276 SRGGGGSGGGGSGGGGSLEMA scFv linker

277 DVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYIN anti-CD3 scFv

PSRGYTNYADSVKGRFTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDY

WGQGTTVTVSSGEGTSTGSGGSGGSGGADDIVLTQSPATLSLSPGERATLSCRA SQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGSGTDYSLTINSLEA

EDAATYYCQQWSSN PLTFGGGTKVEI K

278 TSGGGGS BiTE linker

279 AAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLAC Οϋ28Α:ϋ3ζ

YSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRK

NPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALH

MQALPPR

280 TGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLA 4-1ΒΒ/0ϋ3ζ

GTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGC

ELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRR

KNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDAL

HMQALPPR

281 PNIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRS TCRa constant

MDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTN domain

LNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS

282 EDLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHS TCR constant

GVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDE domain

WTQDRAKPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLV

SALVLMAMVKRKDF

283 SQPHTKPSVFVMKNGTNVACLVKEFYPKDIRINLVSSKKITEFDPAIVISPSGKYNA TCR6 constant

VKLGKYEDSNSVTCSVQHDNKTVHSTDFEVKTDSTDHVKPKETENTKQPSKSCH domain

KPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFL

284 DKQLDADVSPKPTIFLPSIAETKLQKAGTYLCLLEKFFPDVIKIHWQEKKSNTILGS TCRy constant

QEGNTMKTNDTYMKFSWLTVPEKSLDKEHRCIVRHENNKNGVDQEIIFPPIKTD domain

VITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCLLRRTAFCC

NGEKS

285 1 LLLKVAGFN LLMTLRLWSS TCRa

transmembrane domain

286 Tl LYEILLG KATLYAVLVSALVL TCR

transmembrane domain

287 MLFAKTVAVNFLLTAKLFFL TCR6

transmembrane domain

288 YYMYLLLLLKSVVYFAIITCCLL TCRy

transmembrane domain

289 ESSCDVKLVEKSFETDTNLNFQNLSVIGFR TCRa connecting peptide

290 ADCGFTSVSYQQGVLSA TCR connecting peptide

291 DHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLR TCR6 connecting peptide

292 MDPKDNCSKDANDTLLLQLTNTSA TCRy connecting peptide

293 IPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFR TCRa connecting peptide MD 294 GRADCGFTSVSYQQGVLSA TCR connecting peptide MD

295 EVKTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLR TCR6 connecting peptide MD

296 PIKTDVITMDPKDNCSKDANDTLLLQLTNTSA TCRy connecting peptide MD

297 MAMVKRKDF TCR intracellular domain

298 RRTAFCCNGEKS TCRy intracellular domain

299 M ETDTLLLWVLLLWVPGSTG Signal peptide

300 IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLL fragment of CD28 VTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

301 EVKTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKT T cell receptor VAVNFLLTAKLFFL domain, delta

302 PIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCLLRR T cell receptor TAFCCNGEKS domain, gamma

Claims

CLAIMS What is claimed is:

1. An isolated anti-RTMC construct comprising an antibody moiety that specifically binds to a complex comprising human immunodeficiency virus 1 (HIV-1) reverse transcriptase (RT) peptide and a major histocompatibility (MHC) class I protein ("RTMC").

2. The isolated anti-RTMC construct of claim 1, wherein the MHC class I protein is HLA- A*02:01.

3. The isolated anti-RTMC construct of claim 1 or 2, wherein the HIV-1 RT peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-18.

4. The isolated anti-RTMC construct of claim 3, wherein the HIV-1 RT peptide has an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-9.

5. The isolated anti-RTMC construct of claim 1 or 2, wherein the antibody moiety specifically binds to one or more of:

i) a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of SEQ ID NO: 5 and the MHC class I protein;

ii) a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of SEQ ID NO: 6 and the MHC class I protein;

iii) a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of SEQ ID NO: 7 and the MHC class I protein;

iv) a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of SEQ ID NO: 8 and the MHC class I protein; and

v) a complex comprising an HIV-1 RT peptide comprising the amino acid sequence of SEQ ID NO: 9 and the MHC class I protein.

6. The isolated anti-RTMC construct of any one of claims 1-5, wherein the antibody moiety is a full-length antibody, a Fab, a Fab', a (Fab')2, an Fv, or a single chain Fv (scFv).

7. The isolated anti-RTMC construct of any one of claims 1-6, wherein the isolated anti- RTMC construct binds to the HIV-1 RT/MHC class I complex with a Kd from about 0.1 pM to about 500 nM.

8. The isolated anti-RTMC construct of any one of claims 1-7, wherein the antibody moiety comprises:

i) a heavy chain variable domain comprising a heavy chain complementarity determining region (HC-CDR) 1 comprising the amino acid sequence of SEQ ID NO: 240, or a variant thereof comprising up to about 3 amino acid substitutions, an HC-CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 241-244, or a variant thereof comprising up to about 3 amino acid substitutions, and an HC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 245-246, or a variant thereof comprising up to about 3 amino acid substitutions; and

ii) a light chain variable domain comprising a light chain complementarity determining region (LC-CDR) 1 comprising the amino acid sequence of any one of SEQ ID NOs: 247-249, or a variant thereof comprising up to about 3 amino acid substitutions, and an LC-CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 250-253, or a variant thereof comprising up to about 3 amino acid substitutions.

9. The isolated anti-RTMC construct of any one of claims 1-7, wherein the antibody moiety comprises:

10. The isolated anti-RTMC construct of claim 8 or 9, wherein the antibody moiety comprises a) a heavy chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 19-46, or a variant thereof having at least about 95% sequence identify to any one of SEQ ID NOs: 19-46; and b) a light chain variable domain comprising the amino acid sequence of any one of SEQ ID NOs: 47-74, or a variant thereof having at least about 95% sequence identity to any one of SEQ ID NOs: 47-74.

11. The isolated anti-RTMC construct of any one of claims 1- 10, wherein the isolated anti- RTMC construct is multispecific.

12. The isolated anti-RTMC construct of claim 11, wherein the isolated anti-RTMC construct is a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into-hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a heteromultimeric antibody, or a heteroconjugate antibody.

13. The isolated anti-RTMC construct of claim 11 or 12, wherein the isolated anti-RTMC construct further comprises a second antibody moiety that specifically binds to a second antigen.

14. The isolated anti-RTMC construct of claim 13, wherein wherein the second antigen is selected from the group consisting of CD3y, CD35, CD3s, ΟΌ3ζ, CD28, OX40, GITR, CD137, CD27, CD40L and HVEM.

15. The isolated anti-RTMC construct of claim 14, wherein the second antigen is CD3s, and wherein the isolated anti-RTMC construct is a tandem scFv comprising an N-terminal scFv specific for the HIV- 1 RT/MHC class I complex and a C-terminal scFv specific for CD3s.

16. The isolated anti-RTMC construct of any one of claims 1- 10, wherein the isolated anti- RTMC construct is a chimeric antigen receptor (CAR).

17. The isolated anti-RTMC construct of any one of claims 1- 10, wherin the isolated anti- RTMC construct is a chimeric antibody/T cell receptor (abTCR) comprising an extracellular domain comprising the antibody moiety and a T cell receptor (TCR) module (TCRM) comprising TCR transmembrane domains.

18. The isolated anti-RTMC construct of any one of claims 1- 10, wherein the isolated anti- RTMC construct is an immunoconjugate comprising the antibody moiety and an effector molecule, wherein the effector molecule is a therapeutic agent selected from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid.

19. The isolated anti-RTMC construct of any one of claims 1-10, wherein the isolated anti- RTMC construct is an immunoconjugate comprising the antibody moiety and a label.

20. A host cell expressing the isolated anti-RTMC construct of any one of claims 1-19.

21. A nucleic acid encoding one or more polypeptides contained in the isolated anti-RTMC construct of any one of claims 1-19.

22. A vector comprising the nucleic acid of claim 21.

23. An effector cell expressing the isolated anti-RTMC construct of claim 16 or 17.

24. The effector cell of claim 23, wherein the effector cell is a T cell.

25. A pharmaceutical composition comprising the isolated anti-RTMC construct of any one of claims 1-19, the nucleic acid of claim 21, the vector of claim 22, or the effector cell of claim 23 or 24.

26. A method for detecting a cell presenting a complex comprising an HIV-1 RT peptide and an MHC class I protein on its surface, comprising contacting the cell with the isolated anti- RTMC construct of claim 19 and detecting the presence of the label on the cell.

27. A method for treating an individual having an HIV-1 infection, comprising administering to the individual:

a) an effective amount of the pharmaceutical composition of claim 25, or

b) an effective amount of the effector cell of claim 23 or 24 .

28. A method of diagnosing an individual having an HIV-1 infection, comprising:

a) administering an effective amount of the isolated anti-RTMC construct of claim 19 to the individual; and

29. A method of diagnosing an individual having an HIV-1 infection, comprising:

a) contacting a sample derived from the individual with the isolated anti-RTMC construct of claim 19; and b) determining the number of cells bound with the isolated anti-RTMC construct in the sample, wherein a value for the number of cells bound with the isolated anti-RTMC construct above a threshold level indicates that the individual has the HIV-1 infection.

30. The method of any one of claims 27-29, wherein the individual is human.