CN117683147A

CN117683147A - Chimeric antigen receptor, lentivirus, modified T cell and application thereof

Info

Publication number: CN117683147A
Application number: CN202311766574.5A
Authority: CN
Inventors: 张继帅; 栗红建; 苏红昌; 祖璎玲; 罗燕平; 孙耀威
Original assignee: Shenzhen Prijin Biopharmaceutical Co ltd
Current assignee: Shenzhen Prijin Biopharmaceutical Co ltd
Priority date: 2023-12-20
Filing date: 2023-12-20
Publication date: 2024-03-12

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to a chimeric antigen receptor, a slow virus, a modified T cell and application thereof. The chimeric antigen receptor is obtained by sequentially connecting the following modules in series: signal peptide of CD8 molecule, 5DsdAb single domain antibody, CD8 finger+tm region, 4-1BB cytoplasmic region, cytoplasmic region of CD3 zeta molecule, T2A self-cleaving region, 5DsdAb single domain antibody, ER retention domain. The chimeric antigen receptor can effectively block the expression of CD5 on T cells and prevent the suicide phenomenon of targeted CD5CAR-T cells; the CD5CAR-T cells have obvious specific killing on CCRF-CEM cells, and IFN-gamma secretion level is obviously increased after the CCRF-CEM cells are killed; provides wide application prospect for the curative effect and durability of treating T cell malignant tumor.

Description

Chimeric antigen receptor, lentivirus, modified T cell and application thereof

Technical Field

The invention belongs to the technical field of biological medicine, and particularly relates to a chimeric antigen receptor, a slow virus, a modified T cell and application thereof.

Background

Chimeric antigen receptor (chimeric antigen receptor, CAR) is a fusion protein consisting of an intracellular signal transduction region (e.g. cd3ζ), a transmembrane region and an extracellular antibody single chain variable region fragment (single chain fragment variable, scFV). The specificity of CARs is largely determined by the extracellular scFV region, which is capable of specifically recognizing tumor antigens, signaling antigens, and thereby activating CAR-T cells to exert their cytotoxic effects. The novel accurate targeted therapy for treating tumors has a good effect on clinical tumor treatment through optimization and improvement in recent years, and is a novel tumor immunotherapy method which is very promising, can be accurate, rapid and efficient and can possibly cure cancers.

The CAR-T treatment is also called chimeric antigen receptor T cell treatment, which is to modify human T cells in vitro by genetic engineering means, thereby specifically recognizing and combining antigens on the surface of tumor cells and realizing specific killing of the tumor cells, and reinjecting the tumor cells into a patient for treating diseases.

The production process of CAR-T cells involves the initial isolation and enrichment of T cells, CART cell preparation, including T cell activation, T cell expansion, gene transfer of CAR vectors using viral or non-viral vectors. Lentiviral vectors are considered safe and effective, and they can be randomly integrated into the host T cell genome. Lentiviral expression vectors, known as shuttle vectors, contain the genetic information required for packaging, transfection, stable integration. Lentiviral packaging plasmids can provide all of the helper proteins required to transcribe and package RNA into a recombinant pseudoviral vector. In order to produce high titer viral particles, it is necessary to co-transfect cells with an expression vector and a packaging plasmid, package the virus in the cells, secrete the packaged pseudoviral particles into extracellular medium, centrifuge to obtain supernatant, and then directly use the supernatant for infection of host cells, and after entering the host cells, the target genes are integrated into the genome by reverse transcription, thereby expressing effector molecules at high levels.

The CD5 molecule is a single-chain transmembrane protein comprising an exon region of about 369 amino acid residues, a transmembrane region and an intracellular region of less than about 50 amino acid residues. The exon comprises 3 Ig-like domains, wherein the first domain differs from the other Ig-like domains. CD5 is a pan-T cell marker that plays an important role in T cell development, activation and functional regulation. CD5 is ubiquitously overexpressed in most T cell malignancies, so CD5 molecules are expected to be an ideal target for T cell malignancies. However, both normal effector T cells and T cell tumors express CD5 antigen, which results in the self-phase killing of CD5CAR-T cells, and CD5CAR-T cells are difficult to prepare successfully in vitro. There is therefore a need to develop a chimeric antigen receptor that effectively reduces suicide targeting CD5CAR-T cells.

Disclosure of Invention

In order to solve the problems, the humanized nano antibody is adopted as the recognition domain of the CAR antigen, and the CD5 protein is prevented from being transferred to the cell surface in the process of intracellular expression by a CD5 binding domain endoplasmic reticulum anchoring method, so that the expression of CD5 on the cell surface is effectively reduced, the suicide phenomenon of the targeted CD5CAR-T cell is furthest reduced, and the humanized nano antibody has the advantages of being capable of surviving in vivo for a long time, thereby achieving better curative effect.

In one aspect, the invention provides a chimeric antigen receptor that targets a CD5 molecule, the chimeric antigen receptor comprising a 5DsdAb single domain antibody; the 5Dsdab single domain antibody comprises a heavy chain variable region CDR1, a CDR2 and a CDR3; the amino acid sequences of the CDR1, the CDR2 and the CDR3 are shown as SEQ ID NO.15-SEQ ID NO. 17.

Specifically, the amino acid sequence of the 5Dsdab single domain antibody is SEQ ID No.1 or a sequence with 85% homology with SEQ ID No.1.

Preferably, the amino acid sequence of the 5DsdAb single domain antibody is SEQ ID No.1 or a sequence having 90% homology with SEQ ID No.1.

Further preferably, the amino acid sequence of the 5DsdAb single domain antibody is SEQ ID No.1 or a sequence having 95% homology with SEQ ID No.1.

Further preferably, the amino acid sequence of the 5DsdAb single domain antibody is SEQ ID No.1 or a sequence having 98% homology with SEQ ID No.1.

Still more preferably, the amino acid sequence of the 5DsdAb single domain antibody is SEQ ID No.1.

Specifically, the chimeric antigen receptor also includes, but is not limited to: signal peptide, extracellular region, transmembrane region and/or cytoplasmic region.

Specifically, the cytoplasmic region is selected from at least one of CD8, 4-1BB and CD3 zeta.

Specifically, the chimeric antigen receptor further comprises a self-cleaving region and a retention domain.

Specifically, the chimeric antigen receptor is obtained by sequentially and serially connecting the following modules: signal peptide of CD8 molecule, 5DsdAb single domain antibody, CD8 finger+tm region, 4-1BB cytoplasmic region, cytoplasmic region of CD3 zeta molecule, T2A self-cleaving region, 5DsdAb single domain antibody, ER retention domain.

More specifically, the amino acid sequence of the signal peptide of the CD8 molecule is shown as SEQ ID NO. 2; the amino acid sequence of the CD8 finger+TM region is shown as SEQ ID NO. 3; the amino acid sequence of the 4-1BB cytoplasmic domain is shown as SEQ ID NO. 4; the amino acid sequence of the cytoplasmic region of the CD3 zeta molecule is shown as SEQ ID NO. 5; the amino acid sequence of the T2A self-shearing region is shown as SEQ ID NO. 6; the amino acid sequence of the ER retention domain is shown in SEQ ID NO. 7.

Specifically, the amino acid sequence of the chimeric antigen receptor is shown as SEQ ID NO.9.

In yet another aspect, the invention provides a nucleic acid that expresses the chimeric antigen receptor described above.

Specifically, the nucleic acid sequence is shown as SEQ ID NO.8 or a sequence with more than 80% of sequence homology with SEQ ID NO. 8.

Preferably, the nucleic acid sequence is shown as SEQ ID NO.8 or has more than 85% sequence homology with SEQ ID NO. 8.

Further preferably, the nucleic acid sequence is as shown in SEQ ID NO.8 or a sequence having more than 90% sequence homology with SEQ ID NO. 8.

Still more preferably, the nucleic acid sequence is as shown in SEQ ID NO.8 or a sequence having more than 95% sequence homology with SEQ ID NO. 8.

Most preferably, the nucleic acid sequence is as shown in SEQ ID NO.8 or a sequence having more than 98% sequence homology with SEQ ID NO. 8.

In yet another aspect, the invention provides a lentivirus comprising the chimeric antigen receptor described above or the nucleic acid described above.

In yet another aspect, the invention provides a cell comprising the chimeric antigen receptor described above or the nucleic acid described above or the lentivirus described above.

In particular, the cells include, but are not limited to, immune effector cells.

Further specifically, the immune effector cells include, but are not limited to: t cells, B cells, natural killer cells, macrophages, NKT cells, monocytes, dendritic cells, granulocytes, lymphocytes, leukocytes and/or peripheral blood mononuclear cells.

Still more particularly, the immune effector cell is an engineered immune effector cell.

Preferably, the engineered immune effector cells include, but are not limited to: CAR-T cells or CAR-NK cells.

In yet another aspect, the invention provides a medicament comprising the chimeric antigen receptor described above or the nucleic acid described above or the lentivirus described above or the cell described above.

Specifically, the medicine also comprises pharmaceutically acceptable auxiliary materials.

Preferably, the pharmaceutically acceptable auxiliary materials are selected from polysorbate, histidine, sucrose, arginine, sodium chloride, methionine, acetate, trehalose, proline, sorbitol, sodium phosphate, poloxamer 188, ethylenediamine tetraacetic acid, citric acid, mannitol, glutamate, glycine, sodium citrate, sodium succinate and/or lactic acid.

Specifically, the pharmaceutical dosage forms include, but are not limited to: liquid solution only, lyophilized powder, prefilled syringe only, prefilled syringe, lyophilized powder, tablet, capsule, granule, spray.

In particular, the routes of administration include, but are not limited to: intraocular injection, intravenous injection, intramuscular injection, subcutaneous injection, nasal inhalation, and oral administration.

In a further aspect, the invention provides the use of the chimeric antigen receptor as defined above or the nucleic acid as defined above or the lentivirus as defined above or the cell as defined above for the preparation of a medicament for the treatment of a neoplastic disease.

Specifically, the medicament further comprises one or more other antitumor medicaments.

In particular, the neoplasm may be a T cell malignancy.

In particular, the T cell malignancies include, but are not limited to: leukemia, lymphoma.

In a further aspect, the invention provides the use of the chimeric antigen receptor as defined above or the nucleic acid as defined above or the lentivirus as defined above or the cell as defined above in the manufacture of a medicament for the treatment of an autoimmune disease.

The invention has the technical effects that:

(1) The 5D-ER can effectively block the expression of CD5 on T cells and prevent suicide phenomenon of targeted CD5 CART cells;

(2) Under the condition of different effective target ratios, the CD5CAR-T cells have obvious specific killing on CCRF-CEM cells;

(3) IFN-gamma secretion levels were significantly increased after 5 D.b. CAR-T killing of CCRF-CEM cells.

Drawings

FIG. 1 is a schematic diagram of CD5-CART structure with CD5 blocking; wherein CD8SP represents a signal peptide of a CD8 molecule; the 5DsdAb represents a single domain antibody sequence that targets CD 5; CD8 finger+TM represents the extracellular region and transmembrane region sequence of the CD8 molecule; 4-1BB represents a 4-1BB cytoplasmic domain sequence; cd3ζ represents the cytoplasmic sequence of the cd3ζ molecule; T2A represents a self-cleaving sequence; ER represents ER retention domain sequences.

FIG. 2 is a map of lentiviral vector Pre-Lenti-EF1-CAR V2 Wpmut.

FIG. 3 is a map of packaging plasmid pMDLg-pRRE-K.

FIG. 4 is a map of the packaging plasmid pRsv-rev-kana-V2.

FIG. 5 is a map of the envelope plasmid pMD2. G-K.

FIG. 6 is a flow assay for CD5CAR-T CAR positive rate of CD5 expression block and CD5 molecule expression on cell membrane; wherein, A is 5D.b CAR-T cell CAR positive rate is close to 100%; b is 5DER effectively blocks CD5 expression on T cells, whereas the positive rate of CD5 on the surface of T cells without transfection of CD5 blocking virus is 99.5%.

FIG. 7 is cytotoxicity of CD5CAR-T cells with blocked CD5 expression on CD 5-positive acute lymphoblastic leukemia cells (CCRF-CEM).

FIG. 8 shows cytokine IFN-gamma secretion following incubation of CD5CAR-T with CCRF-CEM cells with blocked CD5 expression.

Detailed Description

The present invention will be described in further detail with reference to the following examples, which are not intended to limit the present invention, but are merely illustrative of the present invention. The experimental methods used in the following examples are not specifically described, but the experimental methods in which specific conditions are not specified in the examples are generally carried out under conventional conditions, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.

EXAMPLE 1 construction of CD5-CART vector with CD5 molecular blocking

The embodiment constructs a CAR lentiviral vector with an intrablock CD5 expression blocking function and targeting CD5, the structure is shown in figure 1, a 5D single domain antibody is taken as an antigen recognition region, a CD8hinge region and CD8 transmembrane region sequence, a 4-1BB intracellular domain sequence and an intracellular signal molecule CD3 zeta sequence are combined, the CD5 single domain antibody sequence is connected through T2A, and the ER retention domain is connected, so that the CAR structure of targeting CD5 of T cells is named as 5D.b CAR. The lentiviral vector Pre-Lenti-EF1-CAR V2Wpmut and 5D.b CAR genes are subjected to enzyme digestion, connection, transformation, cloning, plasmid extraction and sequencing, and the lentiviral vector Pre-Lenti-EF1-5DER-CAR V2WPmut with the correct sequence is obtained. The nucleotide sequence of the 5D.b CAR is SEQ ID NO.8, and the corresponding amino acid sequence is SEQ ID NO.9.

The preparation method of lentiviral vector Pre-Lenti-EF1-CAR V2Wpmut (shuttle plasmid) is as follows:

(1) The 5'LTR of the shuttle plasmid pRRLSIN.cPPT.PGK-GFP.WPRE (http:// n2 t.net/adedge: 12252) is taken as an RSV promoter, and the promoter of the 5' LTR is changed from RSV to CMV, so that the efficient transcription of the viral genome sequence is facilitated; the 3' LTR original plasmid has removed the U3 region (3 ' LTR. DELTA.U3), has been designed for self-inactivation (SIN), and the 3' LTR sequence is unchanged.

(2) The hGGK promoter driving the expression of exogenous transgene is replaced by EF1 promoter, EGFP gene is replaced by polyclonal sequence (SEQ ID NO. 10);

(3) The ampicillin resistance gene was replaced with a kana resistance gene derived from pUC57-Kan (GenBank: LT 671993.1) plasmid;

(4) The WPRE element of pRRLSIN.cPPT.PGK-GFP.WPRE was modified so that truncated X protein was not expressed. The method comprises the following steps: within the sequence of the WPRE wild-type element, the promoter driving the transcription of protein X was deleted, the a of the initiation codon ATG was changed to T, and the new sequence was named WPREmut.

The modified shuttle plasmid is named as Pre-Lenti-EF1-CAR V2Wpmut, and the sequence is shown in SEQ ID NO. 11. The shuttle plasmid map is shown in FIG. 2, annotated in the map is shown in Table 1 below.

TABLE 1 shuttle plasmid Pre-Lenti-EF1-CAR V2WPmut elements annotation

EXAMPLE 2 lentiviral packaging

2.1 preparation of packaging plasmid pMDLg-pRRE-K

The packaging plasmid is derived from a plasmid pMDLg-pRRE, is initially constructed by Dier Trono Lab, is stored in Addgene (www.addgene.org/12251 /), has ampicillin resistance in a primary sequence, and is replaced by a kana resistance gene of a pET-28 (a) plasmid; the modified packaging plasmid is named pMDLg-pRRE-K, and the sequence is shown in SEQ ID NO. 12. The map of packaging plasmid pMDLg-pRRE-K is shown in FIG. 3 and the annotation is shown in Table 2.

TABLE 2

Element name	Functional area
		CMV promoter	Human cytomegalovirus promoter
β-globin intron	Human beta globulin truncated intron fragment
		HIV-1gag	Encoding gag protein
HIV-1pol	Encoding pol proteins
		RRE	Rev protein response element of HIV-1
β-globin poly(A)	Human beta-globulin polyadenylation signal
		Ori	ColE1/pMB1/pBR322 high copy replication origin
KanR	Kanamycin resistance gene

2.2 preparation of packaging plasmid pRsv-rev-kana-V2

The packaging plasmid is derived from plasmid pRSV-rev, is initially constructed by Dier Trono Lab, is stored in Addgene (www.addgene.org/12253 /), has ampicillin resistance in the original sequence, and is replaced by a kana resistance gene of pUC57-Kan (GenBank: LT 671993.1) plasmid; the above modified packaging plasmid was designated pRsv-rev-kana-V2 and the sequence shown in SEQ ID No. 13. The map of the packaging plasmid pRsv-rev-kana-V2 is shown in FIG. 4 and annotated as Table 3 below.

TABLE 3 Table 3

Element name	Functional area
		RSV promoter	RSV promoters
Rev	Viral gene expression modulators
		Ori	ColE1/pMB1/pBR322 high copy replication origin
KanR	Kanamycin resistance gene

2.3 preparation of the envelope plasmid pMD2.G-K

(1) The envelope plasmid pMD2.G, originally constructed from Dier Trono Lab, was stored in Addgene, (https:// www.addgene.org/12259 /). Based on its sequence and backbone, replacing its ampicillin resistance gene with the kana resistance gene of pET-28 (a) plasmid;

(2) The modified envelope plasmid is named pMD2.G-K, and the sequence is shown as SEQ ID NO. 14.

(3) The map of the envelope plasmid is shown in FIG. 5, and the annotation is shown in Table 4 below.

TABLE 4 Table 4

Element name	Functional area
		CMV enhancer	Human cytomegalovirus enhancer
CMV promoter	Human cytomegalovirus promoter
		β-globin intron	Human beta-globulin truncated intronFragments
VSV-G	Vesicular stomatitis virus G glycoprotein
		β-globin poly(A)	Human beta-globulin polyadenylation signal
Ori	ColE1/pMB1/pBR322 high copy replication origin
		KanR	Kanamycin resistance gene

2.4 preparation of lentiviruses

And adopting a three-generation vector and four-plasmid system for packaging. The Pre-Lenti-EF1-5D-ER-CAR V2WPmut CAR shuttle plasmid was mixed with the packaging plasmid pMDLg-pRRE-K, the envelope plasmid pMD2.G-K, the packaging plasmid pRsv-rev-kana-V2 in a ratio of 7:5:3:5, and the mixture of the above four plasmids was added to a tube containing 293TS basal medium. According to the plasmid: PEIpro = 1:2 ratio to another tube. After the two tubes are incubated at room temperature for 5min, the mixed solution containing PEIpro is slowly added into the mixed solution containing DNA, the centrifuge tube is gently rocked to mix uniformly, and the tubes are incubated at room temperature for 15min. The transfection mixture was then added drop-wise to a 293TS (ATCC, cat# CRL-3216) cell suspension. After 48 hours, collecting culture medium supernatant to obtain slow virus crude liquid. Then, lentivirus was concentrated, and the virus was stored at 80℃after split charging. Transduction titers of lentiviruses were determined based on flow cytometry.

The detection of transduction titres comprises the following steps:

the lentivirus crude venom was serially diluted and then infected with 293T cells, and after about 70h of infection, the sample CAR positive rate was detected using flow cytometry, transduction titer (TU/mL) =dilution multiple sample positive rate cell amount/virus plusSample size (mL). Transduction titres 3.51X10 ⁷ TU/mL。

Example 3 preparation of CD5CAR-T cells with blocked CD5 expression

Peripheral blood was collected from healthy volunteers and purified using Miltenyi CD3 Regent to obtain cd3+ T cells. Cells were grown in TexMACS GMP Medium (5% serum replacement) medium and T cells were activated using the transact CD3/CD28 reagent (1:100) and IL-2 (100 IU/ml). After 24 hours of T cell activation, the cells were infected with lentiviruses.

Example 4CD5 expression blocked CD5CAR-T CAR Positive Rate and expression of CD5 molecules on cell membranes

Primary T cells were transduced by lentiviral vectors and the CD5CAR-T CAR positive rate and CD5 molecule expression on the cell membrane were detected using flow-through. The results are shown in fig. 6, which shows that the 5d.b CAR-T cell CAR positive rate is close to 100%, (a in fig. 6). The 5DER (5D single domain antibody sequence and attached ER retention domain) effectively blocked CD5 expression on T cells, whereas the positive rate of CD5 on the T cell surface without transfected CD5 blocking virus was 99.5% (shown as B in fig. 6), suggesting that the 5DER could bind to CD5 in the cell and retain it in the endoplasmic reticulum. Therefore, this CD5 expression blocking technique can be used to block CD5 expression on cells, preventing suicide phenomena targeting CD5 CART cells.

Example 5 in vitro killing of CD 5-positive tumor cells by CD5CAR-T cells with blocked CD5 expression

Cytotoxicity of CD 5-positive acute lymphoblastic leukemia cells (CCRF-CEM) by CD5CAR-T cells blocked by 5.1CD5 expression

CD5CAR-T cells with blocked CD5 expression an in vitro killing experiment was performed on acute lymphoblastic leukemia cells (CCRF-CEM), incubated with CCRF-CEM cells for 16 hours at a 4:1, 2:1, 1:1 target-to-target ratio, respectively, and LDH release experiments assessed the killing function of 5d.b CAR-T cells. The results are shown in figure 7, where CD5CAR-T cells had significant specific killing of CCRF-CEM cells compared to control under conditions of different effective target ratios.

Cytokine secretion after incubation of 5.2CD5 expression blocked CD5CAR-T with CCRF-CEM cells

The killed supernatant was collected and assayed for IFN-gamma secretion following killing of CCRF-CEM cells by 5D.b CAR-T cells by ELISA. The results are shown in figure 8, where T cells were substantially free of secretion of the cytokine IFN- γ after co-incubation with CCRF-CEM cells, whereas the level of IFN- γ secretion was significantly increased after 5d.b CAR-T killing of CCRF-CEM cells.

Claims

1. A chimeric antigen receptor that targets a CD5 molecule, wherein said chimeric antigen receptor comprises a 5DsdAb single domain antibody; the 5Dsdab single domain antibody comprises a heavy chain variable region CDR1, a CDR2 and a CDR3; the amino acid sequences of the CDR1, the CDR2 and the CDR3 are shown as SEQ ID NO.15-SEQ ID NO. 17.

2. The chimeric antigen receptor according to claim 1, wherein the amino acid sequence of the 5DsdAb single domain antibody is SEQ ID No.1 or a sequence having 85% homology with SEQ ID No.1.

3. The chimeric antigen receptor according to claim 1, wherein the chimeric antigen receptor further comprises a signal peptide, an extracellular region, a transmembrane region, and/or a cytoplasmic region.

4. The chimeric antigen receptor according to claim 3, wherein the cytoplasmic domain is selected from at least one of CD8, 4-1BB and cd3ζ.

5. The chimeric antigen receptor according to claim 1, further comprising a self-cleaving region and a retention domain.

6. The chimeric antigen receptor according to any one of claims 1-5, wherein the chimeric antigen receptor is obtained by sequential tandem connection of: signal peptide of CD8 molecule, 5DsdAb single domain antibody, CD8 finger+tm region, 4-1BB cytoplasmic region, cytoplasmic region of CD3 zeta molecule, T2A self-cleaving region, 5DsdAb single domain antibody, ER retention domain.

7. The chimeric antigen receptor according to claim 6, wherein the signal peptide of the CD8 molecule has the amino acid sequence shown in SEQ ID No. 2; the amino acid sequence of the CD8 finger+TM region is shown as SEQ ID NO. 3; the amino acid sequence of the 4-1BB cytoplasmic domain is shown as SEQ ID NO. 4; the amino acid sequence of the cytoplasmic region of the CD3 zeta molecule is shown as SEQ ID NO. 5; the amino acid sequence of the T2A self-shearing region is shown as SEQ ID NO. 6; the amino acid sequence of the ER retention domain is shown in SEQ ID NO. 7.

8. The chimeric antigen receptor according to claim 7, wherein the amino acid sequence of the chimeric antigen receptor is shown in SEQ ID No.9.

9. A nucleic acid expressing the chimeric antigen receptor of any one of claims 1-8.

10. The nucleic acid of claim 9, wherein the nucleic acid has a sequence as shown in SEQ ID No.8 or a sequence having 80% or more sequence homology with SEQ ID No. 8.

11. A lentivirus comprising the chimeric antigen receptor of any one of claims 1-8 or the nucleic acid of any one of claims 9-10.

12. A cell comprising the chimeric antigen receptor of any one of claims 1-8 or the nucleic acid of any one of claims 9-10 or the lentivirus of claim 11.

13. The cell of claim 12, wherein the cell comprises an immune effector cell.

14. The cell of claim 13, wherein the immune effector cell comprises a T cell, B cell, natural killer cell, macrophage, NKT cell, monocyte, dendritic cell, granulocyte, lymphocyte, leukocyte, and/or peripheral blood mononuclear cell.

15. The cell of claim 14, wherein the immune effector cell is an engineered immune effector cell.

16. The cell of claim 15, wherein the engineered immune effector cell comprises a CAR-T cell or a CAR-NK cell.

17. A medicament comprising the chimeric antigen receptor of any one of claims 1 to 8 or the nucleic acid of any one of claims 9 to 10 or the lentivirus of claim 11 or the cell of any one of claims 12 to 16.

18. The medicament of claim 17, further comprising pharmaceutically acceptable excipients.

19. Use of the chimeric antigen receptor of any one of claims 1-8 or the nucleic acid of any one of claims 9-10 or the lentivirus of claim 11 or the cell of any one of claims 12-16 in the preparation of a medicament for treating a neoplastic disease.

20. Use of the chimeric antigen receptor of any one of claims 1-8 or the nucleic acid of any one of claims 9-10 or the lentivirus of claim 11 or the cell of any one of claims 12-16 in the manufacture of a medicament for the treatment of an autoimmune disease.