BR122024005916A2 - USE OF AN ANTIBODY THAT SPECIFICALLY BINDS TO COMPONENT 1S (C1S) OF COMPLEMENT IN THE TREATMENT OF AUTOIMMUNE DISORDERS - Google Patents

Abstract

The present disclosure provides methods of treating an alloimmune or autoimmune disorder in an individual; The methods involve administering to the individual an effective amount of an antibody specific to the C1s component of complement. The present disclosure provides a method of monitoring the effectiveness of a method of treating a subject; The method involves detecting the level of autoantibodies or alloantibodies in a biological sample obtained from the individual.

Description

CROSS REFERENCE

[001] This application claims the benefit of Provisional Patent Application No.] US 62/185,362, filed on June 26, 2015, an application that is incorporated into this document by reference in its entirety.

INTRODUCTION

[002] The complement system is a known effector mechanism of the immune response, providing not only protection against pathogens and other harmful agents, but also recovery from injuries. The complement pathway comprises numerous proteins that typically exist in the body in inactive form. The classical complement pathway is triggered by the activation of the first component of complement, known as the C1 complex, which consists of the proteins C1q, C1r, and C1s. Upon binding of C1 to an immune complex or other activator, the C1s component, a serine protease sensitive to diisopropyl fluorophosphate (DFP), cleaves complement components C4 and C2 to initiate activation of the classical complement pathway. complement. The classical complement pathway appears to play a role in many diseases and disorders, including autoimmune disorders and alloimmune disorders.

[003] There is a need in the art for compounds that treat a complement-mediated disease or disorder.

SUMMARY

[004] The present invention provides methods of treating an alloimmune or autoimmune disorder in an individual; The methods involve administering to the individual an effective amount of an antibody specific to the C1s component of complement. The present invention provides a method of monitoring the effectiveness of a method of treating a subject; The method involves detecting the level of autoantibodies or alloantibodies in a biological sample obtained from the individual.

BRIEF DESCRIPTION OF FIGURES

[005] FIG. 1 represents the amino acid sequence of the Homo sapiens complement C1s protein (SEQ ID NO:158).

[006] FIGS. 2A-2D depict the effect of TNT003 on the activation of normal primary B cells induced by B cell receptor agonists in the presence of normal human serum.

[007] FIGS. 3A-3C represent the effect of a humanized variant of TNT003 on the activation and proliferation of normal primary human B cells induced by a B cell receptor agonist in the presence of normal human serum.

[008] FIGS. 4A-4C represent the effect of C1s inhibitor antibody (a humanized variant of TNT003) and C5 inhibitor on the activation of normal primary human B cells induced by a B cell receptor agonist in the presence of normal human serum.

[009] FIGS. 5A-5C represent the effect of several C1s inhibitory antibodies and a C5 inhibitory antibody on the activation of primary human B cells induced by a B cell receptor agonist in the presence of normal human serum.

DEFINITIONS

[0010] The terms "antibodies" and "immunoglobulin" include antibodies or immunoglobulins of any isotype, antibody fragments that retain specific binding to the antigen, including, but not limited to, Fab, Fv, scFv and Fd fragments, chimeric antibodies, humanized antibodies, single-chain antibodies (scAb), single-domain antibodies (dAb), single-domain heavy-chain antibodies, single-domain light-chain antibodies, bispecific antibodies, multispecific antibodies, and fusion proteins comprising a moiety antigen binding (also referred to herein as antigen binding) of an antibody and a protein other than the antibody. Antibodies can be detectably labeled, for example, with a radioisotope, an enzyme that generates a detectable product, a fluorescent protein, and the like. Antibodies can be further conjugated with other moieties, such as members of specific binding pairs, for example, biotin (biotin-avidin specific binding pair member) and the like. Antibodies may also be attached to a solid support, including, but not limited to, polystyrene plates or beads and the like. Also covered by the term are Fab', Fv, F(ab')2 and/or other antibody fragments that retain specific binding to the antigen and monoclonal antibodies. As used herein, a monoclonal antibody is an antibody produced by a group of identical cells, all of which were produced from a single cell by repetitive cellular replication. In other words, the cell clone produces only a single species of antibody. Although a monoclonal antibody can be produced using hybridoma production technology, other production methods known to those skilled in the art can also be used (e.g., antibodies derived from antibody phage display libraries). An antibody can be monovalent or bivalent. An antibody can be an Ig monomer, which is a "Y-shaped" molecule consisting of four polypeptide chains: two heavy chains and two light chains linked by disulfide bonds. An antibody may comprise heavy and/or light chain constant regions of any isotype; for example, an antibody may be an IgG1, IgG2a, IgG2b, IgG3 or IgG4 and may have lambda or kappa light chains. The constant region of the heavy chain may be a variant with altered (e.g., increased) binding to an Fc receptor (e.g., FcRn).

[0011] The term "humanized immunoglobulin", as used herein, refers to an immunoglobulin comprising portions of immunoglobulins of different origin, wherein at least one portion comprises amino acid sequences of human origin. For example, the humanized antibody may comprise portions derived from an immunoglobulin of non-human origin with the required specificity, such as from a mouse, and from immunoglobulin sequences of human origin (e.g., chimeric immunoglobulin) chemically joined by conventional techniques (e.g., example, synthetic) or prepared as a contiguous polypeptide using genetic engineering techniques (e.g., DNA encoding portions of chimeric antibody proteins can be expressed to produce a contiguous polypeptide chain). Another example of humanized immunoglobulin is an immunoglobulin containing one or more immunoglobulin chains comprising a CDR derived from an antibody of non-human origin and a framework region derived from a light and/or heavy chain of human origin (e.g. CDR-grafted antibodies with or without framework changes). Chimeric or CDR-grafted single-chain antibodies are also encompassed by the term humanized immunoglobulin. See, for example, Cabilly et al., Pat. No. US 4,816,567; Cabilly et al., European Patent No. 0,125,023 B1; Boss et al., Pat. No. US 4,816,397; Boss et al., European Patent No. 0,120,694 B1; Neuberger, M.S. et al., WO 86/01533; Neuberger, M. S. et al., European Patent No. 0,194,276 B1; Winter, Pat. No. US 5,225,539; Winter, European Patent No. 0,239,400 B1; Padlan, EA et al., European Patent Application No. 0,519,596 A1. See also, Ladner et al., Pat. No. US 4,946,778; Houston, Pat. No. 5,476,786; and Bird, R. E. et al., Science, 242: 423-426 (1988)), in relation to single-chain antibodies.

[0012] For example, humanized immunoglobulins can be produced using synthetic and/or recombinant nucleic acids to prepare genes (e.g., cDNA) that encode the desired humanized chain. For example, nucleic acid sequences (e.g., DNA) encoding humanized variable regions can be constructed using PCR mutagenesis methods to alter DNA sequences encoding a human or humanized strand, such as a DNA template of a variable region. previously humanized (see, for example, Kamman, M., et al., Nucl. Acids Res., 17: 5404 (1989)); Sato, K., et al., Cancer Research, 53: 851-856 (1993); Daugherty, B. L. et al., Nucleic Acids Res., 19(9): 2471-2476 (1991); and Lewis, A. P. and J. S. Crowe, Gene, 101: 297-302 (1991)). By using these or other suitable methods, variants can also be readily produced. For example, cloned variable regions can be mutagenized and sequences encoding variants with the desired specificity can be selected (e.g., from a phage library, see, e.g., Krebber et al., Pat. No. US 5,514,548; Hoogenboom et al., WO 93/06213, published April 1, 1993)).

[0013] "Antibody fragments" comprise a portion of an intact antibody, for example, the antigen-binding region or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2 and Fv fragments; diabodies; linear antibodies (Zapata et al, Protein Eng. 8(10): 1057-1062 (1995)); domain antibodies (dAb; Holt et al. (2003) Trends Biotechnol. 21:484); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Digestion of antibodies with papain produces two identical antigen-binding fragments called "Fab" fragments, each with a unique antigen-binding site, and a residual "Fc" fragment, a designation that reflects the ability to crystallize readily. Pepsin treatment yields an F(ab')2 fragment that has two antigen-combining sites and is further capable of cross-linking antigens.

[0014] "Fv" is the minimal antibody fragment that contains a complete antigen binding and recognition site. This region consists of a dimer of a light chain variable domain and a heavy chain variable domain in close non-covalent association. It is in this configuration that the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three antigen-specific CDRs) has the ability to recognize and bind the antigen, albeit at a lower affinity compared to the entire binding site.

[0015] The "Fab" fragment also contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab fragments differ from Fab' fragments by the addition of some residues at the carboxy terminal end of the CH1 domain of the heavy chain including one or more cysteines of the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine residue(s) of the constant domains contain a free thiol group. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments, which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0016] The "light chains" of antibodies (immunoglobulins) of any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. There are five main classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM and several of these classes can be further divided into subclasses (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA and IgA2. Subclasses can be further divided into types, for example, IgG2a and IgG2b.

[0017] "Single-chain Fv" or "sFv" or "scFv" antibody fragments comprise the VH and VL domains of the antibody, where these domains are present in a simple polypeptide chain. In some embodiments, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the sFv to form the desired structure for binding to the antigen. 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).

[0018] The term "diabodies" refers to small antibody fragments with two antigen-binding sites whose fragments comprise a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain. ca (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with complementary domains on another chain and create two antigen-binding sites. Diabodies are more fully described in, for example, EP 404,097; WO 93/11161; and Hollinger et al. (1993) Proc. Natl. Academic. Sci. USA 90:6444-6448.

[0019] As used herein, the term "affinity" refers to the equilibrium constant for the reversible binding of two agents (e.g., an antibody and an antigen) and is expressed as a dissociation constant (KD). The affinity can be at least 1 times greater, at least 2 times greater, at least 3 times greater, at least 4 times greater, at least 5 times greater, at least 6 times greater, at least 7 times greater, at least 8 times greater greater, at least 9 times greater, at least 10 times greater, at least 20 times greater, at least 30 times greater, at least 40 times greater, at least 50 times greater, at least 60 times greater, at least 70 times greater, at least 80 times greater, at least 90 times greater, at least 100 times greater, or at least 1000 times greater or more than the affinity of an antibody for unrelated amino acid sequences. The affinity of an antibody for a target protein may be, for example, from about 100 nanomolar (nM) to about 0.1 nM, from about 100 nM to about 1 picomolar (pM), or from about 100 nM to about 1 femtomolar (fM) or more. As used herein, the term "avidity" refers to the resistance of a complex of two or more agents to dissociation upon dilution. The terms "immunoreactive" and "preferentially binds" are used interchangeably herein with respect to antibody and/or antigen-binding fragments.

[0020] The term "bond" refers to a direct association between two molecules, due, for example, to covalent, electrostatic, hydrophobic and ionic and/or hydrogen bonding interactions, including interactions such as salt bridges and water bridges . A subject's anti-C1s antibody specifically binds to an epitope within a complement C1s protein. "Specific binding" refers to binding with an affinity of at least about 10-7 M or greater, e.g., 5 x 10-7 M, 10-8 M, 5 x 10-8 M and greater. "Non-specific binding" refers to a binding with an affinity of less than about 10-7 M, for example, a binding with an affinity of 10-6 M, 10-5 M, 10-4 M, etc.

[0021] 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 light and heavy chain polypeptides. CDRs were described by Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., US Dept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991) (also referred to herein as Kabat 1991); by Chothia et al., J. Mol. Biol. 196:901-917 (1987) (also referred to here as Chothia 1987); and MacCallum et al., J. Mol. Biol. 262:732-745 (1996), where definitions include overlapping or subsets of amino acid residues compared to each other. However, application of any definition to refer to a CDR of a grafted antibody or 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 references cited above, are presented below in Table 1 as a comparison. The CDRs listed in Table 2 were defined according to Kabat 1991. Table 1: CDR Definitions 1 Residue numbering follows the nomenclature of Kabat et al., supra 2 Residue numbering follows the nomenclature of Chothia et al., supra 3 Residue numbering follows the nomenclature of MacCallum et al., supra

[0022] As used herein, the terms "CDR-L1", "CDR-L2" and "CDR-L3" refer, respectively, to the first, second and third CDRs in a light chain variable region. As used herein, the terms "CDR-H1", "CDR-H2" and "CDR-H3" refer, respectively, to the first, second and third CDRs in a heavy chain variable region. As used herein, the terms "CDR-1", "CDR-2" and "CDR-3" refer, respectively, to the first, second and third CDRs of the variable region of any of the chains.

[0023] As used herein, the term "framework", when used in reference to a variable region of the antibody, is intended to mean all amino acid residues outside the CDR regions within the variable region of an antibody. A variable region framework is generally a discontinuous amino acid sequence between about 100-120 amino acids in length, but is intended to reference only those amino acids outside of the CDRs. As used in this document, the term "framework region" is intended to mean each domain of the framework that is separated by CDRs.

[0024] An "isolated" antibody is an antibody that has been identified and separated and/or recovered from a component of its natural environment. Contaminating components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other protein or non-protein solutes. In some embodiments, the antibody will be purified (1) to greater than 90%, greater than 95%, or greater than 98% by weight of antibody as determined by the Lowry method, e.g., greater than 99% by weight. , (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence through the use of a rotating cup sequencer, or (3) to homogeneity by polyacrylamide-dodecyl sulfate gel electrophoresis (SDS-PAGE) under reducing or non-reducing conditions using Coomassie blue or silver dye. Isolated antibody includes antibody in situ within recombinant cells, since at least one component of the antibody's natural environment will not be present. In some cases, the isolated antibody will be prepared by at least one purification step.

[0025] The terms "polypeptide", "peptide" and "protein", used interchangeably herein, refer to a polymeric form of amino acids of any length, which may include genetically encoded and non-genetically encoded amino acids, chemically or biochemically modified or derivatized amino acids and polypeptides with modified peptide backbones. The term includes fusion proteins, including, but not limited to, fusion proteins with a heterologous amino acid sequence, fusions with heterologous and homologous leader sequences, with or without N-terminal methionine residues; immunologically marked proteins; and the like.

[0026] As used herein, the terms "treatment", "treat" and the like refer to obtaining a desired physiological and/or pharmacological effect. The effect may be prophylactic in terms of complete or partial prevention of a disease or symptom thereof and/or may be therapeutic in terms of a partial or complete cure of a disease and/or adverse effect attributed to the disease. "Treatment", as used herein, encompasses any treatment of a disease in a mammal, particularly a human, and includes: (a) preventing the disease from occurring in a subject who may be predisposed to the disease, but who still have not been diagnosed as having it; (b) inhibit the disease, that is, stop its development; and (c) alleviate the disease, that is, cause regression of the disease.

[0027] The terms "individual", "subject," "host" and "patient", used interchangeably herein, refer to a mammal, including, but not limited to, murine (rats, mice), primates non-humans, humans, canines, felines, ungulates (e.g. horses, cattle, sheep, pigs, goats), etc. Also covered by these terms are any animals that have a complement system, such as mammals, fish, and some invertebrates. As such, these terms include mammals, domestic fish and invertebrates, agricultural animals, working animals, zoo animals, and laboratory animals that contain the complement system.

[0028] A "therapeutically effective amount" or "effective amount" refers to the amount of a C1s anticomplement antibody that, when administered to a mammal or other subject to treat a disease, is sufficient to effect such treatment for the disease. The "therapeutically effective amount" will vary depending on the C1s anticomplement antibody, the disease and its severity, and the age, weight, etc., of the subject being treated.

[0029] A "biological sample" encompasses a variety of sample types obtained from an individual and can be used in a diagnostic or monitoring assay. The definition encompasses samples of blood and other liquids of biological origin, solid tissue samples such as a biopsy specimen or tissue cultures or cells derived therefrom, and their progeny. The definition also includes samples that have been manipulated in any way after their acquisition, such as by treatment with reagents, solubilization, or enrichment for certain components, such as polynucleotides. The term "biological sample" encompasses a clinical sample and also includes cultured cells, cell supernatants, cell lysates, serum, plasma, biological fluid and tissue samples. The term "biological sample" includes urine, saliva, cerebrospinal fluid, interstitial fluid, ocular fluid, synovial fluid, blood fractions such as plasma and serum, and the like. The term "biological sample" also includes solid tissue samples, tissue culture samples, and cellular samples.

[0030] Before the present invention is described in more detail, it should be understood that this invention is not limited to particular described embodiments, since they may, of course, vary. It should also be understood that the terminology used herein is intended to describe particular embodiments only, and is not intended to be limiting, as the scope of the present invention will be limited only by the appended claims.

[0031] Where a range of values is provided, it is understood that each intervening value, up to the tenth of a unit of the lower limit, unless the context clearly dictates otherwise, between the upper and lower limits of the range and any another value established or intervening in this established range is encompassed by the invention. The upper and lower limits of these smaller ranges can be independently included in the smaller ranges and are also encompassed in the invention, subject to any limit specifically excluded in the established range. Where the established range includes one or both limits, ranges that exclude either or both of the included limits are also included in the invention.

[0032] Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as commonly understood by someone skilled in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention, preferred methods and materials will now be described. All publications referenced herein are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited.

[0033] It should be noted that, as used herein and in the attached claims, the singular forms "a", "an", and "the" include plural referents, unless the context clearly dictates otherwise . Thus, for example, reference to "an anti-C1s antibody" includes a plurality of such antibodies and reference to "autoimmune disorder" includes reference to one or more autoimmune disorders and equivalents thereof known to those skilled in the art, and so onwards. It is further noted that claims may be written to exclude any optional elements. As such, this statement is intended to serve as the antecedent basis for the use of such exclusive terminology as "solely," "only" and the like in connection with the recitation of claim elements, or use of a "negative" limitation.

[0034] It is recognized that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. On the other hand, various features of the invention that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination. All combinations of embodiments related to the invention are specifically encompassed by the present invention and are disclosed herein as if each and every combination had been individually and explicitly disclosed. Furthermore, all subcombinations of the various embodiments and elements thereof are also specifically encompassed by the present invention and are disclosed herein as if each such subcombination had been individually and explicitly disclosed herein.

[0035] The publications discussed in this document are provided solely for your description prior to the filing date of this application. Nothing herein should be construed as an admission that the present invention is not entitled to precede such publication by virtue of prior invention. Additionally, the publication dates provided may differ from the actual publication dates which may need to be independently confirmed.

DETAILED DESCRIPTION

[0036] The present invention provides methods of treating an alloimmune or autoimmune disease in an individual; The methods involve administering to the individual an effective amount of an antibody specific to the C1s component of complement in an amount and for a period of time effective to reduce the level of autoantibody or alloantibody titers. The present invention provides a method of monitoring the effectiveness of a method of treating a subject; The method involves detecting the level of autoantibodies or alloantibodies in a biological sample obtained from the individual.

TREATMENT METHODS

[0037] The present invention provides methods of treating an alloimmune or autoimmune disease in an individual. The methods comprise administering to the subject an effective amount of an antibody specific for the C1s complement component. Anti-C1s antibody is administered in an amount and for a period of time effective to reduce the level of autoantibody or alloantibody titers. Administration of anti-C1s antibody is effective in reducing the level of autoantibodies or alloantibodies in the individual.

Reducing the level of autoimmune antibodies

[0038] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and over a period of time to an individual with an autoimmune disorder, is effective in reducing the level of autoantibodies in the individual by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or more than 90%, compared to the level of autoantibodies in the individual in the absence of anti-C1s antibody treatment or compared to the level of autoantibodies in the individual before treatment with the anti-C1s antibody.

[0039] Autoantibodies include, for example, an anti-nuclear antibody, an anti-neutrophil antibody, an anti-ribonucleic protein antibody, an anti-single-stranded DNA antibody, an anti-La/SSA antibody, an anti-La/SS antibody -B, an anticentromere antibody, an antineuronal nuclear antibody-2, an anti-double-stranded DNA antibody, an anti-Jol antibody (where the autoantigen is histidine-tRNA ligase), an anti-Smith antibody (where the autoantigen is a snRNP core protein), an anti-topoisomerase antibody, an anti-histone antibody, an anti-p62 antibody (where the autoantigen is Nucleoporin 62), an anti-sp100 antibody (where the autoantigen is sp100 nuclear antigen), an antitransglutaminase antibody, an anti-ganglioside antibody, an anti-thrombin antibody, an anti-actin antibody, an anti-neutrophil cytoplasmic antibody, an anti-signal recognition particle antibody, an anti-DNA antibody, an anti-Rho antibody, an anti-collagen antibody, an anti-1-antigen antibody, an anti-i-antigen antibody, an anti-collagen XVII antibody, an anti-Rho/SSA antibody, an antiphospholipid antibody, an anti-smooth muscle (anti-Sm) antibody, an anti-mitochondrial antibody, an anti-acetylcholine receptor antibody, an antibody to histidyl tRNA synthetase (HisRS), an anti-voltage-gated calcium channel antibody, an anti-voltage-gated potassium channel antibody, an anti-glycoprotein IIb/IIIa antibody, an anti-glycoprotein Ib/IX antibody, cold agglutinins (e.g., the antibody that binds to a red blood cell, such as an anti-antigen-1 antibody, an anti-antigen-i antibody, an anti-antigen-Pr antibody, etc.), an anti-aquaporin 4 antibody, an anti-muscle-specific kinase (MuSK) antibody and the like. Autoantibodies include antibodies against self-antigens such as myelin basic protein, collagen (e.g., type XI collagen, type XVII collagen), human cartilage gp 39, chromogranin A, gp130- RAPS, proteolipid protein, fibrillarin, Rho autoantigen, l-antigen, i-antigen, Pr antigen, nucleolar proteins (e.g., small nucleolar protein), thyroid-stimulating factor receptor, histones, gp 70 glycoprotein, ribosomal proteins, pyruvate dehydrogenase dehydrolipoamide acetyltransferase, antigens hair follicle, IgG, human tropomyosin isoform 5, mitochondrial proteins, pancreatic β-cell proteins, myelin oligodendrocyte glycoprotein, insulin, glutamic acid decarboxylase (GAD), gluten, acetylcholine receptors, aquaporin 4, muscle-specific kinase (MuSK), glycoprotein IIb/IIIa, glycoprotein Ib/IX, red blood cell antigens, platelet antigens and the like.

[0040] Methods for determining the level of autoantibodies are known in the art and any known method can be used. Examples of suitable methods include immunological methods such as enzyme-linked immunosorbent assays (ELISA), lateral flow immunoassays (LFIA, also known as lateral flow immunochromatographic assays), diffusion immunoassays (DIA), fluoroimmunoassays (FIA) , chemiluminescent immunoassays (CLIA), counting immunoassays (CIA), magnetic immunoassays (MIA), radioimmunoassays (RIA) and the like. For example, a detectably labeled autoantigen can be used in an assay to detect an autoantibody, respectively. The autoantibody present in a biological sample obtained from an individual being treated can be immobilized and the detectably labeled autoantigen can be brought into contact with the immobilized autoantibody forming a complex, where the presence or amount of detectable marker indicates the presence or amount of autoantibody in the biological sample.

[0041] In some cases, a treatment method of the present invention comprises: a) administering to the individual an antibody that specifically binds to complement C1s in an amount and for a period effective to reduce the level of autoantibody titers; and b) detect a level of autoantibodies in a biological sample obtained from the individual. A level of autoantibodies in a biological sample obtained from the individual that is lower than a pretreatment level may indicate the effectiveness of the treatment. A level of autoantibodies in a biological sample obtained from the individual that is not significantly lower than a pretreatment level may indicate the need to increase the dose and/or duration of administration and/or frequency of administration. A level of autoantibodies in a biological sample obtained from the individual that is greater than the pretreatment level may indicate the need to increase the dose and/or duration of administration and/or frequency of administration.

[0042] In some cases, a treatment method of the present invention comprises: a) administering to the individual an antibody that specifically binds to complement C1s in an amount and for a period effective to reduce the level of autoantibody titers; b) detect a level of autoantibodies in a biological sample obtained from the individual; and c) adjust the dose of anti-C1s antibody based on the level detected.

[0043] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and over a period of time to an individual with an autoimmune disorder, is effective in reducing the level of activation of B cells in the individual by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50% , at least 60%, at least 70%, or at least 80% compared to the level of B cell activation in the subject in the absence of anti-C1s antibody treatment or compared to the level of B cell activation in the individual before treatment with the anti-C1s antibody.

[0044] The present invention provides a method of reducing B cell activation in an individual with an autoimmune disorder, the method comprising administering to the individual an effective amount of an antibody specific against the C1s component of complement. Anti-C1s antibody is administered in an amount and for a period of time effective to reduce the level of B cell activation. In some cases, the effectiveness of treatment is monitored after administration of the anti-C1s antibody. In some cases, the anti-C1s antibody dose is adjusted based on monitoring results. Thus, in some cases, a method of the present invention comprises: a) administering to the subject an effective amount of an antibody specific for the C1s component of complement; and b) monitoring the effectiveness of said administration comprising detecting a level of B cell activation in a biological sample obtained from the individual. In some cases, a method of the present invention comprises: a) administering to the subject an effective amount of an antibody specific for the C1s complement component; b) monitoring the effectiveness of said administration comprising detecting a level of B cell activation in a biological sample obtained from the individual; and c) adjusting the dose of the anti-C1s antibody based on the detected level of B cell activation. The biological sample comprises B cells. For example, the biological sample may be a blood sample or other liquid or tissue sample that contains B cells B cells can be isolated from the biological sample.

[0045] B cell activation can be determined using any convenient method, including, for example, calcium flow. Calcium flux can be determined using a fluorescent calcium indicator. Fluorescent calcium indicators are known in the art and include, but are not limited to, fura-2, bis-fura 2, indo-1, Quin-2, Quin-2 AM, Benzothiaza-1, Benzothiaza-2, indo-5F , Fura-FF, BTC, Mag-Fura-2, Mag-Fura-5, Mag-Indo-1, fluo-3, rhod-2, fura-4F, fura-5F, fura-6F, fluo-4, fluo -5F, fluo-5N, Oregon Green 488 BAPTA, Calcium Green, Calcein, Fura-C18, Calcium Green-C18, Calcium Orange, Calcium Crimson, Calcium Green-5N, Magnesium Green, Oregon Green 488 BAPTA -1, Oregon Green 488 BAPTA-2, X-rhod-1, Fura Red, Rhod-5F, Rhod-5N, X-Rhod-5N, Mag-Rhod-2, Mag- 5N, Fluo-5F, Fluo-4FF, Mag-Fluo-4, Aequorin, dextran conjugates or any other derivatives of any of these dyes and others (see, for example, the catalog or website of Molecular Probes, Eugene, see , also, Nuccitelli, ed., Methods in Cell Biology, Volume 40: A Practical Guide to the Study of Calcium in Living Cells, Academic Press (1994); , Humana Press (1999); W. T. Mason, ed., Fluorescent and Luminescent Probes for Biological Activity. A Practical Guide to Technology for Quantitative Real-Time Analysis, Second Ed, Academic Press (1999); Calcium Signaling Protocols (Methods in Molecular Biology), 2005, D. G. Lamber, ed., Humana Press.).

[0046] B cell activation can be determined using other convenient methods, including, for example, assessment of cell surface markers of B cell activation and differentiation. Cell surface activation markers include, but are not limited to, CD23, CD25, CD27, CD30, CD38, CD69, CD80, CD86, CD135 and the like, which can be monitored using flow cytometry, immunohistochemistry, immunofluorescence and other methods used in the field. Furthermore, cell surface markers that are specific for naïve and undifferentiated B cells can be monitored to assess the proportion of naïve versus activated cells in the circulation. Naive cell markers include, but are not limited to, IgM, CD10 and other such markers. Additionally, intracellular activation markers such as transcription factors, phosphosignaling proteins, and cytokines can also be monitored to assess the activation and proliferative state of B cells. Transcription factors that can be monitored include, but are not limited to, Oct-2, Pax-5, Blimp-1, Bcl-6, XPB-1 and the like. Phosphosignaling proteins that can be monitored include, but are not limited to, phospho-Akt, phospho-Btk, phospho-Syk, phospho - BLNK, phospho-CD20/BL-CAM, phospho-IKKY, phospho-NFKB, phospho-mTOR and the like. Cytokines that can be monitored include, but are not limited to, IL-2, IL-4, IL-6, IFN-Y, IL-10, IL-12, TNF-α, TGF-β and the like. The evaluation of transcription factors, phosphosignaling proteins and cytokines can be assessed using flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), cell immunofluorescence, as well as immunoabsorption assays. enzymatic analysis (ELISAs) of cytokine levels assessed in whole blood, plasma or serum of patients, and other methods that are known in the field. Additionally, the size and granularity of B cells can be monitored by flow cytometry, microscopy and other methods known in the field to assess the activation status of B cells.

[0047] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and for a period of time to an individual with an autoimmune disorder, is effective in reducing cell proliferation B in the individual by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least at least 60%, at least 70%, or at least 80% compared to the level of B cell proliferation in the subject in the absence of anti-C1s antibody treatment or compared to the level of B cell activation in the subject before treatment with anti-C1s antibody.

[0048] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and over a period of time to an individual with an autoimmune disorder, is effective in reducing the number of cells B autoreactive in the individual in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, or at least 80% compared to the number of autoreactive B cells in the subject in the absence of anti-C1s antibody treatment or compared to the number of autoreactive B cells in the subject before treatment with anti-C1s antibody.

[0049] The present invention provides a method of reducing the proliferation of B cells in an individual with an autoimmune disorder, the method comprising administering to the individual an effective amount of an antibody specific against the C1s component of complement. The anti-C1s antibody is administered in an amount and for a period of time effective to reduce the level of B cell proliferation. In some cases, the effectiveness of the treatment is monitored after administration of the anti-C1s antibody. In some cases, the anti-C1s antibody dose is adjusted based on monitoring results. Thus, in some cases, a method of the present invention comprises: a) administering to the subject an effective amount of an antibody specific for the C1s component of complement; and b) monitoring the effectiveness of said administration comprising detecting a level of B cell proliferation in a biological sample obtained from the individual. In some cases, a method of the present invention comprises: a) administering to the subject an effective amount of an antibody specific for the C1s complement component; b) monitoring the effectiveness of said administration comprising detecting a level of B cell proliferation in a biological sample obtained from the individual; and c) adjusting the dose of the anti-C1s antibody based on the detected level of B cell proliferation. The biological sample comprises B cells. For example, the biological sample may be a blood sample or other liquid or tissue sample that contains cells B. B cells can be isolated from the biological sample.

[0050] B cell proliferation can be determined using any known assay, for example, by determining the number of CD19+ or CD20+ or CD21+ B cells or CD22+ B cells (e.g., using flow cytometry, microscopy, fluorescent microscopy, a hemocytometer and other instruments and methods known in the field).

[0051] Autoimmune disorders that can be treated using a method of the present invention for treating an autoimmune disorder are antibody-mediated autoimmune disorders and include, but are not limited to, Addison's disease, age-related macular degeneration, alopecia , autoimmune hepatitis (e.g., autoimmune hepatitis associated with hepatitis B virus infection; autoimmune hepatitis associated with hepatitis C virus infection), autoimmune hemolytic anemia, autoimmune skin diseases, autoimmune thyroid disease, bullous pemphigus, celiac disease, cold agglutinin disease, dermatomyositis, type 1 diabetes mellitus, Grave's disease, Goodpasture syndrome, Hashimoto's disease, hypoparathyroidism, hypopituitarism, hypothyroidism, idiopathic thrombocytopenic purpura, inflammatory bowel disease (e.g., Crohn's disease, colitis ulcerative), multiple sclerosis, myasthenia gravis, myocarditis, neuromyelitis optica, pemphigus vulgaris, pemphigus foliaceus, polymyositis, psoriasis, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, uveitis and Wegener's granulomatosis and poly /dermatomyositis.

[0052] Diseases that can be treated using a method of the present invention include, for example, age-related autoimmune disorders, age-related macular degeneration, Alzheimer's disease, amyotrophic lateral sclerosis, anaphylaxis, argyrophilic grain dementia, arthritis (e.g. , rheumatoid arthritis), asthma, atherosclerosis, atypical hemolytic-uremic syndrome, autoimmune diseases, autoimmune hemolytic anemia, Barraquer-Simons syndrome, Behçet's disease, British-type amyloid angiopathy, bullous pemphigus, Buerger's disease, C1q nephropathy, cancer, catastrophic antiphospholipid syndrome, cerebral amyloid angiopathy, cold agglutinin disease, corticobasal degeneration, Creutzfeldt-Jakob disease, Crohn's disease, cryoglobulinemic vasculitis, dementia pugilistica, dementia with Lewy bodies (DLB), diffuse neurofibrillary tangles with calcification , discoid lupus erythematosus, Down syndrome, focal segmental glomerulosclerosis, formal thought disorder, frontotemporal dementia (FTD), frontotemporal dementia with parkinsonism linked to chromosome 17, frontotemporal lobar degeneration, Gerstmann-Straussler-Scheinker disease, Guillain syndrome -Barré, Hallervorden-Spatz disease, hemolytic uremic syndrome, hereditary angioedema, hypophosphatasis, idiopathic pneumonia syndrome, immune complex diseases, inclusion body myositis, infectious diseases (e.g., disease caused by bacteria (e.g. , Neisseria meningitidis or Streptococcus) virus (e.g., human immunodeficiency virus (HIV)), or other infectious agents), inflammatory disease, ischemia/reperfusion injury, mild cognitive impairment, immunothrombocytopenic purpura (ITP), molybdenum cofactor deficiency (MoCD) type A, membranoproliferative glomerulonephritis (MPGN) I, membranoproliferative glomerulonephritis (MPGN) II (dense deposit disease), membranous nephritis, multi-infarct dementia, lupus (e.g., systemic lupus erythematosus (SLE)), glomerulonephritis , Kawasaki disease, multifocal motor neuropathy, multiple sclerosis, multiple system atrophy, myasthenia gravis, myocardial infarction, myotonic dystrophy, neuromyelitis optica, Niemann-Pick disease type C, non-Guamanian motor neuron disease with neurofibrillary tangles, Parkinson's disease, Parkinson's disease with dementia, paroxysmal nocturnal hemoglobinuria, pemphigus vulgaris, Pick's disease, post-encephalitis parkinsonism, polymyositis, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, progressive supranuclear palsy, psoriasis, sepsis, Shiga toxin E. coli (STEC)-HuS, spinal muscular atrophy, subacute sclerosing panencephalitis, stroke, tangle-only dementia, transplant rejection, vasculitis (e.g., ANCA-associated vasculitis), Wegner's granulomatosis, sickle cell disease, cryo- globulinemia, mixed cryoglobulinemia, essential mixed cryoglobulinemia, mixed cryoglobulinemia type II, mixed cryoglobulinemia type III, nephritis, drug-induced thrombocytopenia, lupus nephritis, bullous pemphigus, epidermolysis bullosa acquisita, delayed hemolytic transfusion reaction, hypocomplementemic urticarial vasculitis syndrome , pseudophakic bullous keratopathy and platelet refractoriness.

Reducing the Level of Alloimmune Antibodies

[0053] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and over a period of time to an individual in need thereof (e.g., a transplant graft or recipient of an organ), is effective in reducing the level of alloantibodies in the individual by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40 %, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or more than 90% compared to the alloantibody level in the individual in the absence of treatment with the anti-C1s antibody or compared to the level of alloantibodies in the individual before treatment with the anti-C1s antibody.

[0054] A method of the present invention provides a reduction in the level of alloantibodies in an individual. Alloantibodies include antibodies against human leukocyte antigen (HLA) present in a donor tissue or organ. Alloantibodies include antibodies against any epitope present on a donor tissue, donor organ, or donor cell (e.g., red blood cells, platelets, endothelial cells, etc.).

[0055] Methods for determining the level of alloantibodies are known in the art and any known method can be used. Examples of suitable methods include immunological methods such as ELISA, LFIA, DIA, FIA, CLIA, CIA, MIA, RIA and the like. For example, a detectably labeled alloantigen can be used in an assay to detect an alloantibody, respectively. The alloantibody present in a biological sample obtained from an individual being treated can be immobilized and the detectably labeled alloantigen can be brought into contact with the immobilized alloantibody forming a complex, where the presence or amount of detectable marker indicates the presence or amount of alloantibody in the biological sample.

[0056] In some cases, a treatment method of the present invention comprises: a) administering to the individual an antibody that specifically binds to complement C1s in an amount and for a period effective to reduce the level of alloantibody titers; and b) detect a level of alloantibodies in a biological sample obtained from the individual. A level of alloantibodies in a biological sample obtained from the individual that is lower than a pretreatment level may indicate the effectiveness of the treatment. A level of alloantibodies in a biological sample obtained from the subject that is not significantly less than a pretreatment level may indicate the need to increase the dose and/or duration of administration and/or frequency of administration. A level of alloantibodies in a biological sample obtained from the individual that is higher than the pre-treatment level may indicate the need to increase the dose and/or duration of administration and/or frequency of administration.

[0057] In some cases, a treatment method of the present invention comprises: a) administering to the individual an antibody that specifically binds to complement C1s in an amount and for a period effective to reduce the level of alloantibody titers; b) detect a level of alloantibodies in a biological sample obtained from the individual; and c) adjust the dose of anti-C1s antibody based on the level detected.

[0058] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and over a period of time to an individual with an alloimmune disorder, is effective in reducing the level of activation of B cells in the individual by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, or at least 80% compared to the level of B cell activation in the subject in the absence of anti-C1s antibody treatment or compared to the level of cell activation B in the individual before treatment with the anti-C1s antibody.

[0059] The present invention provides a method of reducing B cell activation in an individual with an alloimmune disorder, the method comprising administering to the individual an effective amount of an antibody specific against the C1s component of complement. Anti-C1s antibody is administered in an amount and for a period of time effective to reduce the level of B cell activation. In some cases, the effectiveness of treatment is monitored after administration of the anti-C1s antibody. In some cases, the dose of anti-C1s antibody is adjusted based on monitoring results. Thus, in some cases, a method of the present invention comprises: a) administering to the subject an effective amount of an antibody specific for the C1s component of complement; and b) monitoring the effectiveness of said administration comprising detecting a level of B cell activation in a biological sample obtained from the individual. In some cases, a method of the present invention comprises: a) administering to the subject an effective amount of an antibody specific for the C1s complement component; b) monitoring the effectiveness of said administration comprising detecting a level of B cell activation in a biological sample obtained from the individual; and c) adjusting the dose of the anti-C1s antibody based on the detected level of B cell activation. The biological sample comprises B cells. For example, the biological sample may be a blood sample or other liquid or tissue sample that contains B cells B cells can be isolated from the biological sample.

[0060] B cell activation can be determined using any convenient method, including, for example, calcium flow. Calcium flux can be determined using a fluorescent calcium indicator. Fluorescent calcium indicators are known in the art and include, but are not limited to, fura-2, bis-fura 2, indo-1, Quin-2, Quin-2 AM, Benzothiaza-1, Benzothiaza-2, indo-5F , Fura-FF, BTC, Mag-Fura-2, Mag-Fura-5, Mag-Indo-1, fluo-3, rhod-2, fura-4F, fura-5F, fura-6F, fluo-4, fluo -5F, fluo-5N, Oregon Green 488 BAPTA, Calcium Green, Calcein, Fura-C18, Calcium Green-C18, Calcium Orange, Calcium Crimson, Calcium Green-5N, Magnesium Green, Oregon Green 488 BAPTA -1, Oregon Green 488 BAPTA-2, X-rhod-1, Fura Red, Rhod-5F, Rhod-5N, X-Rhod-5N, Mag-Rhod-2, Mag- 5N, Fluo-5F, Fluo-4FF, Mag-Fluo-4, Aequorin, dextran conjugates or any other derivatives of any of these dyes and others (see, for example, the catalog or website of Molecular Probes, Eugene , see also, Nuccitelli, ed., Methods in Cell Biology, Volume 40: A Practical Guide to the Study of Calcium in Living Cells, Academic Press (1994); 114), Humana Press (1999); W. T. Mason, ed., Fluorescent and Luminescent Probes for Biological Activity. A Practical Guide to Technology for Quantitative Real-Time Analysis, Second Ed, Academic Press (1999); Calcium Signaling Protocols (Methods in Molecular Biology), 2005, D. G. Lamber, ed., Humana Press.).

[0061] B cell activation can be determined using or other convenient methods, including, for example, assessment of cell surface markers of B cell activation and differentiation. Cell surface activation markers include, but are not limited to , CD23, CD25, CD27, CD30, CD38, CD69, CD80, CD86, CD135 and the like, which can be monitored using flow cytometry, immunohistochemistry, immunofluorescence and other methods used in the field. Furthermore, cell surface markers that are specific for naïve and undifferentiated B cells can be monitored to assess the proportion of naïve versus activated cells in the circulation. Naive cell markers include, but are not limited to, IgM, CD10 and other such markers. Additionally, intracellular activation markers such as transcription factors, phosphosignaling proteins, and cytokines can also be monitored to assess the activation and proliferative state of B cells. Transcription factors that can be monitored include, but are not limited to, Oct-2, Pax-5, Blimp-1, Bcl-6, XPB-1 and the like. Phosphosignaling proteins that can be monitored include, but are not limited to, phospho-Akt, phospho-Btk, phospho-Syk, phospho - BLNK, phospho-CD20/BL-CAM, phospho-IKKY, phospho-NFKB, phospho-mTOR and the like. Cytokines that can be monitored include, but are not limited to, IL-2, IL-4, IL-6, IFN-, IL-10, IL-12, TNF-, TGF- and the like. Assessment of transcription factors, phosphosignaling proteins and cytokines can be assessed using flow cytometry, RT-PCR, cell immunofluorescence, as well as ELISAs of cytokine levels assessed in whole blood, plasma or serum of patients and other methods known in the field. Additionally, the size and granularity of B cells can be monitored by flow cytometry, microscopy, and other methods known in the field to assess the activation status of B cells.

[0062] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and over a period of time to an individual with an alloimmune disorder, is effective in reducing proliferation of B cells in the individual by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50% , at least 60%, at least 70%, or at least 80% compared to the level of B cell proliferation in the subject in the absence of anti-C1s antibody treatment or compared to the level of B cell activation in the subject individual before treatment with the anti-C1s antibody.

[0063] In some cases, an effective amount of an anti-C1s antibody is an amount that, when administered in one or more doses and over a period of time to an individual with an alloimmune disorder, is effective in reducing the number of alloreactive B cells in the individual by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least at least 50%, at least 60%, at least 70%, or at least 80% compared to the number of alloreactive B cells in the subject in the absence of anti-C1s antibody treatment or compared to the number of alloreactive B cells in the individual before treatment with the anti-C1s antibody.

[0064] The present invention provides a method of reducing the proliferation of B cells in an individual with an alloimmune disorder, the method comprising administering to the individual an effective amount of an antibody specific against the C1s component of complement. The anti-C1s antibody is administered in an amount and for a period of time effective to reduce the level of B cell proliferation.

[0065] B cell proliferation can be determined using any known assay, for example, by determining the number of CD19+ or CD20+ or CD21+ B cells or CD22+ B cells (e.g., using flow cytometry, microscopy, fluorescent microscopy, a hemocytometer and other instruments and methods known in the field).

[0066] Alloimmune disorders that can be treated using a method of the present invention for treating an alloimmune disorder include antibody-mediated rejection of an allografted organ, tissue or cell. Allografted organs, tissues, and cells include, but are not limited to, a kidney, a liver, a pancreas, a heart, a lung, skin, blood tissue (including whole blood, red blood cells, white blood cells, umbilical cord blood, and similar), where the blood tissue may comprise an isolated population of blood cells (buffy coat, red blood cells, platelets, lymphocytes, T cells, B cells or some other population), or where the blood tissue comprises a mixed population of cells), small intestine, an endothelial tissue, a vascular tissue (e.g., a blood vessel), an eye, a stomach, a thymus, bone, bone marrow, cornea, a heart valve, an islet of Langerhans, or a tendon. As used herein, "organ" encompasses an entire organ or a part of an organ. As used herein, "fabric" encompasses an entire fabric or part of a fabric.

[0067] In some cases, a method of the present invention for treating an alloimmune disorder comprises administering an effective amount of an anti-C1s antibody to an individual who has received an organ or tissue from a donor (e.g., a recipient of an organ or fabric). In some cases, a method of the present invention for treating an alloimmune disorder comprises administering an effective amount of an anti-C1s antibody to an individual who has received a donor organ or tissue (e.g., a recipient of an organ or tissue), where the individual exhibits symptoms of antibody-mediated rejection (AMR). In some cases, a method of the present invention for treating an alloimmune disorder comprises administering an effective amount of an anti-C1s antibody to an individual who has received a donor organ or tissue (e.g., a recipient of an organ or tissue), where the individual has been diagnosed with AMR. Thus, for example, in some cases, the present invention provides a method of treating AMR, comprising administering to an individual who has been diagnosed as having AMR an effective amount of an anti-C1s antibody. In some cases, a method of the present invention provides a reduction in B cell proliferation and/or B cell activation in an individual with AMR.

[0068] In some cases, a method of the present invention for treating an alloimmune disorder comprises administering an effective amount of an anti-C1s antibody to an individual who is to receive (e.g., who is scheduled to receive; who is on a list waiting to receive; etc.) a donor organ, a donor tissue, or a donor cell (or population of donor cells) (e.g., a prospective recipient of an organ or tissue; a prospective recipient of a transfusion; a prospective bone marrow transplant recipient, etc.). In some cases, a method of the present invention for treating an alloimmune disorder comprises administering an effective amount of an anti-C1s antibody to an individual who is to receive (e.g., who is scheduled to receive; who is on a list of waiting to receive; etc.) a donor organ, a donor tissue, or a donor cell (or population of donor cells) (e.g., a prospective recipient of an organ or tissue; a prospective recipient of bone marrow transplantation; etc.), where treatment with the anti-C1s antibody begins before the individual receives the donor organ, donor tissue, or donor cell or cell population, and where treatment continues afterward. individual has received the donor organ, donor tissue, or donor cell or cell population.

[0069] In some cases, when carrying out a method of the present invention to treat an alloimmune disorder, an anti-C1s antibody is administered to a prospective recipient organ or tissue from 1 hour to 7 days (e.g., from 1 hour to 4 hours, from 4 hours to 8 hours, from 8 hours to 12 hours, from 12 hours to 16 hours, from 16 hours to 24 hours, from 1 day to 2 days, from 2 days to 3 days, from 3 days to 4 days, from 4 days to 5 days, from 5 days to 6 days, or from 6 days to 7 days) before receiving the organ or tissue.

Dosages, frequency of administration; duration of administration

[0070] A suitable dosage of an anti-C1s antibody can be determined by a physician or other qualified medical personnel, based on various clinical factors. As is well known in the medical arts, dosages for any patient depend on many factors, including size, body surface area, age of the patient, the specific compound to be administered, sex of the patient, time and route of administration, general health, and other drugs being administered concomitantly. An anti-C1s antibody can be administered in amounts between 1 ng/kg of body weight and 100 mg/kg of body weight per dose, for example, from 1 ng/kg of body weight to 50 ng/kg of body weight, of 50 ng/kg body weight to 0.1 mg/kg body weight, from 0.1 mg/kg body weight to 1 mg/kg body weight, from 1 mg/kg body weight to 5 mg/kg of body weight, from 5 mg/kg of body weight to 10 mg/kg of body weight, from 0.5 mg/kg of body weight to 5 mg/kg of body weight, from 10 mg/kg of body weight to 20 mg/kg body weight, from 20 mg/kg body weight to 50 mg/kg body weight, or from 50 mg/kg body weight to 100 mg/kg body weight, or more than 100 mg/kg of body weight; however, doses below or above this sample range are anticipated, especially considering the aforementioned factors. If the regimen is a continuous infusion, it may also be in the range of 1 μg to 10 mg per kilogram of body weight per minute.

[0071] In some cases, a dose of an anti-C1s antibody is in the range of 0.001 μg to 1000 μg; however, doses below or above this example range are expected, especially considering the factors mentioned above. In some cases, the dosage may vary, for example, from about 0.0001 to 100 mg/kg, or from 0.01 to 5 mg/kg (e.g., 0.02 mg/kg, 0.25 mg/kg). kg, 0.5 mg/kg, 0.75 mg/kg, 1 mg/kg, 2 mg/kg, etc.) of body weight. For example, dosages may be 1 mg/kg of body weight or 10 mg/kg of body weight or within the range of 1-10 mg/kg or at least 1 mg/kg. It is also intended that intermediate doses in the above ranges are considered to be within the scope of the invention.

[0072] In some embodiments, an anti-C1s antibody is administered in an amount that provides a peak serum concentration of about 1 μg/mL to about 1 mg/mL, for example, of about 1 μg/mL to about 2.5 μg/mL, from about 2.5 μg/mL to about 5 μg/mL, from about 5 μg/mL to about 7.5 μg/mL, from about 7.5 μg/mL to about 10 μg/mL, from about 10 μg/mL to about 25 μg/mL, from about 25 μg/mL to about 50 μg/mL, from about 50 μg/mL to about from 100 μg/mL, from about 100 μg/mL to about 250 μg/mL, from about 250 μg/mL to about 500 μg/mL, from about 500 μg/mL to about 750 μg/mL or from about 750 μg/mL to about 1000 μg/mL. In some embodiments, an anti-C1s antibody is administered in an amount that provides a maximum serum concentration greater than 1 mg/mL, e.g., from about 1 mg/mL to about 2 mg/mL, from about 2 mg /mL to about 5 mg/mL, or from about 5 mg/mL to about 10 mg/mL.

[0073] An anti-C1s antibody formulation can be administered at a variety of frequencies. In some cases, multiple doses of an anti-C1s antibody are administered. The frequency of administration of an anti-C1s antibody may vary depending on any of a variety of factors, for example, the severity of symptoms, etc. For example, in some embodiments, an anti-C1s antibody is administered once a month, twice a month, three times a month, every other week (qow), once a week (qw), twice a week (biw), three times a week (tiw), four times a week, five times a week, six times a week, every other day (qod), daily (qd), twice a day (qid), or three times per day (tid).

[0074] In some cases, an anti-C1s antibody is administered for a period of 6 months or more. In some cases, an anti-C1s antibody is administered for a period of 6 months to 1 year, 1 year to 2 years, 2 years to 5 years, or longer than 5 years.

[0075] In some cases, an anti-C1s antibody is administered for a period of less than 6 months. In some cases, an anti-C1s antibody is administered for a period of 5.5 months or less. In some cases, an anti-C1s antibody is administered for a period of 5 months or less. In some cases, an anti-C1s antibody is administered for a period of 4.5 months or less. In some cases, an anti-C1s antibody is administered for a period of 4 months or less. In some cases, an anti-C1s antibody is administered for a period of 3.5 months or less. In some cases, an anti-C1s antibody is administered for a period of 3 months or less. In some cases, an anti-C1s antibody is administered for a period of 2.5 months or less. In some cases, an anti-C1s antibody is administered for a period of 2 months or less. In some cases, an anti-C1s antibody is administered for a period of 1 month or less. In some cases, an anti-C1s antibody is administered for a period of 3 weeks. In some cases, an anti-C1s antibody is administered for a period of 2 weeks. In some cases, an anti-C1s antibody is administered for a period of 1 week.

[0076] An anti-C1s antibody formulation can be administered through a variety of administration routes. Conventional and pharmaceutically acceptable routes of administration include intranasal, intramuscular, intratracheal, intrathecal, intracranial, subcutaneous, intradermal, topical, intravenous, intraperitoneal, intra-arterial (e.g., via the carotid artery), spinal or cerebral delivery, rectal, nasal, oral and other enteral and parenteral routes of administration. Routes of administration can be combined, if desired, or adjusted depending on the antibody and/or desired effect. In some cases, an anti-C1s antibody is administered subcutaneously. In some cases, an anti-C1s antibody is administered intravenously. In some cases, an anti-C1s antibody is administered intramuscularly.

Anti-C1s antibodies

[0077] Any of a variety of anti-C1s antibodies can be used in a method of the present invention for treating an alloimmune disorder or an autoimmune disorder, or in a method of reducing B cell proliferation and/or B cell activation In some cases, the anti-C1s antibody is humanized. In some cases, the anti-C1s antibody comprises a humanized VH framework region. In some cases, the anti-C1s antibody comprises a humanized VL framework region. In some cases, the anti-C1s antibody comprises a humanized VH framework region and a humanized VL framework region.

[0078] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: VSA (SEQ ID NO:100).

[0079] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVLTQSTDYLAVSLGQRATISCKASQSVDYDGDSYMNWYQQKPGQ PPKLLIYAASNLESGIPARFSGSGSGTDFTLNIHPVEEEDAATYYCQQ SNEDPWTFGGGTKLEIK (SEQ ID NO: 101).

[0080] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[0081] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:1);

[0082] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:2); It is

[0083] 3) CDR-H3: AIYGSGWAWFPY (SEQ ID NO:3).

[0084] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[0085] 1) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:4);

[0086] 2) CDR-L2: AASNLESGIP (SEQ ID NO:5); It is

[0087] 3) CDR L3: QQSNEDPWT (SEQ ID NO:6).

[0088] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[0089] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:1);

[0090] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:2);

[0091] 3) CDR-H3: AIYGSGWAWFPY (SEQ ID NO:3);

[0092] 4) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:4);

[0093] 5) CDR-L2: AASNLESGIP (SEQ ID NO:5); It is

[0094] 6) CDR-L3: QQSNEDPWT (SEQ ID NO: 6).

[0095] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: TVSS (SEQ ID NO:7).

[0096] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: ID NO:8).

[0097] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[0098] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:9);

[0099] 2) CDR-H2: IDPADGHTKY (SEQ ID NO:10); It is

[00100] 3) CDR-H3: ARYGYGREVFDY (SEQ ID NO:11).

[00101] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00102] 1) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:12);

[00103] 2) CDR-L2: DASNLESGIP (SEQ ID NO:13); It is

[00104] 3) CDR-L3: QQSNEDPWT (SEQ ID NO:14).

[00105] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00106] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:9);

[00107] 2) CDR-H2: IDPADGHTKY (SEQ ID NO:10);

[00108] 3) CDR-H3: ARYGYGREVFDY (SEQ ID NO:11);

[00109] 4) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:12);

[00110] 5) CDR-L2: DASNLESGIP (SEQ ID NO:13); It is

[00111] 6) CDR-L3: QQSNEDPWT (SEQ ID NO:14).

[00112] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: YGWFAYWGQGTLVTVSA (SEQ ID NO: 102).

[00113] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVMTQSPAIMSASPGERVTMTCSASSSISYMHWYHQKPGTSPKRWI YDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCHQRSSFP TFGAGTKLELK (SEQ ID NO:103).

[00114] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00115] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:15);

[00116] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:16); It is

[00117] 3) CDR-H3: TIDDSAYGWFAY (SEQ ID NO:17).

[00118] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00119] 1) CDR-L1: SSISYMHWYHQK (SEQ ID NO:18);

[00120] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:19); It is

[00121] 3) CDR-L3: HQRSSFPT (SEQ ID NO:20).

[00122] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00123] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:15;

[00124] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:16);

[00125] 3) CDR-H3: TIDDSAYGWFAY (SEQ ID NO:17).

[00126] 4) CDR-L1: SSISYMHWYHQK (SEQ ID NO:18);

[00127] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:19); It is

[00128] 6) CDR-L3: HQRSSFPT (SEQ ID NO:20).

[00129] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: YWGQGTLVTVSA (SEQ ID NO:104) .

[00130] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVITQSPAIMSAS- PGERVTMTCSASSSISYMHWYHQKPGTSPKRWIYDTSKLASGV- PARFSGSGGSTSYSLTISSMEAEDAATYYCHQRSSFPTFGAGT- KLELK (SEQ ID NO:105) .

[00131] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00132] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:21);

[00133] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:22); It is

[00134] 3) CDR-H3: TIDDSVYGWFAY (SEQ ID NO:23).

[00135] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00136] 1) CDR-L1: SSISYMHWYHQK (SEQ ID NO:24);

[00137] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:25); It is

[00138] 3) CDR-L3: HQRSSFPT (SEQ ID NO:26).

[00139] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00140] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:21);

[00141] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:22);

[00142] 3) CDR-H3: TIDDSVYGWFAY (SEQ ID NO:23);

[00143] 4) CDR-L1: SSISYMHWYHQK (SEQ ID NO:24);

[00144] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:25); It is

[00145] 6) CDR-L3: HQRSSFPT (SEQ ID NO:26).

[00146] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: FPYWGQGTLVSVSAAKTTAP- SVYPLAPVCGDTTGSSVTLGCLVK (SEQ ID NO: 106).

[00147] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: SEQ ID NO:107) .

[00148] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00149] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:27);

[00150] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:28); It is

[00151] 3) CDR-H2: AIYGSGWAWFPY (SEQ ID NO:29).

[00152] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00153] 1) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:30);

[00154] 2) CDR-L2: AASNLEFGIP (SEQ ID NO:31); It is

[00155] 3) CDR-L3: QQSNEDPWT (SEQ ID NO:32).

[00156] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00157] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:27);

[00158] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:28);

[00159] 3) CDR-H2: AIYGSGWAWFPY (SEQ ID NO:29);

[00160] 4) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:30);

[00161] 5) CDR-L2: AASNLEFGIP (SEQ ID NO:31); It is

[00162] 6) CDR-L3: QQSNEDPWT (SEQ ID NO:32).

[00163] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: DYWGQGTTLTVSS (SEQ ID NO:108) .

[00164] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVLTQFPT- FLAVFLGQRAPISCKASQSVDYDGDSYMNWFQQKTGQPPKI- LIYDASNLEFGIPTRFSGSGFGTDFPLNIHPVEEEDAAIYFCQQSNED- PWTFGGGPKLEIK (SEQ ID NO :109) .

[00165] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00166] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:33);

[00167] 2) CDR-H2: IDPADGHTKY (SEQ ID NO:34); It is

[00168] 3) CDR-H3: ARYGYGREVFDY (SEQ ID NO:35).

[00169] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00170] 1) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:36);

[00171] 2) CDR-L2: DASNLEFGIP (SEQ ID NO:37); It is

[00172] 3) CDR-L3: QQSNEDPWT (SEQ ID NO:38).

[00173] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00174] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:33);

[00175] 2) CDR-H2: IDPADGHTKY (SEQ ID NO:34);

[00176] 3) CDR-H3: ARYGYGREVFDY (SEQ ID NO:35);

[00177] 4) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:36);

[00178] 5) CDR-L2: DASNLEFGIP (SEQ ID NO:37); It is

[00179] 6) CDR-L3: QQSNEDPWT (SEQ ID NO:38).

[00180] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: WGQGTLVSVSA (SEQ ID NO:110) .

[00181] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: EFAMT- QSTDYLAVSLGQRATISCKASQSVDYDGDSYMNWYQQK- PGQPPKLLIYAASNLESGIPTRFSGSGFGTDFTLNIHPVEEEDAA- TYYCQQSNEDPWTFGGGPKLEI K (SEQ ID NO:111) .

[00182] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00183] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:39);

[00184] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:40); It is

[00185] 3) AIYGSGWAWFPY (SEQ ID NO:41).

[00186] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00187] 1) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:42);

[00188] 2) CDR-L2: AASNLESGIP (SEQ ID NO:43); It is

[00189] 3) CDR-L3: QQSNEDPWT (SEQ ID NO:44).

[00190] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00191] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:39);

[00192] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:40);

[00193] 3) AIYGSGWAWFPY (SEQ ID NO:41);

[00194] 4) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:42);

[00195] 5) CDR-L2: AASNLESGIP (SEQ ID NO:43); It is

[00196] 6) CDR-L3: QQSNEDPWT (SEQ ID NO:44).

[00197] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: WGQGTLVTVSA (SEQ ID NO:112) .

[00198] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVLTQT- TAIMSASPGEKVTMTCSASSSISYMYWFQQKPGTSPKR- WIYDTSKLASGVPARFSGSGSGTSYSLTISTMEAEDAA- TYYCHQRSSDPTFGGGTKLEINR (SEQ ID NO:113 ) .

[00199] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00200] 1) CDR-H1: GYSFTGYYIHWV (SEQ ID NO:45);

[00201] 2) CDR-H2: INPTTNDTTY (SEQ ID NO:46); It is

[00202] 3) CDR-H3: SRDISGPAWFAY (SEQ ID NO:47).

[00203] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00204] 1) CDR-L1: SSISYMYWFQQK (SEQ ID NO:48);

[00205] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:49);

[00206] 3) CDR-L3: HQRSSDPT (SEQ ID NO:50).

[00207] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00208] 1) CDR-H1: GYSFTGYYIHWV (SEQ ID NO:45);

[00209] 2) CDR-H2: INPTTNDTTY (SEQ ID NO:46);

[00210] 3) CDR-H3: SRDISGPAWFAY (SEQ ID NO:47);

[00211] 4) CDR-L1: SSISYMYWFQQK (SEQ ID NO:48);

[00212] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:49); It is

[00213] 6) CDR-L3: HQRSSDPT (SEQ ID NO:50).

[00214] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: GTLVTVSA (SEQ ID NO:114).

[00215] As an example of an appropriate anti-C1S antibody, in some cases the anti-C1S antibody comprises VL CDRs present in the following sequence of VL amino acids: DIVMTQSPaim- FASPGERVTMTCSSYPWYPQKPGPSPKRWYDTSKLAS QRSSFPPFGAGT- KLELK (SEQ ID NO: 115) .

[00216] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00217] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:51);

[00218] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:52); It is

[00219] 3) CDR-H3: TIDDSVYGWFAY (SEQ ID NO:53).

[00220] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00221] 1) CDR-L1: SSISY (SEQ ID NO:54);

[00222] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:55); It is

[00223] 3) CDR-L3: HQRSSFPP (SEQ ID NO:56).

[00224] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00225] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:51);

[00226] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:52);

[00227] 3) CDR-H3: TIDDSVYGWFAY (SEQ ID NO:53);

[00228] 4) CDR-L1: SSISY (SEQ ID NO:54);

[00229] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:55); It is

[00230] 6) CDR-L3: HQRSSFPP (SEQ ID NO:56).

[00231] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: AMDYWGQGTSVTVSS (SEQ ID NO:116) .

[00232] As an example of an appropriate anti-C1S antibody, in some cases the anti-C1S antibody comprises VL CDRs present in the following sequence of amino acids of VL: DIVMTQSPAIMSASPGEKVTMTCSSSYMHWYQQKPGSPGSPKSGSGSGSGSGSGSGSGSGSSMEAEDEDA QRSFYL TFGSGTKLIK (SEQ ID NO: 117).

[00233] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00234] 1) CDR-H1: GYTFTDYAMHCV (SEQ ID NO:57);

[00235] 2) CDR-H2: ISIYNGDASY (SEQ ID NO:58); It is

[00236] 3) CDR-H3: VREAPYLITTVFYAMDY (SEQ ID NO:59).

[00237] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00238] 1) CDR-L1: SSISYMHWYQQK (SEQ ID NO:60);

[00239] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:61); It is

[00240] 3) CDR-L3: HQRSFYLT (SEQ ID NO:62).

[00241] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00242] 1) CDR-H1: GYTFTDYAMHCV (SEQ ID NO:57);

[00243] 2) CDR-H2: ISIYNGDASY (SEQ ID NO:58);

[00244] 3) CDR-H3: VREAPYLITTVFYAMDY (SEQ ID NO:59);

[00245] 4) CDR-L1: SSISYMHWYQQK (SEQ ID NO:60);

[00246] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:61); It is

[00247] 6) CDR-L3: HQRSFYLT (SEQ ID NO:62).

[00248] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: GTLVTVSA (SEQ ID NO:118).

[00249] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVLTQSTAIMSASPGERVTMTCSASSSISYMHWYHQKPGTSPKRWI YDTSKLASGVPARFSGSGSGTSYSLAISSMEAEDAATYYCHQRSSFP TFGAGTKLELK (SEQ ID NO:119).

[00250] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00251] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:63);

[00252] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:64); It is

[00253] 3) CDR-H3: TIDDSAYGWFAY (SEQ ID NO:65).

[00254] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00255] 1) CDR-L1: SSISYMHWYHQK (SEQ ID NO:66);

[00256] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:67); It is

[00257] 3) CDR-L3: HQRSSFPT (SEQ ID NO:68).

[00258] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00259] 1) CDR-H1: GYTFTRYWMHWV (SEQ ID NO:63);

[00260] 2) CDR-H2: INPSNSDTDY (SEQ ID NO:64);

[00261] 3) CDR-H3: TIDDSAYGWFAY (SEQ ID NO:65);

[00262] 4) CDR-L1: SSISYMHWYHQK (SEQ ID NO:66);

[00263] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:67); It is

[00264] 6) CDR-L3: HQRSSFPT (SEQ ID NO:68).

[00265] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: SVSA (SEQ ID NO:120).

[00266] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVLTQTPDYLAVSLGQRATISCKASQSVDYDGDSYMNWYQQKPGQ PPKLLIYAASNLESGIPARFSGSGSGTDFTLNIHPVEEEDAATYYCQQ SNEDPWTFGGGTKLEIK (SEQ ID NO :121).

[00267] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00268] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:69);

[00269] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:70); It is

[00270] 3) CDR-H3: AIYGSGWAWFPY (SEQ ID NO:71).

[00271] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00272] 1) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:72);

[00273] 2) CDR-L2: AASNLESGIP (SEQ ID NO:73); It is

[00274] 3) CDR-L3: QQSNEDPWT (SEQ ID NO:74).

[00275] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00276] 1) CDR-H1: GFNIKDDYIHWV (SEQ ID NO:69);

[00277] 2) CDR-H2: IDPADDHTKY (SEQ ID NO:70);

[00278] 3) CDR-H3: AIYGSGWAWFPY (SEQ ID NO:71);

[00279] 4) CDR-L1: QSVDYDGDSYMN (SEQ ID NO:72);

[00280] 5) CDR-L2: AASNLESGIP (SEQ ID NO:73); It is

[00281] 6) CDR-L3: QQSNEDPWT (SEQ ID NO:74).

[00282] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: WGKDYLTVSA (SEQ ID NO:122) .

[00283] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVLT- QSPVIMSASPGEKVTMTCSASSSISYIHWYQQKPGTSPKR- WIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAA- TYYCHQRSSYLTFGSGTKLEIK (SEQ ID NO:12 3) .

[00284] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00285] 1) CDR-H1: GYSFTGFYMQWV (SEQ ID NO:75);

[00286] 2) CDR-H2: INPTTGDETY (SEQ ID NO:76); It is

[00287] 3) CDR-H3: ASDFYDGSFAWFEY (SEQ ID NO:77).

[00288] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00289] 1) CDR-L1: SSISYIHWYQQK (SEQ ID NO:78);

[00290] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:79); It is

[00291] 3) CDR-L3: HQRSSYLT (SEQ ID NO:80).

[00292] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00293] 1) CDR-H1: GYSFTGFYMQWV (SEQ ID NO:75);

[00294] 2) CDR-H2: INPTTGDETY (SEQ ID NO:76);

[00295] 3) CDR-H3: ASDFYDGSFAWFEY (SEQ ID NO:77);

[00296] 4) CDR-L1: SSISYIHWYQQK (SEQ ID NO:78);

[00297] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:79); It is

[00298] 6) CDR-L3: HQRSSYLT (SEQ ID NO:80).

[00299] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: FAYWGQGTLVTVSA (SEQ ID NO:124) .

[00300] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence: DIVMTQSPAIMSASSGEKVTMTCSASSSINYMHWYQQKPGTSPKRWI YDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDTATYYCHQRSDSL TFGSGTKLEIK (SEQ ID NO:125).

[00301] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00302] 1) CDR-H1: GYSFTGYYMHWV (SEQ ID NO:81);

[00303] 2) CDR-H2: INPSIGDITY (SEQ ID NO:82); It is

[00304] 3) CDR-H3: ASDYYGGGFAWFAY (SEQ ID NO:83).

[00305] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00306] 1) CDR-L1: SSINYMHWYQQK (SEQ ID NO:84);

[00307] 2) CDR-L2: DTSKLASGVP (SEQ ID NO:85); It is

[00308] 3) CDR-L3: HQRSDSLT (SEQ ID NO:86).

[00309] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00310] 1) CDR-H1: GYSFTGYYMHWV (SEQ ID NO:81);

[00311] 2) CDR-H2: INPSIGDITY (SEQ ID NO:82);

[00312] 3) CDR-H3: ASDYYGGGFAWFAY (SEQ ID NO:83);

[00313] 4) CDR-L1: SSINYMHWYQQK (SEQ ID NO:84);

[00314] 5) CDR-L2: DTSKLASGVP (SEQ ID NO:85); It is

[00315] 6) CDR-L3: HQRSDSLT (SEQ ID NO:86).

[00316] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00317] 1) CDR-H1: GFTFSNYAMSWV (SEQ ID NO:87);

[00318] 2) CDR-H2: ISSGGSHTYY (SEQ ID NO:88); It is

[00319] 3) CDR-H3: ARLFTGYAMDY (SEQ ID NO:89).

[00320] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00321] 1) CDR-L1: SSVSSSYLHWYQ (SEQ ID NO:90);

[00322] 2) CDR-L2: STSNLASGVP (SEQ ID NO:91); It is

[00323] 3) CDR-L3: HQYYRLPPIT (SEQ ID NO:92).

[00324] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00325] 1) CDR-H1: GFTFSNYAMSWV (SEQ ID NO:87);

[00326] 2) CDR-H2: ISSGGSHTYY (SEQ ID NO:88);

[00327] 3) CDR-H3: ARLFTGYAMDY (SEQ ID NO:89);

[00328] 4) CDR-L1: SSVSSSYLHWYQ (SEQ ID NO:90);

[00329] 5) CDR-L2: STSNLASGVP (SEQ ID NO:91); It is

[00330] 6) CDR-L3: HQYYRLPPIT (SEQ ID NO:92).

[00331] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VH CDRs present in the following VH amino acid sequence: EVMLVESGGALVKPGGSLKLSCAASGFTFSNYAMSWVRQIPEKRLE WVATISSGGSHTYYLDSVKGRFTISRDNARDTLYLQMSSLRSEDTAL YYCARLFTGYAMDYWGQGTSVTVSS (SEQ ID NO:93 )

[00332] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises VL CDRs present in the following VL amino acid sequence:

[00333] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs:

[00334] 1) CDR-H1: NYAMS (SEQ ID NO:95);

[00335] 2) CDR-H2: TISSGGSHTYYLDSVKG (SEQ ID NO:96); It is

[00336] 3) CDR-H3: LFTGYAMDY (SEQ ID NO:97).

[00337] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VL CDRs:

[00338] 1) CDR-L1: TASSSVSSSYLH (SEQ ID NO:98);

[00339] 2) CDR-L2: STSNLAS (SEQ ID NO:99); It is

[00340] 3) CDR-L3: HQYYRLPPIT (SEQ ID NO:92).

[00341] As an example of a suitable anti-C1s antibody, in some cases, the anti-C1s antibody comprises the following VH CDRs and VL CDRs:

[00342] 1) CDR-H1: NYAMS (SEQ ID NO:95);

[00343] 2) CDR-H2: TISSGGSHTYYLDSVKG (SEQ ID NO:96);

[00344] 3) CDR-H3: LFTGYAMDY (SEQ ID NO:97);

[00345] 4) CDR-L1: TASSSVSSSYLH (SEQ ID NO:98);

[00346] 5) CDR-L2: STSNLAS (SEQ ID NO:99); It is

[00347] 6) CDR-L3: HQYYRLPPIT (SEQ ID NO:92).

[00348] As noted above, in some cases, the anti-C1s antibody comprises a humanized VH framework region. In some cases, the anti-C1s antibody comprises a humanized VL framework region. In some cases, the anti-C1s antibody comprises a humanized VH framework region and a humanized VL framework region. Humanized VH and VL framework regions are known in the art and can be readily generated by those skilled in the art. In some cases, a humanized VH framework region is a consensus VH framework region. In some cases, a humanized VL framework region is a consensus VL framework region.

[00349] Non-limiting examples of human consensus VH framework regions suitable for use with VH CDRs as described herein include (subgroup III consensus):

[00350] a) FR1 VH: EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO:126);

[00351] b) FR2 VH: WVRQAPGKGLEWV (SEQ ID NO:127);

[00352] c) FR3 VH: RFTISRDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO:128); It is

[00353] d) FR4 VH: WGQGTLVTVSS (SEQ ID NO:129).

[00354] In some cases, FR3 VH comprises an amino acid substitution at position 71, 73 and/or 78; for example, where the bold underlined R in RFTISRDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO:130) is amino acid 71 (Kabat numbering); the underlined and bold N in RFTISRDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO:131) is amino acid 73 (Kabat numbering); and the underlined and bold L in RFTISRDNSKNTLYLQMNSLRAEDTAVYYC (SEQ ID NO:132) is amino acid 78 (Kabat numbering). For example, in some cases, amino acid 71 is A; and/or amino acid 73 is T; and/or amino acid 78 is A. As an example, in some cases, a suitable humanized consensus FR3 VH comprises the amino acid sequence: RFTISADTSKNTAYLQMNSLRAEDTAVYYC (SEQ ID NO:133).

[00355] Non-limiting examples of human consensus VH framework regions suitable for use with VH CDRs as described herein include (subgroup I consensus):

[00356] a) FR1 VH: QVQLVQSGAEVKKPGASVKVSCKAS (SEQ ID NO:134);

[00357] b) FR2 VH: WVRQAPGQGLEWM (SEQ ID NO:135);

[00358] c) FR3 VH: RVTITADTSTSTAYMELSSLRSEDTAVYYC (SEQ ID NO:136); It is

[00359] d) FR4 VH: WGQGTLVTVSS (SEQ ID NO:137).

[00360] Non-limiting examples of human consensus VH framework regions suitable for use with VH CDRs as described herein include (subgroup II consensus):

[00361] a) FR1 VH: QVQLQESGPGLVKPSQTLSLTCTVS (SEQ ID NO:138);

[00362] b) FR2 VH: WIRQPPGKGLEWI (SEQ ID NO:139);

[00363] c) FR3 VH: RVTISVDTSKNQFSLKLSSSVTAADTAVYYC (SEQ ID NO:140); It is

[00364] d) FR4 VH: WGQGTLVTVSS (SEQ ID NO:141).

[00365] Non-limiting examples of human consensus VL framework regions suitable for use with VL CDRs as described herein include (subgroup I consensus):

[00366] a) FR1 VL: DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO:142);

[00367] b) FR2 VL: WYQQKPGKAPKLLIY (SEQ ID NO:143);

[00368] c) FR3 VL: GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO:144); It is

[00369] d) FR4 VL: FGQGTKVEIK (SEQ ID NO:145).

[00370] Non-limiting examples of human consensus VL framework regions suitable for use with VL CDRs as described herein include (subgroup II consensus):

[00371] a) FR1 VL: DIVMTQSPLSLPVTPGEPASISC (SEQ ID NO:146);

[00372] b) FR2 VL: WYLQKPGQSPQLLIY (SEQ ID NO:147);

[00373] c) FR3 VL: GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC (SEQ ID NO:148); It is

[00374] d) FR4 VL: FGQGTKVEIK (SEQ ID NO:149).

[00375] Non-limiting examples of human consensus VL framework regions suitable for use with VL CDRs as described herein include (subgroup III consensus):

[00376] a) FR1 VL: DIVMTQSPDSLAVSLGERATINC (SEQ ID NO:150);

[00377] b) FR2 VL: WYQQKPGQPPKLLIY (SEQ ID NO:151);

[00378] c) FR3 VL: GVPDRFSGSGSGTDFTLTISSLQAEDFAVYYC (SEQ ID NO:152); It is

[00379] d) FR4 VL: FGQGTKVEIK (SEQ ID NO:153).

[00380] Non-limiting examples of human consensus VL framework regions suitable for use with VL CDRs as described herein include (subgroup IV consensus):

[00381] a) FR1 VL: DIVMTQSPDSLAVSLGERATINC (SEQ ID NO:154);

[00382] b) FR2 VL: WYQQKPGQPPKLLIY (SEQ ID NO:155);

[00383] c) FR3 VL: GVPDRFSGSGSGTDFTLTISSLQAEDFAVYYC (SEQ ID NO:156); It is

[00384] d) FR4 VL: FGQGTKVEIK (SEQ ID NO:157). Formulations

[00385] When carrying out a method of the present invention for treating an alloimmune disorder or an autoimmune disorder, an anti-C1s antibody can be administered to an individual using any convenient means capable of resulting in the desired therapeutic effect. For example, an anti-C1s antibody can be formulated into pharmaceutical compositions by combination with appropriate pharmaceutically acceptable carriers, pharmaceutically acceptable diluents or other pharmaceutically acceptable excipients and can be formulated into preparations in solid, semisolid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants and aerosols.

[00386] In pharmaceutical dosage forms, an anti-C1s antibody can be administered in the form of its pharmaceutically acceptable salts, or it can also be used alone or in appropriate association, as well as in combination, with other pharmaceutically active compounds. The following methods and excipients are merely exemplary and are not limiting in any way.

[00387] For oral preparations, an anti-C1s antibody can be used alone or in combination with appropriate additives to make tablets, powders, granules or capsules, for example, with conventional additives such as lactose, mannitol, corn starch or starch potato; with binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrants such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants such as talc or magnesium stearate; and, if desired, with diluents, buffering agents, wetting agents, preservatives and flavoring agents.

[00388] An anti-C1s antibody can be formulated into preparations for injection by dissolving, suspending or emulsifying the antibody in an aqueous or non-aqueous solvent, such as vegetable oils or the like, propylene glycol, synthetic aliphatic acid glycerides, organic esters injectables (e.g. ethyl oleate), esters of higher aliphatic acids or propylene glycol; and, if desired, with conventional additives, such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactic Ringer's solution, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose) and the like. Furthermore, the pharmaceutical composition of the present invention may comprise additional agents, such as dopamine or psychopharmacological drugs, depending on the intended use of the pharmaceutical composition.

[00389] Pharmaceutical compositions comprising an anti-C1s antibody are prepared by mixing an anti-C1s antibody of the desired degree of purity with optional physiologically acceptable carriers, other excipients, stabilizers, surfactants, buffers and/or tonicity agents. Acceptable carriers, other excipients and/or stabilizers are non-toxic to receptors at the dosages and concentrations employed, and include buffers, such as phosphate, citrate and other organic acids; antioxidants, including ascorbic acid, glutathione, cysteine, methionine and citric acid; preservatives (such as ethanol, benzyl alcohol, phenol, m-cresol, p-chloro-m-cresol, methyl or propyl parabens, benzalkonium chloride or combinations thereof); amino acids, such as arginine, glycine, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenylalanine, tyrosine, tryptophan, methionine, serine, proline and combinations thereof; monosaccharides, disaccharides and other carbohydrates; low molecular weight polypeptides (less than about 10 residues); proteins, such as gelatin or serum albumin; chelating agents such as EDTA; sugars, such as trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine, galactosamine and neuraminic acid; and/or non-ionic surfactants such as Tween, Brij Pluronics, Triton-X or polyethylene glycol (PEG).

[00390] The pharmaceutical composition may be in a liquid form, a lyophilized form, or a liquid form reconstituted from a lyophilized form, wherein the lyophilized preparation must be reconstituted with a sterile solution prior to administration. The standard procedure for reconstituting a lyophilized composition is to add back a volume of pure water (typically equivalent to the volume removed during lyophilization); however, solutions comprising antibacterial agents can be used to produce pharmaceutical compositions for parenteral administration; see also Chen (1992) Drug Dev Ind Pharm 18, 1311-54.

[00391] Examples of antibody concentrations in a pharmaceutical composition can range from about 1 mg/mL to about 200 mg/mL or from about 50 mg/mL to about 200 mg/mL or from about 150 mg/mL mL to about 200 mg/mL.

[00392] An aqueous formulation of an anti-C1s antibody can be prepared in a pH-buffered solution, for example, at pH ranging from about 4.0 to about 7.0 or from about 5.0 to about 6.0, or alternatively about 5.5. Examples of buffers that are suitable for a pH within this range include phosphate, histidine, citrate, succinate, acetate and other organic acid buffers. The buffer concentration may be from about 1 mM to about 100 mM, or from about 5 mM to about 50 mM, depending, for example, on the buffer and the desired tonicity of the formulation.

[00393] A tonicity agent can be included in the antibody formulation to modulate the tonicity of the formulation. Exemplary tonicizing agents include sodium chloride, potassium chloride, glycerin and any component of the group of amino acids, sugars, as well as combinations thereof. In some embodiments, the aqueous formulation is isotonic, although hypertonic or hypotonic solutions may be suitable. The term "isotonic" denotes a solution having the same tonicity as some other solution with which it is compared, such as saline or physiological saline. Tonicizing agents can be used in an amount of about 5 mM to about 350 mM, for example, in an amount of 100 mM to 350 mM.

[00394] A surfactant can also be added to the antibody formulation to reduce aggregation of the formulated antibody and/or minimize the formation of particles in the formulation and/or reduce adsorption. Examples of surfactants include polyoxyethylene sorbitan fatty acid esters (Tween), polyoxyethylene alkyl ethers (Brij), alkylphenylpolyoxyethylene ethers (Triton-X), polyoxyethylene-polyoxypropylene copolymer (Poloxamer, Pluronic), and sodium dodecyl sulfate ( SDS). Examples of suitable polyoxyethylene sorbitan fatty acid esters are polysorbate 20, (sold under the trade name Tween 20™) and polysorbate 80 (sold under the trade name Tween 80™). Examples of suitable polyethylene-polypropylene copolymers are those sold under the names Pluronic® F68 or Poloxamer 188™. Examples of suitable alkyl polyoxyethylene ethers are those sold under the trademark Brij™. Exemplary surfactant concentrations can range from about 0.001% to about 1% w/v.

[00395] A lyoprotectant can also be added in order to protect the labile active ingredient (e.g., a protein) against destabilizing conditions during the lyophilization process. For example, known lyoprotectants include sugars (including glucose and sucrose); polyols (including mannitol, sorbitol and glycerol); and amino acids (including alanine, glycine and glutamic acid). Lyoprotectants can be included in an amount of about 10 mM to 500 nM.

[00396] In some embodiments, a formulation includes an anti-C1s antibody and one or more of the agents identified above (e.g., a surfactant, a buffer, a stabilizer, a tonicity agent) and is essentially free of one or more plus preservatives, such as ethanol, benzyl alcohol, phenol, m-cresol, p-chloro-m-cresol, methyl or propyl parabens, benzalkonium chloride and combinations thereof. In other embodiments, a preservative is included in the formulation, for example, in concentrations ranging from about 0.001 to about 2% (w/v).

[00397] For example, a formulation may be a liquid or lyophilized formulation suitable for parenteral administration and may comprise: about 1 mg/mL to about 200 mg/mL of an anti-C1s antibody; about 0.001% to about 1% of at least one surfactant; about 1 mM to about 100 mM of a buffer; optionally, about 10 mM to about 500 mM of a stabilizer; and about 5 mM to about 350 mM of a tonicity agent; and has a pH of about 4.0 to about 7.0.

EFFECTIVENESS MONITORING METHODS

[00398] The present invention provides a method of monitoring the effectiveness of a method of treating an alloimmune disorder or autoimmune disorder of the present invention. The method generally involves: a) detecting the level of autoantibodies or alloantibodies; and/or b) detect the number of autoreactive or alloreactive B cells; and/or c) detecting the level of a cytokine(s) produced or modulated by a B cell in a biological sample obtained from an individual who has undergone treatment with a method of the present invention for treating a disorder alloimmune or autoimmune disorder. A change (e.g., a decrease) in one or more of: a) the level of autoantibodies or alloantibodies; b) the number of autoreactive or alloreactive B cells; and c) the level of a cytokine(s) produced by or modulated by a B cell, compared to a pre-treatment level or compared to a level or number in a sample collected at an earlier time point, indicates effectiveness of the treatment. In some cases, detection is quantitative.

[00399] In some cases, a method of monitoring the effectiveness of a treatment method of the present invention comprises: a) detecting the level of autoantibodies or alloantibodies in a biological sample initially obtained from an individual; and b) detect the level of autoantibodies or alloantibodies in a biological sample obtained at a second time from the individual. The second moment is after the first moment. When the level of autoantibodies or alloantibodies in the biological sample collected in the second moment is lower than the level of autoantibodies or alloantibodies in the biological sample collected in the first moment, the effectiveness of the treatment is indicated. For alloantibodies (e.g., anti-HLA antibodies), an isotype switch from an alloantibody that binds to C1q to an alloantibody that does not bind to C1q would also demonstrate treatment efficacy. In some cases, a method of monitoring the effectiveness of a treatment method of the present invention comprises: a) detecting the isotype of the autoantibody or alloantibody in a biological sample initially obtained from an individual; and b) detect the isotype of the autoantibody or alloantibody in a biological sample obtained at a second time from the individual.

[00400] In some cases, a method of monitoring the effectiveness of a treatment method of the present invention comprises: a) detecting the level (e.g., determining the number) of autoreactive B cells or alloreactive B cells in a biological sample initially obtained from an individual; and b) detect the level (for example, determine the number) of autoreactive B cells or alloreactive B cells in a biological sample obtained at a second time from the individual. The second moment is after the first moment. When the level of autoreactive B cells or alloreactive B cells in the biological sample collected at the second time point is lower than the level of autoreactive B cells or alloreactive B cells in the biological sample collected at the first time point, treatment efficacy is indicated.

[00401] In some cases, a method of monitoring the effectiveness of a treatment method of the present invention comprises: a) detecting the level of a cytokine(s) produced or modulated by a B cell in a sample biological obtained initially from an individual; and b) detect the level of a cytokine(s) produced or modulated by a B cell in a biological sample obtained at a second time from the individual. The second moment is after the first moment. Where the level of cytokine(s) produced by a B cell or modulated by a B cell in the biological sample collected at the second time point is altered compared to the level of cytokine(s) produced by a B cell or modulated by a B cell in the biological sample collected at the first moment, the effectiveness of the treatment is indicated. For example, where the level of pro-inflammatory cytokine(s) produced by a B cell in the biological sample collected at the second time point is lower than the level of pro-inflammatory cytokine(s) produced (s) by a B cell in the biological sample collected at the first time point, the effectiveness of the treatment is indicated. Cytokines produced by B cells include, for example, pro-inflammatory cytokines such as IL-2, IL-4, IL-6, IL-12, IFN-Y and TNF-α; and immunosuppressive cytokines, such as IL-10 and TGF-β.

[00402] Suitable biological samples include, for example, blood, serum, plasma, etc.

[00403] In some cases, the first time is prior to the start of treatment with a method of the present invention to treat an alloimmune disorder or autoimmune disorder; and the second time is after the start of treatment with a method of the present invention to treat an alloimmune disorder or autoimmune disorder. In some cases, the first time is prior to the initiation of treatment with a method of the present invention to treat an alloimmune disorder or autoimmune disorder; and the second moment is from 2 days to 6 months (for example, from 2 days to 7 days, from 1 week to 2 weeks, from 2 weeks to 4 weeks, from 1 month to 2 months, from 2 months to 3 months, 3 months to 4 months, 4 months to 5 months, or 5 months to 6 months) after starting treatment with a method of the present invention for treating an alloimmune disorder or autoimmune disorder.

[00404] In some cases, the first time is after the start of treatment with a method of the present invention to treat an alloimmune disorder or autoimmune disorder. For example, in some cases, the earliest time point is 2 days to 6 months (e.g., 2 days to 7 days, 1 week to 2 weeks, 2 weeks to 4 weeks, 1 month to 2 months, 2 months to 3 months, 3 months to 4 months, 4 months to 5 months, or 5 months to 6 months) after starting treatment with a method of the present invention to treat an alloimmune disorder or autoimmune disorder; and the second moment is from 2 days to 6 months (for example, from 2 days to 7 days, from 1 week to 2 weeks, from 2 weeks to 4 weeks, from 1 month to 2 months, from 2 months to 3 months, from 3 months to 4 months, from 4 months to 5 months, or from 5 months to 6 months) after the first moment.

[00405] Methods for determining the level of autoantibodies or alloantibodies are known in the art and any known method can be used. Examples of suitable methods include immunological methods such as ELISA, LFIA, DIA, FIA, CLIA, CIA, MIA, RIA and the like. For example, a detectably labeled autoantigen or alloantigen can be used in an assay to detect an autoantibody or alloantibody, respectively. The autoantibody present in a biological sample obtained from an individual being treated can be immobilized and the detectably labeled autoantigen can be brought into contact with the immobilized autoantibody forming a complex, where the presence or amount of detectable marker indicates the presence or amount of autoantibody in the biological sample. Similarly, the alloantibody present in a biological sample obtained from an individual being treated can be immobilized and the detectably labeled alloantigen can be brought into contact with the immobilized alloantibody forming a complex, where the presence or amount of detectable marker indicates the presence or amount of alloantibody in the biological sample.

[00406] Methods of determining the level (e.g., the number) of autoreactive B cells or alloreactive B cells are known in the art, and any known method can be used. Examples of suitable methods include flow cytometry, immunofluorescence, enzyme-linked immunospot assay (ELISPOT), etc. The level (e.g., number) of autoreactive B cells or alloreactive B cells is determined in a sample obtained from an individual, where the sample may include, for example, a biopsy sample of tissue, blood, or bone marrow.

[00407] Methods of determining the level of a cytokine produced or modulated by a B cell are known in the art, and any known method can be used. Examples of suitable methods include, for example, an ELISA assay. Suitable Subjects for Treatment

[00408] A variety of hosts (where the term "host" is used interchangeably herein with the terms "subject," "individual" and "patient") is treatable according to the targeted methods. Generally such hosts are "mammals," where that term is used broadly to describe organisms that fall within the mammalian class, including the orders carnivorous (e.g., cats), herbivores (e.g., cattle, horses, and sheep). daughters), omnivores (e.g., dogs, goats, and pigs), rodents (e.g., rats, guinea pigs, and rats), and primates (e.g., humans, chimpanzees, and monkeys). In some embodiments, the host is an individual that has a complement system, such as a mammal, fish, or invertebrate. In some embodiments, the host is a mammalian, fish or invertebrate companion animal, agricultural animal, working animal, zoo animal or laboratory animal that contains a complement system. In some embodiments, the host is a human being.

[00409] Individuals suitable for treatment using a method of the present invention to treat an autoimmune disorder include individuals with an autoimmune disorder mediated by autoantibodies. Suitable individuals for treatment using a method of the present invention to treat an autoimmune disorder include individuals who have a disorder (e.g., diagnosed as having a disorder), such as Addison's disease, age-related macular degeneration, alopecia, autoimmune hepatitis (e.g., example, autoimmune hepatitis associated with hepatitis B virus infection; autoimmune hepatitis associated with hepatitis C virus infection), autoimmune hemolytic anemia, autoimmune skin diseases, autoimmune thyroid disease, bullous pemphigus, celiac disease, cold agglutinin disease, dermatomyositis, type 1 diabetes mellitus, Grave's disease, Goodpasture's syndrome, Hashimoto's disease, hypoparathyroidism, hypopituitarism, hypothyroidism, idiopathic thrombocytopenic purpura, inflammatory bowel disease (e.g., Crohn's disease, ulcerative colitis), multiple sclerosis, myasthenia gravis, myocarditis, neuromyelitis optica, pemphigus vulgaris, pemphigus foliaceus, polymyositis, psoriasis, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, uveitis and granulomatosis of Wegener and poly/dermatomyositis.

[00410] In some cases, the individual has been previously treated with a treatment regimen for an autoimmune disorder and has not responded to the treatment. In some cases, the individual was previously treated with a treatment regimen for an autoimmune disorder and relapsed, for example, the autoimmune disorder reappeared.

[00411] In some cases, an individual who is suitable for treatment with a method of the present invention has a disease selected from age-related macular degeneration, Alzheimer's disease, amyotrophic lateral sclerosis, anaphylaxis, argyrophilic grain dementia, arthritis (e.g. , rheumatoid arthritis), asthma, atherosclerosis, atypical hemolytic-uremic syndrome, autoimmune diseases, autoimmune hemolytic anemia, Barraquer-Simons syndrome, Behçet's disease, British-type amyloid angiopathy, bullous pemphigus, Buerger's disease, C1q nephropathy, cancer, catastrophic antiphospholipid syndrome, cerebral amyloid angiopathy, cold agglutinin disease, corticobasal degeneration, Creutzfeldt-Jakob disease, Crohn's disease, cryoglobulinemic vasculitis, dementia pugilistica, dementia with Lewy bodies (DLB), diffuse neurofibrillary tangles with calcification, discoid lupus erythematosus, Down syndrome, focal segmental glomerulosclerosis, formal thought disorder, frontotemporal dementia (FTD), frontotemporal dementia with parkinsonism linked to chromosome 17, frontotemporal lobar degeneration, Gerstmann-Straussler-Scheinker disease, Guillain syndrome- Barré, Hallervorden-Spatz disease, hemolytic uremic syndrome, hereditary angioedema, hypophosphatasis, idiopathic pneumonia syndrome, immune complex diseases, inclusion body myositis, infectious diseases (e.g., disease caused by bacteria (e.g., Neisseria meningitidis or Streptococcus) viruses (e.g., human immunodeficiency virus (HIV)), or other infectious agents), inflammatory disease, ischemia/reperfusion injury, mild cognitive impairment, immunothrombocytopenic purpura (ITP), molybdenum (MoCD) type A, membranoproliferative glomerulonephritis (MPGN) I, membranoproliferative glomerulonephritis (MPGN) II (dense deposit disease), membranous nephritis, multi-infarct dementia, lupus (e.g., systemic lupus erythematosus (SLE)), glomerulonephritis te, Kawasaki disease, multifocal motor neuropathy, multiple sclerosis, multiple system atrophy, myasthenia gravis, myocardial infarction, myotonic dystrophy, neuromyelitis optica, Niemann-Pick disease type C, non-Guamanian motor neuron disease with neurofibrillary tangles , Parkinson's disease, Parkinson's disease with dementia, paroxysmal nocturnal hemoglobinuria, pemphigus vulgaris, Pick's disease, post-encephalitis parkinsonism, polymyositis, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, progressive supranuclear palsy, psoriasis, sepsis, Shiga toxin coli (STEC)-HuS, spinal muscular atrophy, subacute sclerosing panencephalitis, stroke, tangle-only dementia, transplant rejection, vasculitis (e.g., ANCA-associated vasculitis), Wegner's granulomatosis, sickle cell disease, cryo - globulinemia, mixed cryoglobulinemia, essential mixed cryoglobulinemia, mixed cryoglobulinemia type II, mixed cryoglobulinemia type III, nephritis, drug-induced thrombocytopenia, lupus nephritis, bullous pemphigus, epidermolysis bullosa acquisita, delayed hemolytic transfusion reaction, urticarial vasculitis syndrome hypocomplementemic, pseudophakic bullous keratopathy and platelet refractoriness.

[00412] Individuals suitable for treatment using a method of the present invention to treat an alloimmune disorder include individuals who have received a donor organ or tissue, where such individuals are referred to as organ or tissue recipients. Subjects suitable for treatment using a method of the present invention to treat an alloimmune disorder include subjects who are about to receive (e.g., who are scheduled to receive, who are on a waiting list to receive, etc.) a donor organ or tissue. , where such individuals are referred to as potential organ or tissue recipients. Allografted organs and tissues include, but are not limited to, a kidney, a liver, a pancreas, a heart, a lung, skin, blood tissue (including whole blood, red blood cells, white blood cells, umbilical cord blood, and the like) , where the blood tissue may comprise an isolated population of blood cells (buffy coat, red blood cells, platelets, lymphocytes, T cells, B cells or some other population), or where the blood tissue comprises a mixed population of cells, intestine thin tissue, an endothelial tissue, a vascular tissue (e.g., a blood vessel), an eye, a stomach, a thymus, bone, bone marrow, cornea, a heart valve, an islet of Langerhans, or a tendon.

EXAMPLES

[00413] The following examples are presented in order to provide those skilled in the art with a complete description and description of how to make and use the present invention and are not intended to limit the scope of what the inventors consider their invention nor are they intended to represent that the Experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. quantities, temperature, etc.), but some errors and experimental deviations must be considered. Unless otherwise indicated, parts are parts by weight, molecular weight is the weight average molecular mass, temperature is in degrees Celsius, and pressure is atmospheric or close to it. Standard abbreviations may be used, e.g. bp, base pair(s); kb, kilobase(s); pL, picoliter(s); s or sec, second(s); min, minute(s); h or hr, hour(s); aa, amino acid(s); kb, kilobase(s); bp, base pair(s); nt, nucleotide(s); i.m., intramuscular(mind); i.p., intraperitoneal(mind); s.c., subcutaneous (mind); and the like.

Example 1 Materials and methods

[00414] Preparation of ELISA plates. High-binding ELISA plates were coated overnight at +4°C with 10 μg/mL endotoxin-free plasma-derived total human IgM (Hu IgM) or with mouse IgG raised against human IgM (MaH). The next day, the plates were washed with phosphate-buffered saline (PBS) and blocked with 2% gelatin solution.

[00415] Complement deposition. Normal human serum was diluted in 2.5% GVB++ buffer for Hu IgM plates or 5% for MaH plates and treated for 15-30 minutes at room temperature with: 1) 20-100 μg/mL of monoclonal antibody from mouse anti-C1s TNT003, a humanized variant of TNT003, anti-C5 antibody, or corresponding isotype controls; 2) 350 μg/mL mouse monoclonal anti-C1s antibody TNT005, a humanized variant of TNT005, or corresponding isotype control; or 3) human serum immunodeleted for C3. The resulting serum solutions were incubated in the corresponding plate for 90 minutes at 37°C. Complement deposition was stopped with 3 washes with PBS at room temperature.

[00416] Coating of human IgM plates with B cell receptor (BCR) agonist. Plates with Hu IgM exposed to normal human serum (NHS) were carefully washed and exposed to 15 μg/mL of F(ab')2 fragment of goat anti-human IgM antibody specific for Fc5μ for 30 minutes at room temperature. Plates were washed three times with PBS.

[00417] Activation of primary B cells. Normal human primary B cells were prestained with Fluo-4 Ca2+ flux reporter according to the manufacturer's protocol. Stained cells were exposed to MaH or Hu IgM-coated plates with complement deposited for 1 hour. Fluo-4 fluorescence was measured on the Spectromax i3 instrument and Ca2+ flux values were determined as the area under the curve.

[00418] Proliferation of primary B cells. Normal primary human B cells pre-stained with carboxyfluorescein succinimidyl ester (CFSE) were incubated on MaH-coated plates with complement deposited for 1 hour and then stimulated with CpG oligode-soxynucleotide (ODN) with TLR9 ligand and maintained in an incubator. with CO2. Eight days after stimulation, cells were fixed with 4% paraformaldehyde and stained with an antibody specific for CD19 conjugated to allophycocyanin (APC). Cells were analyzed by flow cytometry. The proliferative population of cells was identified as the percentage of cells with low CFSE in positive gate selection for intact single CD19.

[00419] C3d, C5b ELISA. Plates coated with Hu IgM or MaH with deposited complement were blocked in 1% casein and stained with rabbit anti-human C3d or rabbit anti-human C5b primary antibody for 1 hour. Then the plates were carefully washed in PBS + TWEEN® 20 nonionic detergent (PBST) and stained with horseradish peroxidase (HRP)-conjugated anti-rabbit antibody for 1 hour. Plates were washed in PBST. The HRP signal was revealed using the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The reaction was stopped after 10 minutes with a low pH solution. C3d or C5b deposition was measured as optical density (OD) absorption at 405 nm in a plate reader and normalized to the levels of an appropriate isotype control.

Results

[00420] The results are represented in FIG. 2A-2D, FIG. 3A-3C, FIG. 4A-4C, and FIG. 5A-5C.

[00421] As shown in FIG. 2A-2D, TNT003, a mouse monoclonal antibody that inhibits human C1s, prevents C3 complement-mediated activation of normal primary human B cells.

[00422] FIG. 2A-2D. (A). ELISA C3d. Deposition of the C3d complement fragment using normal human serum treated with an isotype control (mouse IgG2a), TNT003, or using human serum immunodeleted for C3 (C3dpl) on ELISA plates coated with human IgM. (B). Activation of primary human B cells. Calcium (Ca2+) flux in normal primary human B cells exposed to plaques with deposited complement of (A) activated by the addition of anti-Ig B cell receptor (BCR) agonist. (W). ELISA C3d. Deposition of the C3d complement fragment using normal human serum treated with an isotype control (mouse IgG2a), TNT003, or using human serum immunodeleted for C3 on ELISA plates coated with mouse IgG. (D). Activation of primary human B cells. Ca2+ flow in normal primary human B cells exposed to plaques with deposited complement from (C). Mouse anti-human IgG is used as an immobilized BCR agonist. Values are normalized to isotype-matched controls and are the average of four independent experiments on primary B cells derived from the blood of four separate human donors. Statistics were calculated using one-way ANOVA (Tukey's multiple comparison test).

[00423] As shown in FIG. 3A-3C, a humanized variant of TNT003 ("huTNT003"), which is a humanized IgG4 monoclonal antibody that inhibits human C1s, prevents C3 complement-mediated activation of normal primary human B cells.

[00424] FIG. 3A-3C. (A). ELISA C3d. Deposition of the C3d complement fragment using human serum treated with an isotype control (human IgG4) or a humanized variant of TNT003 on ELISA plates coated with B cell receptor (BCR) agonist mouse IgG. (B). Activation of primary human B cells exposed to deposited complement. Ca2+ flux in normal primary human B cells exposed to plaques (A). (W). Proliferation of primary human B cells exposed to deposited complement. Normalized ratio of CD19+ CFSE-low B cells exposed to (A) plaques in the presence of the Toll-like receptor 9 (TLR9) agonist CpG oligodeoxynucleotide (ODN) on day 8 post-stimulation. Values are normalized to isotype-matched controls and are the average of five independent experiments on primary B cells derived from the blood of five separate human donors. Statistics were calculated using one-way ANOVA (Tukey's multiple comparison test).

[00425] As shown in FIG. 4A-4C, the C1s inhibitor (a humanized variant of TNT003), but not the C5 inhibitor antibody, prevents C3 complement-mediated activation of normal human primary B cells.

[00426] FIG. 4A-4C. (A). ELISA C3d. Deposition of the C3d complement fragment using human serum treated with an isotype control, a humanized variant of TNT003, C5 inhibitory antibody (αC5), or C3-deleted serum on ELISA plates coated with B cell receptor agonist mouse IgG (BCR). (B). ELISA C5b. Deposition of complement C5d using human serum treated with an isotype control, humanized variant of TNT003, C5 inhibitory antibody (αC5), or serum deleted for C3 on ELISA plates coated with B cell receptor (BCR) agonist mouse IgG . (W). Activation of primary human B cells exposed to deposited complement. Ca2+ flux in normal primary human B cells exposed to plaques (A, B). Values are normalized to isotype-matched controls and are the average of seven independent experiments on primary B cells derived from the blood of seven separate human donors. Statistics were calculated using one-way ANOVA (Tukey's multiple comparison test). The statistics presented are the comparison with an appropriate isotype control. The difference in B cell response between the humanized point-deleted variant for TNT003 and C3 is statistically non-significant (ns).

[00427] As shown in FIG. 5A-5C, C1s inhibitory antibodies with distinct modes of action, but not the C5 inhibitory antibody, prevent C3 complement-mediated activation of normal human primary B cells.

[00428] FIG. 5A-5C. (A). ELISA C3d. Deposition of the C3d complement fragment using human serum treated with mouse isotype controls, TNT005 (a mouse IgG2a monoclonal C1s inhibitory antibody with distinct mode of action), human isotype controls, humanized variant of TNT003, a of TNT005 humanized IgG4, C5 inhibitory antibody (αC5) or C3-deleted serum on ELISA plates coated with B cell receptor (BCR) agonist mouse IgG. (B). ELISA C5b. Deposition of complement C5b using human serum treated as in (A). (W). Activation of primary human B cells exposed to deposited complement. Ca2+ flux in normal primary human B cells exposed to plaques (A, B). Values are normalized to isotype-matched controls and are the average of three independent experiments on primary B cells derived from the blood of three separate human donors. Statistics were calculated using one-way ANOVA (Tukey's multiple comparison test). The statistics presented are the comparison with an appropriate isotype control. The difference in B cell response between the points of the humanized TNT003 and C3-deleted variant, between the humanized TNT005 ("huTNT005") and C3-deleted variant, and between the humanized TNT005 and C3-deleted variant is statistically not significant(ns).

[00429] Although the present invention has been described with reference to specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. Furthermore, many modifications can be made to adapt a particular situation, material, composition of matter, process or step or steps of the process to the objective, essence and scope of the present invention. All such modifications are intended to be within the scope of the claims appended hereto.

Claims (27)

1. Use of an antibody that specifically binds to complement component 1s (C1s), characterized in that it is for the preparation of a medicine or pharmaceutical composition for reducing the level of autoantibody or alloantibody titers in an individual affected by an autoimmune or alloimmune disorder, wherein the antibodies comprise: (i) light chain complementarity determining regions (CDRs) of a light chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 8 and heavy chain CDRs of a heavy chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 7; or (ii) light chain CDRs of a light chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 94 and heavy chain CDRs of a heavy chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 93. 2. Use, according to claim 1, characterized by the fact that it comprises monitoring the effectiveness of the administration of said antibody, comprising detecting a level of autoantibodies or alloantibodies in a biological sample obtained from the individual. 3. Use according to claim 2, characterized by the fact that it comprises adjusting the dose of the anti-C1s antibody based on the detected level. 4. Use according to any one of claims 1 to 3, characterized by the fact that the individual is a human being. 5. Use according to any one of claims 1 to 4, characterized by the fact that the anti-C1s antibody is humanized. 6. Use according to claim 5, characterized by the fact that the anti-C1s antibody comprises a humanized VL framework region. 7. Use according to claim 5, characterized in that the anti-C1s antibody comprises a humanized VH framework region. 8. Use according to claim 5, characterized in that the anti-C1s antibody comprises a humanized VL framework region and a humanized VH framework region. 9. Use of an antibody that specifically binds to complement component 1s (C1s), characterized by the fact that it is for the preparation of a medicine or pharmaceutical composition for reducing the proliferation of B cells in an individual affected by an autoimmune disorder or alloimmune, wherein the antibodies comprise: (i) light chain complementarity determining regions (CDRs) of a light chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 8 and heavy chain CDRs of a variable region of an antibody heavy chain comprising the amino acid sequence SEQ ID NO: 7; or (ii) light chain CDRs of a light chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 94 and heavy chain CDRs of a heavy chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 93. 10. Use, according to claim 9, characterized by the fact that it comprises monitoring the effectiveness of the administration of said antibody, comprising detecting a level of B cell proliferation in a biological sample obtained from the individual. 11. Use according to claim 10, characterized by the fact that it comprises adjusting the dose of the anti-C1s antibody based on the level detected. 12. Use according to any one of claims 9 to 11, characterized by the fact that the individual is a human being. 13. Use according to any one of claims 9 to 12, characterized by the fact that the anti-C1s antibody is humanized. 14. Use according to claim 13, characterized in that the anti-C1s antibody comprises a humanized VL framework region. 15. Use according to claim 13, characterized in that the anti-C1s antibody comprises a humanized VH framework region. 16. Use according to claim 13, characterized in that the anti-C1s antibody comprises a humanized VL framework region and a humanized VH framework region. 17. Use of an antibody that specifically binds to component 1s (C1s) of complement, characterized by the fact that it is for the preparation of a medicine or pharmaceutical composition for reducing the activation of B cells in an individual affected by an autoimmune disorder or alloimmune, wherein the antibodies comprise: (i) light chain complementarity determining regions (CDRs) of a light chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 8 and heavy chain CDRs of a variable region of an antibody heavy chain comprising the amino acid sequence SEQ ID NO: 7; or (ii) light chain CDRs of a light chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 94 and heavy chain CDRs of a heavy chain variable region of an antibody comprising the amino acid sequence SEQ ID NO: 93. 18. Use according to claim 17, characterized by the fact that the individual is a human being. 19. Use according to claim 17 or 18, characterized by the fact that the anti-C1s antibody is humanized. 20. Use according to claim 19, characterized in that the anti-C1s antibody comprises a humanized VL framework region. 21. Use according to claim 20, characterized in that the anti-C1s antibody comprises a humanized VH framework region. 22. Use according to claim 19, characterized in that the anti-C1s antibody comprises a humanized VL framework region and a humanized VH framework region. 23. Use according to any one of claims 1 to 22, characterized in that the antibody is administered in an amount of 0.1 mg/kg to 100 mg/kg. 24. Use according to any one of claims 1 to 23, characterized in that the antibody is administered once a day, twice a week, once a week, every two weeks, or once a month. 25. Use according to any one of claims 1 to 24, characterized in that the antibody is administered intravenously, subcutaneously or intramuscularly. 26. Use according to any one of claims 1 to 25, characterized in that the antibody comprises: a) CDR-H1 comprising the amino acid sequence GFNIKDDYIHWV (SEQ ID NO: 9); CDR-H2 comprising the amino acid sequence IDPADGHTKY (SEQ ID NO: 10); and CDR-H3 comprising the amino acid sequence ARYGYGREVFDY (SEQ ID NO: 11); and b) CDR-L1 comprising the amino acid sequence QSVDYDGDSYMN (SEQ ID NO: 12); CDR-L2 comprising the amino acid sequence DASNLESGIP (SEQ ID NO: 13); and CDR-L3 comprising the amino acid sequence QQSNEDPWT (SEQ ID NO: 14). 27. Use according to any one of claims 1 to 25, characterized in that the antibody comprises: a) CDR-H1 comprising the amino acid sequence NYAMS (SEQ ID NO: 95); CDR-H2 comprising the amino acid sequence TISSGGSHTYYLDSVKG (SEQ ID NO: 96); and CDR-H3 comprising the amino acid sequence LFTGYAMDY (SEQ ID NO: 97); and b) CDR-L1 comprising the amino acid sequence TASSSVSSSYLH (SEQ ID NO: 98); CDR-L2 comprising the amino acid sequence STSNLAS (SEQ ID NO: 99); and CDR-L3 comprising the amino acid sequence HQYYRLPPIT (SEQ ID NO: 92).