IL324305A - Antibodies directed against xcl1 and methods of using them - Google Patents

Description

30574 WO ANTIBODIES TARGETING XCL1 AND METHODS OF USING THE SAME SEQUENCE LISTING FILE [ 0001 ] The present application is being filed along with a Sequence Listing in ST.26 XML format . The Sequence Listing is provided as a file titled " 30574_SequenceListing " created May 6 , 2024 , and is 43 kilobytes in size . The Sequence Listing information in the ST.26 XML format is incorporated herein by reference in its entirety .

FIELD OF THE INVENTION [ 0002 ] The present invention is in the field of medicine . More particularly , the present invention relates to antibodies directed against lymphotactins , in particular the human chemokines XCL1 and XCL2 ; pharmaceutical compositions comprising such antibodies ; and methods of using such antibodies . The antibodies disclosed herein bind and neutralize XCL1 and XCL2 . The antibodies and methods of the present invention are expected to be useful in the field of autoimmune and chronic inflammatory diseases ( collectively referred to herein as , immune - mediated diseases ) , particularly diseases such as alopecia areata , vitiligo , ulcerative colitis ( UC ) , rheumatoid arthritis ( RA ) , multiple sclerosis ( MS ) , type 1 diabetes , psoriasis , asthma , and the like , including treatment thereof .

BACKGROUND OF THE INVENTION [ 0003 ] XCL1 and XCL2 , also known as lymphotactin - a and u0000- , respectively , are the sole members of the C chemokine / y chemokine family . XCL1 is expressed and secreted by various immune cells , including activated CD8 + T cells , CD4 + T cells , natural killer ( NK ) cells , thymic medullary epithelial cells , and others . XCL2 is a paralog of XCL1 and is thought to have arisen from a gene duplication , having a nearly - identical amino acid sequence to XCL1 ( differing by only two amino acids in its N- terminal region from XCL1 ) . XCL1 and XCL2 are thought to have similar functions as a result of this structural similarity . [ 0004 ] Unlike most other chemokines , which have four cysteine residues near their amino termini capable of forming two disulfide bonds , XCL1 and XCL2 have just two such cysteines , which form a single disulfide bond . This allows the chemokine to be metamorphic , interconverting between a first configuration having a canonical monomeric , α - helix - containing chemokine fold , and a second , primarily u0000 - sheet form . This conversion between forms typically occurs several times per second . The 30574 WO monomeric α - helical form is capable of binding and activating the receptor XCR1 to induce cell migration . The a - helical form undergoes a conformational change to the u0000 - sheet form , which can exist in a monomeric form but favors dimerization . When in the dimeric or u0000 - sheet configuration , XCL1 is inactive for XCR1 binding , and the dimer can adhere to glycosaminoglycan ( GAG ) on cell surfaces . The monomeric α - helical form does not bind GAG . See Lei and Takahama , Microbes and Infection ( 2012 ) 262-267 ; Fox et al . , ACS Chem . Biol . 10 ( 11 ) ( 2015 ) 2580-2588 . The frequent conversion between these forms , particularly given the inactivity of the dimeric and u0000 - sheet form and its tendency to aggregate at the cell surface on GAG , complicates the design of anti - XCL1 and anti - XCLantibodies , particularly as therapeutic agents , as an antibody that can bind the dimeric form of the chemokine could primarily bind the inactive form of the chemokine , leading to unknown properties with regard to pharmacokinetics , immunogenicity , and efficacy . [ 0005 ] Autoimmune diseases are a form of immune - mediated diseases that arise from the body's production of an immune response against its own tissue . These conditions are often chronic , and can be debilitating and even life - threatening . Current FDA approved treatments for immune - mediated diseases include corticosteroids , often used to treat acute inflammation , and bioproducts targeting αFNT , interleukin - 12 , -17 and -23 . Although these treatments have demonstrated efficacy in reducing symptoms for a subset of patients , a percentage of patients remain nonresponsive or experience a loss of response to the currently available treatments . Thus , there remains a need for additional therapies targeting different human targets and pathways for the treatment of immune - mediated diseases . [ 0006 ] A subset of autoimmune diseases has been associated with overactivity of and tissue damage by CD8 + T cells when these cells have migrated to certain sites in the body , including alopecia areata , vitiligo , UC , type 1 diabetes , psoriasis , RA , MS , and asthma , though many of the specifics around the mechanisms of CD8 + T cell activation underlying these conditions have not been elucidated . For example , roles specific to CD8 + T cell activation by dendritic cells ( DCs ) have generally been elucidated in the field of oncology , rather than immunology , such as in studies of checkpoint antagonists . This is true of the subset of DCs known as conventional dendritic cell 1 ( CDC1 ) ; see foe example Bowman - Kirigin et al . , " The Conventional Dendritic Cell 1 Subset Primes CD8 + T Cells and Traffics Tumor Antigen to Drive Antitumor Immunity in the Brain , " Cancer Immunol Res , 11 ( 1 ) ( 2023 ) p . 20-37 . 30574 WO [ 0007 ] While XCL1 antibodies exist as research tools for detection and visualization of XCL1 , to date there are no approved or known clinical candidates for XCL1 - specific , antagonistic antibody therapeutics . Thus , there remains an unmet need for antibodies , pharmaceutical compositions , and methods which target human XCL1 useful for the treatment of immune - mediated diseases such as RA , MS , asthma , psoriasis , UC , type 1 diabetes , alopecia areata , vitiligo and the like . There is a need for XCL1 antibodies which possess good therapeutic characteristics , including being potent neutralizers ( i.e. , antagonists ) of human XCL1 . Advantageous properties of such antibodies may include high binding affinity for human XCL1 and low nonspecific binding , particularly to other chemokines , which may have similar sequence and / or structure to XCL1 . Such XCL1 antibodies should also possess therapeutically acceptable pharmacokinetics ( Pk ) profile and demonstrate low immunogenicity . Such XCL1 antibodies should also be amendable to commercial manufacturing , including high levels of solubility and low levels of aggregation . The present disclosure provides XCL1 antibodies that address one or more of these needs for use in the treatment of immune - mediated diseases . [ 0008 ] Accordingly , in certain embodiments , the present invention provides antibodies directed against human XCL1 . The antibodies disclosed herein also bind to and neutralize human XCL2 .

SUMMARY OF THE INVENTION [ 0009 ] According to some embodiments , the antibodies of the present invention are antagonistic to human XCL1 , and in some embodiments , antagonistic to human XCL2 . In one aspect , described herein is an antibody that binds human XCL1 which includes a heavy chain variable region ( HCVR ) and a light chain variable region ( LCVR ) . The HCVR includes complementarity determining regions ( CDRs ) HCDR1 , HCDR2 , and HCDR3 . The LCVR comprises CDRs LCDR1 , LCDR2 , and LCDR3 . The HCDR1 is or includes SEQ ID NO : 10. The HCDR2 is or includes SEQ ID NO : 11. The HCDRis or includes SEQ ID NO : 12 , wherein Z is selected from Q and E. The LCDR1 is or includes SEQ ID NO : 5 , wherein X is selected from S and D. The LCDR2 is or includes SEQ ID NO : 8. The LCDR3 , is or includes SEQ ID NO : 9 . [ 0010 ] In one embodiment , Z is E. In another embodiment , Z is Q. 30574 WO [ 0011 ] In one embodiment , X is D. In another embodiment , X is S. [ 0012 ] In one embodiment HCDR1 is or includes SEQ ID NO : 10 ; HCDR2 is or includes SEQ ID NO : 11 ; HCDR3 is or includes SEQ ID NO : 13 ; LCDR1 is or includes SEQ ID NO : 6 ; LCDR2 is or includes SEQ ID NO : 8 ; and LCDR3 is or includes SEQ ID NO : 9 . [ 0013 ] In one embodiment , the LCVR is or includes SEQ ID NO : 17 and the HCVR is or includes SEQ ID NO : 15 , wherein Xa is selected from Q and pyroglutamic acid . In one embodiment , Xa is Q. [ 0014 ] In one embodiment , the antibody has a heavy chain ( HC ) and a light chain ( LC ) , wherein the amino acid sequence of the HC is or includes SEQ ID NO : 19 , wherein X1 is selected from Q and pyroglutamic acid ; X2 is G or absent ; and X3 is K or absent ; and the amino acid sequence of the LC is or includes SEQ ID NO : 21. In one embodiment , X1 is Q ; X2 is G , and X3 is K. [ 0015 ] In one embodiment , HCDR1 is or includes SEQ ID NO : 10 ; HCDR2 is or includes SEQ ID NO : 11 ; HCDR3 is or includes SEQ ID NO : 14 ; LCDR1 is or includes SEQ ID NO : 7 ; LCDR2 is or includes SEQ ID NO : 8 ; and LCDR3 is or includes SEQ ID NO : 9 . [ 0016 ] In one embodiment , the amino acid sequence of the LCVR is or includes SEQ ID NO : 18 and the amino acid sequence of the HCVR is or includes SEQ ID NO : 16 , wherein Xa is selected from Q and pyroglutamic acid . In one embodiment , Xa is Q. [ 0017 ] In one embodiment , the antibody has a heavy chain ( HC ) and a light chain ( LC ) . The HC is or includes SEQ ID NO : 20 , wherein X1 is selected from Q and pyroglutamic acid ; X2 is G or absent ; and X3 is K or absent ; and the LC is or includes SEQ ID NO : 22. In one embodiment , X1 is Q ; X2 is G ; and X3 is K. [ 0018 ] In any of the above embodiments , the antibody may be an XCL1 antagonist . [ 0019 ] In any of the above embodiments , the antibody may bind human XCL2 . [ 0020 ] In any of the above embodiments , the antibody may be an XCL2 antagonist . [ 0021 ] In any of the above embodiments , the antibody may bind dimeric XCL1 . The dimeric XCLmay be homodimeric XCL1 . The antibody may bind XCL1 in a u0000 - sheet configuration . The homodimeric XCL1 may be bound to glycosaminoglycan on a surface of a cell . [ 0022 ] In any of the above embodiments , the antibody may bind dimeric XCL2 . The dimeric XCLmay be homodimeric XCL2 . The antibody may bind XCL2 in a u0000 - sheet configuration . The homodimeric XCL2 may be bound to glycosaminoglycan on a surface of a cell . 30574 WO [ 0023 ] In any of the above embodiments , the antibody may bind a heterodimeric complex including one monomer of XCL1 and one monomer of XCL2 . In such an embodiment , the antibody binds at least one amino acid of the XCL1 monomer or at least one amino acid of the XCL2 monomer of the heterodimeric complex . The heterodimeric complex may be bound to glycosaminoglycan on a surface of a cell . [ 0024 ] In any of the above embodiments , the antibody does not bind a chemokine other than XCLand XCL2 . [ 0025 ] In one embodiment , the antibody includes a heavy chain variable region ( HCVR ) having at least 95 % sequence identity to SEQ ID NO : 15 and a light chain variable region ( LCVR ) having at least 95 % sequence identity to SEQ ID NO : 17 . [ 0026 ] In one embodiment , the antibody includes a heavy chain ( HC ) having at least 95 % sequence identity to SEQ ID NO : 19 , and a light chain ( LC ) having at least 95 % sequence identity to SEQ ID NO : 21 . [ 0027 ] In one embodiment , the antibody includes a heavy chain variable region ( HCVR ) having at least 95 % sequence identity to SEQ ID NO : 16 and a light chain variable region ( LCVR ) having at least 95 % sequence identity to SEQ ID NO : 18 . [ 0028 ] In one embodiment , the antibody includes a heavy chain ( HC ) having at least 95 % sequence identity to SEQ ID NO : 20 , and a light chain ( LC ) having at least 95 % sequence identity to SEQ ID NO : 22 . [ 0029 ] In one embodiment , the antibody binds an epitope of XCL1 including one or more amino acids of SEQ ID NO : 27. In one embodiment , the antibody binds two or more amino acids of SEQ ID NO : 27. In an embodiment , the antibody binds three or more amino acids of SEQ ID NO : 27 . [ 0030 ] In one embodiment , the antibody binds one or more amino acids of SEQ ID NO : 29 . [ 0031 ] In one embodiment , the antibody binds an epitope of XCL1 including one or more amino acids of SEQ ID NO : 28. In one embodiment , the antibody binds two or more amino acids of SEQ ID NO : . [ 0032 ] In embodiment , the antibody which binds an epitope of XCL1 including one or more amino acids of SEQ ID NO : 27 and one or more amino acids of SEQ ID NO : 28. In one embodiment , the 30574 WO antibody binds two or more amino acids of SEQ ID NO : 27 and to or more amino acids of SEQ ID NO : 28. In one embodiment , the antibody binds one or more amino acids of SEQ ID NO : 29 . [ 0033 ] In one embodiment , the antibody has a human IgG1 isotype . [ 0034 ] The present disclosure also provides a pharmaceutical composition including an antibody as described in any of the above embodiments and one or more pharmaceutically acceptable carriers , diluents or excipients . [ 0035 ] The present disclosure provides a method of treating alopecia areata , vitiligo , asthma , psoriasis , multiple sclerosis , psoriasis , rheumatoid arthritis , or ulcerative colitis in a subject in need thereof , comprising administering to the subject an effective amount of an antibody or a pharmaceutical composition as described herein . [ 0036 ] The present disclosure provides an antibody or pharmaceutical composition for use in therapy , such as for use in the treatment of alopecia areata , vitiligo , asthma , psoriasis , multiple sclerosis , rheumatoid arthritis , or ulcerative colitis . [ 0037 ] The present disclosure provides for the use of an antibody or a pharmaceutical composition in the manufacture of a medicament for the treatment of alopecia areata , vitiligo , asthma , psoriasis , multiple sclerosis , rheumatoid arthritis , or ulcerative colitis . [ 0038 ] In one aspect , a nucleic acid including a sequence encoding SEQ ID NO : 23 , SEQ ID NO : 24 , SEQ ID NO : 25 , or SEQ ID NO : 26 , is provided . In one aspect , a vector including a first nucleic acid sequence encoding SEQ ID NO : 23 or SEQ ID NO : 25 and a second nucleic acid sequence encoding SEQ ID NO : 24 or SEQ ID NO : 26 is described , as is a cell including such a vector . In one embodiment , a method of producing an antibody including culturing said cell under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium is disclosed . In one embodiment , an antibody produced by culturing said cell under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium is disclosed . [ 0039 ] In one aspect , a protein complex comprising dimeric XCL1 and an antibody bound to dimeric XCL1 is provided . The dimeric XCL1 may be homodimeric XCL1 . The antibody may bind at least one amino acid residue on each monomeric unit of dimeric XCL1 . 30574 WO [ 0040 ] In one aspect , a protein complex comprising dimeric XCL2 and an antibody bound to dimeric XCL2 is provided . The dimeric XCL2 may be homodimeric XCL2 . The antibody may bind at least one amino acid residue on each monomeric unit of dimeric XCL2 . [ 0041 ] In one aspect , a protein complex including a heterodimer made up of a monomeric unit of XCL1 and a monomeric unit of XCL2 , and an antibody bound to the heterodimer , is provided . In this embodiment , the antibody may bind at least one amino acid residue of XCL1 , or at least one amino acid residue of XCL2 , or at least one residue of XCL1 and at least one residue of XCL2 .

DETAILED DESCRIPTION OF THE INVENTION [ 0042 ] The term " antibody , " as used herein , refers to an immunoglobulin molecule that binds an antigen . Embodiments of an antibody include a monoclonal antibody , polyclonal antibody , human antibody , humanized antibody , chimeric antibody , bispecific or multispecific antibody , or conjugated antibody . The antibodies can be of any class ( e.g. , IgG , IgE , IgM , IgD , IgA ) , and any subclass ( e.g. , IgG1 , IgG2 , IgG3 , IgG4 ) . [ 0043 ] An exemplary antibody of the present disclosure is an immunoglobulin G ( IgG ) type antibody comprised of four polypeptide chains : two heavy chains ( HC ) and two light chains ( LC ) that are cross- linked via inter - chain disulfide bonds . The amino - terminal portion of each of the four polypeptide chains includes a variable region of about 100-125 or more amino acids primarily responsible for antigen recognition . The carboxyl - terminal portion of each of the four polypeptide chains contains a constant region primarily responsible for effector function . Each heavy chain is comprised of a heavy chain variable region ( VH ) and a heavy chain constant region . Each light chain is comprised of a light chain variable region ( VL ) and a light chain constant region . The IgG isotype may be further divided into subclasses ( e.g. , IgG1 , IgG2 , IgG3 , and IgG4 ) . [ 0044 ] The VH and VL regions can be further subdivided into regions of hyper - variability , termed complementarity determining regions ( CDRs ) , interspersed with regions that are more conserved , termed framework regions ( FR ) . The CDRs are exposed on the surface of the protein and are important regions of the antibody for antigen binding specificity . Each VH and VL is composed of three CDRs and four FRs , arranged from amino - terminus to carboxyl - terminus in the following order : FR1 , CDRI , FR2 , CDR2 , FR3 , CDR3 , FR4 . Herein , the three CDRs of the heavy chain are referred to as " HCDR1 , 30574 WO HCDR2 , and HCDR3 " and the three CDRs of the light chain are referred to as " LCDR1 , LCDR2 and LCDR3 " . The CDRs contain most of the residues that form specific interactions with the antigen . Assignment of amino acid residues to the CDRs may be done according to the well - known schemes , including those described in Kabat ( Kabat et al . , " Sequences of Proteins of Immunological Interest , " National Institutes of Health , Bethesda , Md . ( 1991 ) ) , Chothia ( Chothia et al . , " Canonical structures for the hypervariable regions of immunoglobulins " , Journal of Molecular Biology , 196 , 901-9( 1987 ) ; Al - Lazikani et al . , " Standard conformations for the canonical structures of immunoglobulins " , Journal of Molecular Biology , 273 , 927-948 ( 1997 ) ) , North ( North et al . , " A New Clustering of Antibody CDR Loop Conformations " , Journal of Molecular Biology , 406 , 228-256 ( 2011 ) ) , or IMGT ( the international ImMunoGeneTics database available on at www.imgt.org ; see Lefranc et al . , Nucleic Acids Res . 1999 ; 27 : 209-212 ) . [ 0045 ] Embodiments of the present disclosure also include antibody fragments or antigen - binding fragments that , as used herein , comprise at least a portion of an antibody retaining the ability to specifically interact with an antigen or an epitope of the antigen , such as Fab , Fab ' , F ( ab ' ) 2 , Fv fragments , scFv antibody fragments , scFab , disulfide - linked Fvs ( sdFv ) , a Fd fragment . [ 0046 ] The antibodies of the present invention are monoclonal antibodies . Monoclonal antibodies are antibodies derived from a single copy or clone including , for example , any eukaryotic , prokaryotic or phage clone , and not the method by which it is produced . Monoclonal antibodies can be produced , for example , by hybridoma technologies , recombinant technologies , phage display technologies , synthetic technologies , e.g. , CDR - grafting , or combinations of such or other technologies known in the art . [ 0047 ] Methods of producing and purifying antibodies are well known in the art and can be found , for example , in Harlow and Lane ( 1988 ) , Antibodies , A Laboratory Manual , Cold Spring Harbor Laboratory Press , Cold Spring harbor , N.Y. , chapters 5-8 and 15 , ISBN 0-87969-314-2 . For example , mice or rabbits including transgenic mice or rabbits as known in the art , may be immunized with human XCL1 , or portions thereof , and the resulting antibodies can be recovered , screened , purified , and the amino acid sequences determined using conventional methods well known in the art . [ 0048 ] In particular embodiments of the present invention , the antibody , or the nucleic acid encoding same , is provided in isolated form . As used herein , the term " isolated " refers to a protein , peptide , or 30574 WO nucleic acid which is free or substantially free from other macromolecular species found in a cellular environment . [ 0049 ] The antibodies of the present invention may be prepared and purified using known methods . For example , cDNA sequences encoding a HC ( for example the amino acid sequence given by SEQ ID NO : 19 or SEQ ID NO : 20 ) and a LC ( for example , the amino acid sequence given by SEQ ID NO : or SEQ ID NO : 22 ) may be cloned and engineered into a GS ( glutamine synthetase ) expression vector . In one example , an antibody has a heavy chain with amino acid sequence given by SEQ ID NO : 19 and a light chain with amino acid sequence given by SEQ ID NO : 21 ; in such an instance , CDNA sequences which may be used to express the chains are provided by SEQ ID NO : 23 and SEQ ID NO : 24 , respectively . In another example , an antibody has a heavy chain with amino acid sequence given by SEQ ID NO : 20 and a light chain with amino acid sequence given by SEQ ID NO : 22 ; in this instance , cDNA sequences which may be used to express the chains are provided by SEQ ID NO : and SEQ ID NO : 26 , respectively . [ 0050 ] The engineered immunoglobulin expression vector may then be stably transfected into CHO cells . As one of skill in the art will appreciate , mammalian expression of antibodies will result in glycosylation , typically at highly conserved N - glycosylation sites in the Fc region . Stable clones may be verified for expression of an antibody specifically binding to human XCL1 ( for example , as represented by a recombinantly produced peptide comprising SEQ ID NO : 1 ) . Positive clones may be expanded into serum - free culture medium for antibody production in bioreactors . Media , into which an antibody has been secreted , may be purified by conventional techniques . For example , the medium may be conveniently applied to a Protein A or G Sepharose FF column that has been equilibrated with a compatible buffer , such as phosphate buffered saline . The column is washed to remove nonspecific binding components . The bound antibody is eluted , for example , by pH gradient and antibody fractions are detected , such as by SDS - PAGE , and then pooled . The antibody may be concentrated and / or sterile filtered using common techniques . Soluble aggregate and multimers may be effectively removed by common techniques , including size exclusion , hydrophobic interaction , ion exchange , or hydroxyapatite chromatography . The product may be immediately frozen , for example at -70 ° C , or may be lyophilized . 30574 WO [ 0051 ] The antibodies of the present invention can be used in the treatment of patients . More particularly the antibodies of the present invention are expected to be useful in treating immune- mediated diseases or disorders , which include alopecia areata , vitiligo , asthma , RA , MS , UC , psoriasis , and type 1 diabetes . As used interchangeably herein , " treatment " and / or " treating " and / or " treat " are intended to refer to all processes wherein there may be a slowing , interrupting , arresting , controlling , stopping , or reversing of the progression of the disorders described herein , but does not necessarily indicate a total elimination of all disorder symptoms . Treatment includes administration of an antibody , or pharmaceutical composition thereof , of the present invention for treatment of a disease or condition in a human that would benefit from a reduction in XCL1 and / or XCL2 activity , and includes : ( a ) inhibiting further progression of the disease , i.e. , arresting its development ; or ( b ) relieving the disease , i.e. , causing regression of the disease or disorder , or alleviating symptoms or complications thereof . [ 0052 ] As used interchangeably herein , the term " patient , " " subject , " and " individual , " refers to a human . In certain embodiments , the patient is further characterized with a disease , disorder , or condition ( e.g. , an immune - mediated disease ) that would benefit from a reduction in XCL1 and / or XCL2 activity . In other embodiments , the patient is further characterized as being at risk of developing an immune - mediated disease , disorder , or condition that would benefit from a reduction in XCLand / or XCL2 activity . [ 0053 ] As referred to herein , the term " epitope " refers to the amino acid residues , of an antigen , that are bound by an antibody . An epitope can be a linear epitope , a conformational epitope , or a hybrid epitope . [ 0054 ] The term " epitope " may be used in reference to a structural epitope . A structural epitope , according to some embodiments , may be used to describe the region of an antigen which is covered by an antibody ( e.g. , an antibody's footprint when bound to the antigen ) . In some embodiments , a structural epitope may describe the amino acid residues of the antigen that are within a specified proximity ( e.g. , within a specified number of Angstroms ) of an amino acid residue of the antibody . [ 0055 ] The term " epitope " may also be used in reference to a functional epitope . A functional epitope , according to some embodiments , may be used to describe amino acid residues of the antigen that 30574 WO interact with amino acid residues of the antibody in a manner contributing to the binding energy between the antigen and the antibody . [ 0056 ] An epitope can be determined according to different experimental techniques , also called " epitope mapping techniques . " It is understood that the determination of an epitope may vary based on the different epitope mapping techniques used and may also vary with the different experimental conditions used , e.g. , due to the conformational changes or cleavages of the antigen induced by specific experimental conditions . Epitope mapping techniques are known in the art ( e.g. , Rockberg and Nilvebrant , Epitope Mapping Protocols : Methods in Molecular Biology , Humana Press , 3rd ed . 2018 ) , including but not limited to , X - ray crystallography , nuclear magnetic resonance ( NMR ) spectroscopy , site - directed mutagenesis , species swap mutagenesis , alanine - scanning mutagenesis , hydrogen - deuterium exchange ( HDX ) and cross - blocking assays . [ 0057 ] The terms " bind , " " binds , " and " binding " as used herein are intended to mean , unless indicated otherwise , the ability of a protein or molecule to form a chemical bond or attractive interaction with another protein or molecule , which results in proximity of the two proteins or molecules as determined by common methods known in the art . With particular reference to binding of antibody to antigen , the terms " bind , " " binds , " and " binding " refer to the interaction of one or more of the CDRs of the hypervariable region of the antibody with the antigen ; that is , specific binding may be conferred to the antibody by one or more of the CDRs . [ 0058 ] The terms " specifically binds " and " specific binding " as used herein refer to the situation in which one member of a specific binding pair does not significantly bind to molecules other than its specific binding partner ( s ) . For instance , certain antibodies may be said to specifically bind to XCLand XCL2 , but not other similar proteins ( such as chemokines , etc. ) An antibody may be said to bind specifically to an antigen if it binds at least about 50 % greater , or 2 - fold greater , or 20 - fold greater , or - fold greater , or 100 - fold greater than it binds a different antigen or non - antigen target as measured by a technique available in the art , e.g. , competition ELISA or KD measurement with a BIACORE or KINEXA assay . [ 0059 ] An antibody of the present invention can be incorporated into a pharmaceutical composition which can be prepared by methods well known in the art and comprise an antibody of the present invention and one or more pharmaceutically acceptable carrier ( s ) and / or diluent ( s ) ( e.g. , Remington , 30574 WO The Science and Practice of Pharmacy , 22nd Edition , Loyd V. , Ed . , Pharmaceutical Press , 2012 , which provides a compendium of formulation techniques as are generally known to practitioners ) . Suitable carriers for pharmaceutical compositions include any material which , when combined with an antibody of the present invention , retains the molecule's activity and is non - reactive with the patient's immune system . [ 0060 ] A pharmaceutical composition comprising an antibody of the present invention can be administered to a patient at risk for , or exhibiting , diseases or disorders as described herein by parental routes ( e.g. , subcutaneous , intravenous , intraperitoneal , intramuscular , or transdermal ) . A pharmaceutical composition of the present invention contains an " effective " or " therapeutically effective " amount , as used interchangeably herein , of an antibody of the present invention . An effective amount refers to an amount necessary ( at dosages and for periods of time and for the means of administration ) to achieve the desired therapeutic result . An effective amount of an antibody may vary according to factors such as the disease state , age , sex , and weight of the individual , and the ability of the antibody to elicit a desired response in the individual . An effective amount is also one in which any toxic or detrimental effects of the antibody of the present invention are outweighed by the therapeutically beneficial effects . [ 0061 ] Percent homology , as used in the present disclosure , in the context of two or more amino acid sequence refers to two or more sequences having a specified percentage of amino acid residues that are the same , when compared and aligned for maximum correspondence , as measured using a sequence comparison algorithm ( e.g. , BLASTP and BLASTN or other algorithms available to persons of skill ) or by visual inspection . Depending on the application , the percent homology can exist over a region of the sequence being compared , e.g. , over a functional domain , or , alternatively , exist over the full length of the two sequences to be compared . By way of example , percent homology of a sequence may be compared to a reference sequence . For example , when using a sequence comparison algorithm , test and reference sequences may be input into a computer ( and subsequence coordinates may be further designated if desired along with sequence algorithm program parameters ) . The sequence comparison algorithm then calculates the percent sequence identity or homology for the test sequence ( s ) relative to the reference sequence ( s ) , based on the designated program parameters . Exemplary sequence alignment and / or homology algorithms are available through , Smith & 30574 WO Waterman , Adv . Appl . Math . 2 : 482 ( 1981 ) , Needleman & Wunsch , J. Mol . Biol . 48 : 443 ( 1970 ) , Pearson & Lipman , Proc . Natl . Acad . Sci . USA 85 : 2444 ( 1988 ) , GAP , BESTFIT , FASTA , and TFASTA ( in the Wisconsin Genetics Software Package , Genetics Computer Group , 575 Science Dr. , Madison , Wis . ) , or by visual inspection ( see generally Ausubel et al . , infra ) . One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm , which is described in Altschul et al . , J. Mol . Biol . 215 : 403-410 ( 1990 ) . Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information ( www.ncbi.nlm.nih.gov/ ) . [ 0062 ] It is known that in some cases , recombinant production of antibodies can lead to modifications of the antibody . Examples of such modifications include loss of a lysine or glycine at the C - terminus of the HC ; conversion of a glutamine or glutamic acid at the N - terminus of the heavy chain to pyroglutamic acid ; or conversion of a glutamine or glutamic acid at the N - terminus of a lambda light chain to pyroglutamic acid . This may result in production a heterogeneous mix of antibodies , including those with no modification , a single modification , or multiple modifications . The antibodies of the present disclosure may undergo such modifications and are encompassed by the claims derived therefrom . [ 0063 ] In one embodiment , the antibody of the present invention binds XCL1 and neutralizes XCL1 . In one embodiment , the antibody of the present invention binds XCL2 and neutralizes XCL2 . [ 0064 ] In one embodiment , the antibody of the present invention impedes or prevents the migration of a cell expressing XCR1 . In a specific embodiment , the antibody of the present invention neutralizes dendritic cell migration . In one embodiment , the dendritic cell is a cDC1 . [ 0065 ] In one embodiment , the antibody of the present invention has a KD for XCL1 of less than 5.0 . x 10-11 M. In one embodiment , the antibody of the present invention has a KD for XCL2 of less than 1.0 x 10-11 M. [ 0066 ] In one embodiment , the antibody of the present invention binds to XCL1 in its u0000 - sheet monomer configuration . In one embodiment , the antibody of the present invention binds to XCL1 in its dimeric configuration . 30574 WO [ 0067 ] In one embodiment , the antibody of the present invention binds to XCL2 in its u0000 - sheet monomer configuration . In one embodiment , the antibody of the present invention binds to XCL2 in its dimeric configuration . [ 0068 ] In one embodiment , the antibodies disclosed herein bind homodimeric XCL1 . In an embodiment , the antibodies disclosed herein bind homodimeric XCL2 . In one embodiment , the antibodies herein bind heterodimeric complexes which include one monomer of XCL1 and one monomer of XCL2 . In one embodiment , the antibodies disclosed herein bind to free monomeric XCLand / or XCL2 , and / or free homodimeric XCL1 , and / or free homodimeric XCL2 , and / or free heterodimers of XCL1 and XCL2 . In one embodiment , the antibodies of the present disclosure bind to homodimeric XCL1 bound to GAGs , and / or homodimeric XCL2 bound to GAGs , and / or heterodimers of XCL1 and XCL2 bound to GAGs . > > [ 0069 ] In one embodiment , antibody binds a sequence given by SEQ ID NO : 29 ( that is , threonine 16 , glutamine 17 , arginine 18 , leucine 19 , and proline 20 ) . In an embodiment , the antibody binds sequence given by SEQ ID NO : 29 along with lysine 42 and arginine 43. In one embodiment , an antibody of the present invention binds one or more amino acids of SEQ ID NO : 27 ; or two or more amino acids of SEQ ID NO : 27 ; or three or more amino acids of SEQ ID NO : 27. In one embodiment , an antibody of the present invention binds one or more amino acids of SEQ ID NO : 28 ; or two or more amino acids of SEQ ID NO : 28. In an embodiment , an antibody of the present invention binds one or more amino acids of SEQ ID NO : 29. In one embodiment , an antibody of the present invention binds one or more amino acids of SEQ ID NO : 27 and one or more amino acids of SEQ ID NO : 28 ; or two or more amino acids of SEQ ID NO : 27 and two or more amino acids of SEQ ID NO : 28 . [ 0070 ] In one embodiment , an antibody of the present invention has anti - inflammatory activity , and may be used as a therapeutic agent . [ 0071 ] In one aspect , the antibody of the present invention binds to XCL1 and / or XCL2 , but does not specifically bind to another chemokine . 30574 WO EXAMPLES EXAMPLE 1 : Expression of recombinant XCL1 and variants thereof , and XCL[ 0072 ] Each protein ( human XCL1 , SEQ ID NO : 1 ; human XCL2 , SEQ ID NO : 2 ; CC3 , SEQ ID NO : ; and CC5 , SEQ ID NO : 4 ) was expressed as a fusion protein structured with an 8xHis tag at the N- terminus , the well - known BRIL fusion protein , an enterokinase ( EK ) cleavage site , and then the specific protein ( XCL1 , etc. ) at the C - terminal end . A fusion protein expression plasmid was used to create a stable CHO - K1 cell line for each protein by methods known in the art . The fusion proteins were expressed and secreted in chemically defined media . The media was harvested , and fusion proteins were captured on a Ni - IMAC column ( for example , as is available from Pierce ) and eluted with 375 mM imidazole at pH 7.5 . The Ni - IMAC pool was dialyzed into the EK digestion buffer ( mM Tris , 100 mM NaCl , 5 mM CaCl2 ; pH 8 ) , and the fusion protein was incubated overnight with EK . The free protein was captured with Capto - S resin ( Cytiva Cat # 17544101 ) before polishing by size exclusion chromatography ( Superdex 75 - Cytiva Cat # 28989333 ) . Protein identity was confirmed by mass spectrometric analysis .

EXAMPLE 2 : Expression of anti - XCL1 antibodies – [ 0073 ] Exemplified anti - XCL1 antibodies of the present invention are presented in Table 1 below . Antibodies were produced in stable CHO - K1 cell lines in a chemically defined media . They were captured on PRISMA resin ( Cytiva - Cat # 1754981 ) and eluted with a pH 3 buffer . PRISMA pool was brought to pH 5 to precipitate host cell proteins ( HCP ) . The proteins were polished by anion exchange chromatography ( AEX ) using a POROS 50 HS ( Thermo Fisher Cat # 1335906 ) and then dialyzed into 10 mM Histidine , 275 mM Sucrose ; pH 6. Material was analyzed by analytical size exclusion chromatography ( SEC ) and confirmed using mass spectrometry .

Table 1 : SEQ ID Nos of Exemplified Antibody Amino Acid Sequences HC HCVR H CDRH CDRH CDRLC LCVR L CDRL CDRL CDR3 30574 WO Ab 19 15 10 11 13 21 17 Ab 20 16 10 11 14 22 18 ∞ [ 0074 ] A control antibody specifically binding to monomeric XCL1 ( that is , which binds CC3 with high affinity and does not bind CC5 ; see below Examples ) , with HC amino acid sequence given by SEQ ID NO : 27 and LC amino acid sequence given by SEQ ID NO : 28. cDNA sequences which may be used for expressing the HC and LC are given by SEQ ID NO : 29 and SEQ ID NO : 30 , respectively . This antibody is referred to herein as monomer - binding control ( MBC ) . [ 0075 ] The relationship of the various regions of exemplified anti - XCLI antibodies is as follows ( numbering of amino acids applies linear numbering ; assignment of amino acids to variable domains is based on the International Immunogenetics Information ®metsyS available at www.ingt.org . ) Assignment of amino acids to CDR domains for each antibody exemplified is based on the well - known North numbering convention , with the exception of HCDR2 , which C - terminus is based on the well- known Kabat numbering convention . [ 0076 ] Exemplified antibodies of the present disclosure are identified as possessing high binding affinity and being chemically and physically stabile including aggregation and solubility consistent with therapeutic parental administration . The exemplified antibodies of the present disclosure are also identified as possessing low immunogenicity ( including low nonspecific and / or serum binding ) and possessing pharmacokinetic properties consistent with therapeutic parental administration for the treatment of immune - mediated diseases . In one aspect , the antibodies disclosed herein are human antibodies of the IgG1 isotype . The antibodies disclosed herein are monoclonal antibodies . Antibodies of other isotypes ( for example , IgG4 , and so forth ) are also contemplated by this disclosure .

EXAMPLE 3 : Properties of XCL1 variants CC3 and CC [ 0077 ] XCL1 is a metamorphic protein that readily undergoes major conformational rearrangements depending on salt and temperature conditions . High salt and low temperature ( 10 ° C ) favors an active 30574 WO monomer conformation having an α - helix motif typical of canonical chemokine folds that can signal through the cognate receptor , XCR1 , while low salt and higher temperature ( 40 ° C ) favors an inactive u0000 - sheet configuration which can dimerize and bind to endogenous glycosaminoglycans ( GAGs ) . [ 0078 ] To study the properties of each configuration separately , engineered variant proteins were expressed and purified . CC3 ( SEQ ID NO : 3 ) is an engineered variant of XCL1 stabilized in the active , α - helix - containing monomer conformation through creation of a disulfide bond when residues and 59 are mutated to cysteine ( V21C / V59C ) . Likewise , mutations to cysteine at residues 36 and ( A36C / A49C ) create an alternative disulfide structure that stabilizes the inactive u0000 - sheet configuration which tends toward the dimeric state , termed CC5 ( SEQ ID NO : 4 ) . These variants are known in the art and were prepared as specified in the literature ( see Biopolymers 20Oct ; 112 ( 10 ) : e23402 doi : 10.1002 / bip.23402 . Epub 2020 Sep 28. ) [ 0079 ] XCR1 is expressed on several types of dendritic cells ( DC ) , primarily type 1 conventional dendritic cells ( cDC1 ) . cDC1 are antigen presenting cells ( APC ) adept at antigen presentation to CD8 + T cells in the context of major histocompatibility complex I. As CD8 + T cells secrete XCL1 , a chemotactic gradient is generated , attracting XCR1 - bearing cells to the site of the CD8 + T cells ( for instance , within a tumor or autoimmune site ) , augmenting the immune response . See Dorner et al . , Immunity 31 ( 2009 ) 823-833 ; Audsley et al . Cells 9 ( 2020 ) 565 1-22 . [ 0080 ] Recombinant human XCL1 , CC3 , and CC5 were prepared and evaluated in a migration assay utilizing a murine Ba / F3 cell line stably overexpressing human XCR1 . This cell line migrates toward an XCL1 chemokine gradient in vitro , in a 5 Mµ Boyden Chamber assay plate . The assay is set up with 150,000 cells / well in the top chamber with various concentrations of XCL1 added to the bottom chamber . Cells are permitted to migrate over approximately 4 hours at 37 ° C , with the translocated cells quantitated using a luminescent cell viability readout . Results are shown below in Table 2 . 30574 WO TABLE 2 : Migration of XCR1 - expressing cells in the presence of XCL1 variants Ligand GeoMean EC50 ( nM ) Error ( delta method ) Human XCLwild - type 858.4 82.0 Human XCLCC3 114.7 65.

Human XCLCC5 Inactive [ 0081 ] For CC3 , robust migration occurs with an EC50 around 114.7 nM compared to wild - type XCL( 858.4 nM ) . For CC5 , migration was not observed . These data confirm observations in the literature indicating that recombinant CC3 and CC5 represent the two distinct conformations of XCL1 .

EXAMPLE 4 : Binding Kinetics and Affinity [ 0082 ] The affinity and binding kinetics of antibodies of the present invention and the MBC to human XCL1 , human XCL2 , mouse XCL1 , cynomolgus ( cyno ) XCL1 , and rat XCL1 were determined by surface plasmon resonance using Biacore 8K ( GE Healthcare ) . The procedure generally follows the " Instrument Handbook . " Antibody was first captured on a Biacore Protein A chip followed by flowing a serial concentration of antigens from 400 nM down to 0.039 nM in 2 - fold serial dilution in PBS- P20 - BSA ( 0.1 % surfactant P20 , 0.1 mg / mL BSA ) . All measurements are carried out at 25 ° C . [ 0083 ] The multi - cycle kinetics setting that runs each analyte concentration in a separate cycle regenerating the surface after each sample injection is used . The regeneration is optimized to maintain consistent surface properties from cycle to cycle . Each cycle starts with a 3 minute injection of antibody ( approximately 150 response units ( RU ) capture level ) at 10 lµ / min flow rate , followed by minute injection of antigen at 50 lμ / min flow rate and a 15 min dissociation phase in PBS - P20 - BSA . The chip surface was then regenerated with 30 seconds injection of pH 1.5 , 10 mM glycine buffer at lμ / min flow rate for three times . The data was fit to a 1 : 1 binding model to derive ka and kd , and to calculate Kd . 30574 WO [ 0084 ] Following procedures essentially as described above , the following parameters are obtained for the antibodies in the present invention .

TABLE 3 : Antibody affinity and kinetics with various XCL1 antigens huXCL1 msXCL1 cynoXCL1 Rat XCLAntibody ka ( 1 / Ms ) kd ( 1 / s ) Kd ( M ) ka ( 1 / Ms ) kd ( 1/5 ) Kd ( M ) ka ( 1 / Ms ) kd ( 1/5 ) Kd ( M ) ka ( 1 / Ms ) kd ( 1 / s ) Kd ( M ) 5.0E + 07 6.7E - 04 1.3E - 11 1.1E + 06 1.4E - 04 1.3E - 10 1.5E + 08 5.7E - 3.9E- 9.6E + 06 6.1E - 6.3E- 9.2E + 06 2.9E - 04 3.2E - 11 6.1E + 05 2.1E - 04 3.5E - 10 2.5E + 07 2.8E - 1.1E- 4.5E + 06 1.5E - 3.2E- MBC 3.5E + 07 8.2E - 05 2.4E - 12 1.9E + 05 1.3E - 04 6.8E - 10 2.2E + 07 12.9E - 1.3E- 4.0E + 06 < 1e - 6 < 1e - huXCL2 CC3 CCAntibody ka ( 1 / Ms ) kd ( 1 / s ) Kd ( M ) ka ( 1 / Ms ) 1.8E + 07 6.4E - 05 3.5E - kd ( 1 / s ) Kd ( M ) ka ( 1 / Ms ) | kd ( 1 / s ) | Kd ( M ) No Binding 8.3E + 06 2.1E - 04 2.6E - 2 6.8E + 06 5.2E - 05 7.7E - 12 No Binding 8.2E + 06 1.8E - 04 2.2E - MBC 2.9E + 06 1.0E - 05 3.5E - 12 1.1E + 05 2.1E - 04 1.9E - No Binding [ 0085 ] Notably , Antibodies 1 and 2 bind CC5 with good affinity but not CC3 , whereas the opposite is observed in the monomer - specific binding of MBC . This may be because the introduction of cysteines in CC3 may interfere with the epitope to which Antibodies 1 and 2 bind . Crystallography experiments support Antibody 1 and Antibody 2 binding to XCL1 in u0000 - sheet monomer form . In one aspect , Antibodies 1 and 2 can be referred to as primarily and preferentially binding the dimeric form , and these results demonstrate that Antibodies 1 and 2 can bind to homodimeric XCL1 . 30574 WO EXAMPLE 5 : Neutralization of XCL1 and XCL2 : human XCR1 migration assay [ 0086 ] XCL1 and XCL2 are highly conserved chemokines with chemotactic activity mediated through the XCR1 receptor . The murine cell line Ba / F3 was stably transfected with human XCR1 receptor to assess the ability of XCL1 / 2 to induce migration in an in vitro system . [ 0087 ] For this assay , 150,000 recombinant hXCR1 / Ba / F3 cells were seeded in the top chamber of a Mμ5 Boyden Chamber assay plate with XCL1 concentrations empirically determined for each species added to the bottom chamber . The cells were allowed to migrate toward the XCL1 / 2 gradient for approximately 4 hours with the cells in the bottom chamber assayed by luminescent cell viability readout . In this assay , human XCL1 , human XCL2 , mouse , rat and cyno XCL1 all induce migration . After a dose - response of ligand was conducted , a single concentration of XCL1 / 2 was selected and used into the bottom chamber of the Boyden Chamber plate along with a dose - response of each mAB tested . Neutralization IC50 values for each antibody were determined . As seen in Table 4 below , all antibodies were able to neutralize human XCL1 and XCL2 , as well as rat , mouse and cyno XCL1 . TABLE 4 : Neutralization of XCL1 and XCL2 by antibodies of the present invention Ligand ( XCL1 if not Ligand GeoMean IC50 ( nM ) Error ( delta specified ) ( nM ) Antibody method ) n Log Mean N sd Day Rat 7.79 2.56 0.25 0.41 0.07 Rat 7.79 2 5.65 0.29 3 0.75 0.04 Mouse 0.49 1 1.07 0.3 0.03 0.15 Mouse 0.49 2 ﻞﻫ 2.98 0.90 3 0.47 0.23 Cyno 7.79 4.07 0.22 3 0.61 0.04 Cyno 7.79 2 4.51 0.16 3 0.65 0.03 human XCL2 7.79 2.30 0.01 3 0.36 0.00 human XCL2 7.79 2 2.53 0.21 3 0.40 0.06 human XCL1 2.43 0.60 0.05 3 -0.22 0.06 human XCL1 2.43 2 0.66 0.10 3 -0.18 0.12 3 30574 WO [ 0088 ] The results provided in Tables 3 and 4 demonstrate that both exemplified antibodies bind both XCL1 and XCL2 with high affinity , and neutralize both chemokines in in vitro assays . Therefore it will be appreciated that the antibodies of the present disclosure are useful for binding and neutralization of XCL1 , XCL2 , or both , including in a therapeutic context . It will be appreciated that the antibodies as disclosed herein bind homodimeric XCL1 , and that because of the sequence similarity between XCL1 and XCL2 , that the antibodies bind homodimeric XCL2 . The antibodies will also bind heterodimeric complexes which may form and which include one monomer of XCL1 and XCL2 . The antibodies of the present disclosure may bind to free monomeric XCL1 or XCL2 , or free homodimeric XCL1 , or free homodimeric XCL2 , or free heterodimers of XCL1 and XCL2 . The antibodies of the present disclosure may also bind to homodimeric XCL1 bound to GAGs , or homodimeric XCL2 bound to GAGs , or heterodimers of XCL1 and XCL2 bound to GAGs .

EXAMPLE 6 : Neutralization of XCL1 activity by antibodies : dendritic cell migration [ 0089 ] XCL1 and XCL2 stimulate migration of XCR1 expressing conventional type 1 dendritic cells ( CDC1 ) . To demonstrate XCL1 / 2 antibody neutralization activity , human cDC1 cells were differentiated from CD34 + stem cells and their migration towards XCL1 through a Boyden Chamber was measured by flow cytometry . Briefly , CD34 + cells from human cord blood were differentiated over 3 weeks in a cytokine cocktail on a feeder layer expressing DLL1 ( adapted from Balan et al . doi : 10.1016 / j.celrep.2018.07.033 ) . Cells were enriched using a myeloid dendritic cell kit ( StemCell ; lineage negative , HLA - DR + , CD11c ) . 10 ng / mL XCL1 + antibody dose titrations in triplicate were pre - incubated 1 hour and added to the bottom chamber of the Boyden Chamber . Antibodies employed were Antibody 1 , Antibody 2 , and hIgG1 - AAS isotype control antibody . 150,000 enriched , differentiated cells were added to the top chamber of the Boyden Chamber and incubated for 2 hours . Migrated human cDC1 cells in the bottom chamber were stained ( live / dead , CLEC9A + , XCR1 + ) and counted . The present XCL1 / 2 antibodies completely inhibited cDC1 migration towards XCL1 across replicate experiments and the GeoMean of IC50 values ( nM ) and associated error ( delta method ) are shown in Table 5 . 30574 WO TABLE 5 : Potency of XCL1 / 2 antibody inhibition of XCL1 stimulated primary human cDCmigration Antibody Control GeoMean IC50 ( nM ) Error ( delta method ) 0.194 0.0 0.363 0.0 No inhibition -- EXAMPLE 7 : Anti - inflammatory effects of antibody in mouse [ 0090 ] The immunosuppressive activities of Antibodies 1 and 2 were assessed in a mouse model of oxazolone - induced delayed - type hypersensitivity on mouse ears and compared to an anti - CDdepleting antibody ( Bio X Cell # BE0117 ) . Mice were treated with anti - XCL1 / 2 antibodies on day 0 , approximately 1 hour prior to sensitization with 3 % oxazolone in acetone on their shaved , ventral - side surface . On day 6 , mice were challenged with 2 % oxazolone administration on the ear with change in ear swelling measured on day 7. Results are shown in Table 6 , as delta ear thickness with anti - XCL1 / antibodies compared to a matched isotype .

TABLE 6 : Anti - inflammatory action in mouse model Antibody GeoMean Error n ( delta thickness ( mm ) compared to isotype ( % ) ) -40 3.5 2 -43 1.0 Anti - CD8 -38 4.2 30574 WO EXAMPLE 8 : Epitope mapping and determination [ 0091 ] Binding sites for the exemplified antibodies on the surface of XCL1 were determined by HDX and X - ray crystallography , techniques well known in the art . By HDX , Antibodies 1 and 2 were found to bind a non - linear epitope formed of residues within residues 12 to 20 ( SEQ ID NO : 27 ) , and within residues 35 to 46 , SEQ ID NO : 28. An X - ray crystal structure of a fragment antigen - binding region ( Fab ) complexed with human XCL1 , to 2.77 angstrom resolution , further suggests that the specific residues involved in binding are those whose sequence is given by SEQ ID NO : 29 ( that is , threonine , glutamine 17 , arginine 18 , leucine 19 , and proline 20 ) , along with lysine 42 and arginine 43. An antibody of the present invention may therefore bind one or more amino acids of SEQ ID NO : 27 ; or two or more amino acids of SEQ ID NO : 27 ; or three or more amino acids of SEQ ID NO : 27. An antibody of the present invention may bind one or more amino acids of SEQ ID NO : 28 ; or two or more amino acids of SEQ ID NO : 28. An antibody of the present invention may bind one or more amino acids of SEQ ID NO : 29. An antibody of the present invention may bind one or more amino acids of SEQ ID NO : 27 and one or more amino acids of SEQ ID NO : 28 ; or two or more amino acids of SEQ ID NO : 27 and two or more amino acids of SEQ ID NO : 28 . [ 0092 ] The antibodies exemplified herein bind to a non - linear epitope of XCL1 / XCL2 . In one instance , the antibodies bind to a monomeric u0000 - sheet configuration of XCL1 or XCL2 . In another instance , the antibodies bind to a dimeric form of XCL1 or XCL2 . In such a case , the antibody may bind to a single one of the two monomer subunits of chemokine which come together to define the dimer . For example , the antibody may bind to amino acid residues defined by SEQ ID NO : 29 and lysine 42 and / or arginine 43 of a first monomeric unit of XCL1 . This disclosure likewise contemplates an antibody which binds dimeric XCL1 or XCL2 by binding a non - linear epitope which includes residues of both monomers of the dimer . Another binding mode contemplated herein is that of binding monomeric u0000 - sheet XCL1 or XCL2 in such a way that dimerization is prevented .

EXAMPLE 9 : GAG - binding assay [ 0093 ] As mentioned , the u0000 - sheet configuration of XCL1 / 2 is able to dimerize and cannot bind the XCR1 receptor . Instead , in this form XCL1 / 2 binds to cell surface GAGs , which is thought to help establish a chemoattractive gradient . Antibodies 1 and 2 bind to the u0000 - sheet configuration , 30574 WO preferentially to the dimer form , and their ability to bind to XCL1 / 2 dimers bound to cell surface GAGs using flow cytometry was studied . [ 0094 ] 293T cells were incubated with 1 gµ / mL of various preparations of XCL1 or XCL2 , and 1nM of either Antibody 1 , Antibody 2 , or hIgG1 - AAS isotype control antibody ( " " Control " ) for 1 hour . This was followed by staining with anti - human - Fc / PE secondary antibody ( Life Technologies 12- 4998-82 ) to detect the antibody - XCL1 / 2 complex bound to cell surface GAG . In another condition , the binding of antibody - XCL1 / 2 complex is done in the presence of 100 ug / mL heparin to demonstrate the specificity of GAG binding . The mean fluorescence intensity ( MFI ) was measured by flow cytometry for each condition . The experiment was repeated 3 times and the average MFI +/- standard deviation across experiments is shown below in Table 7 .

TABLE 7 : Detection of XCL1 / 2 antibody binding to 293T cell surface XCL1 / 2 - GAG by flow cytometry ( MFI ) Human XCLCyno XCLRat XCL Antibody MFI +/- MFI +/- MFI +/- 1 213.7 136.1 95.3 27.0 112.1 19. 1 + 25.9 9.8 13.1 2.2 18.8 1. heparin 142.7 74.3 777.9 152.0 266.3 35. 2 + 55.8 23.9 24.5 8.8 26.0 6.heparin Control 7.6 0.4 8.4 0.2 8.2 0.

Human XCL2 Mouse XCL Antibody MFI +/- MFI +/- 30574 WO 1 148.9 70.6 49.0 19. 1+ 21.8 6.3 9.2 0. heparin 767.7 289.6 61.3 12. 2 + 57.3 26.9 12.8 0. heparin Control 8.6 0 8.0 0. [ 0095 ] The results above demonstrate that Antibodies 1 and 2 bind to inactive or dimeric XCL1 at the cell surface , and taken together with the results of Example 6 , demonstrate that Antibodies 1 and may bind inactive XCL1 , sequestering it so that it does not activate XCR1 .

EXAMPLE 10 : XCL1 SPECIFICITY OF EXEMPLIFIED ANTIBODIES [ 0096 ] A human serum binding assay was used to identify non - specific targets of Antibodies 1 and 2 , and MBC . Each antibody was coated in a microwell followed by treatment with serum . After an overnight incubation , unbound serum proteins were washed away prior to elution of antibody - bound proteins . These samples were digested with trypsin with peptides identified by mass spectrometry . [ 0097 ] Antibodies 1 and 2 did not bind to other chemokines , but MBC was found to bind a substantial amount of a second , off - target chemokine . Subsequent experiments demonstrated that MBC lost biological activity in the presence of high concentrations of this chemokine ( that is , similar to that expected in the physiological context ) whereas the other Antibodies 1 and 2 were unaffected . 30574 WO SEQUENCE LISTING SEQ ID NO : 1 human XCL VGSEVSDKRTCVSLTTQRLPVSRIKTYTITEGSLRAVIFITKRGLKVCADPQATWVRDVVRSMDRKSNTRNNM IQTKPTGTQQSTNTAVTLTG SEQ ID NO : 2 human XCL VGSEVSHRRTCVSLTTQRLPVSRIKTYTITEGSLRAVIFITKRGLKVCADPQATWVRDVVRSMDRKSNTRNNM IQTKPTGTQQSTNTAVTLTG SEQ ID NO : 3 CCMNNPTNSKRDMØRVCDRVWTAQPDACVKLGRKTIFIVARLSGETITYTKIRSCPLRQTTLSVCTRKDSVESGV IQTKPTGTOOSINTAVTLTG SEQ ID NO : 4 CCVGSEVSDKRTCVSLTTQRLPVSRIKTYTITEGSLROVIFITKRGLKVCCDPQATWVRDVVRSMDRKSNIRNNM IQTKPTGTOOSTNTAVTLTG SEQ ID NO : 5 LCDR1 generic formula for Antibodies 1 and QASQXISDYLN , wherein X is selected from S and D SEQ ID NO : 6 Antibody 1 Light Chain CDR QASQDISDYLN SEQ ID NO : 7 Antibody 2 Light Chain CDR QASQSISDYLN 30574 WO SEQ ID NO : 8 Light Chain CDR2 for Antibodies 1 and YDASAKQT SEQ ID NO : 9 Light Chain CDR3 for Antibodies 1 and QHYDNLGYYT SEQ ID NO : 10 Heavy Chain CDR1 for Antibodies 1 and KASGYTFTSYYMV SEQ ID NO : 11 Heavy Chain CDR2 for Antibodies 1 and VIDPSKGSTNYAQKFQG SEQ ID NO : 12 HCDR3 CDR2 for Antibodies 1 and AREGLZLLAFDI , wherein Z is selected from Q and E SEQ ID NO : 13 Antibody 1 Heavy Chain CDR AREGLELLAFDI SEQ ID NO : 14 Antibody 2 Heavy Chain CDR AREGLOLLAFDI SEQ ID NO : 15 Antibody 1 HCVR 30574 WO X , VOLVQSGAEVKKPGASVKVSCKASGYTFTSYYMVWVRQAPGQGLEWMGVIDPSKGSTNYAQKFQGRVTMTR DISTSTVYMELSSLRSEDTAVYYCAREGLELLAFDIWGQGTMVTVSS , wherein X is Q or pyroglutamic acid SEQ ID NO : 16 Antibody 2 HCVR X VOLVQSGAEVKKPGASVKVSCKASGYTETSYYMVWVRQAFGQGLEWMGVIDPSKGSTNYAQKFQGRVTMTR DTSTSTVYMELSSLRSEDTAVYYCAREGLOLLAFDIWGQGTMVTVSS , wherein X is Q or pyroglutamic acid SEQ ID NO : 17 Antibody 1 LCVR DIQMTQSPSSLSASVGDRVTITCQASQDISDYLNWYQQKPGKAPKLLIYDASAKQTGVPSRFSGSGSGTDFTF TISSLOPEDIATYYCQHYDNLGYYTFGQGTKLEIK SEQ ID NO : 18 Antibody 2 LCVR DIQMTQSPSSLSASVGDRVTITCQASQSISDYLNWYQQKPGKAPKLLIYDASAKQTGVPSRFSGSGSGTDFTF TISSLOPEDIATYYCQHYDNLGYYTFGQGTKLEIK SEQ ID NO : 19 Antibody 1 HC X1VQLVQSGAEVKKPGASVKVSCKASGYTETSYYMVWVRQAPGQGLEWMGVIDPSKGSTNYAQKFQGRVTMTR DTSTSTVYMELSSLRSEDTAVYYCAREGLELLAFDIWGQGTMVTVS SASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTF PAVLOSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKRVE PKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTIMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEY KOKVSNKALPAPIEKTISKAKGQPREPOVYTLPPSREEMTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNH YTQKSLSLSPX2X3 , wherein ₁X is Q or pyroglutamic acid ; X2 is G or absent ; and X3 is K or absent 30574 WO SEQ ID NO : 20 Antibody 2 HC X1VQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMVWVRQAPGQGLEWMGVIDPSKGSTNYAQKFQGRVTMTR DTSTSTVYMELSSLRSEDTAVYYCAREGLOLLAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVE PKSCDKTHTCPPCPAPEAAGGPSVFLEPPKPKOTIMISRTPEVTCVVVSVSHEDPEVKENWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVL HQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPOVYTLPPSREEMT KNQ VSLTCLVKGPYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWOOGNVFSCSVMHEALHNH YTQKSLSLSPX2X3 , wherein ₁X is Q or pyroglutamic acid ; X2 is G or absent ; and X3 is K or absent SEQ ID NO : 21 Antibody 1 LC DIQMTQSPSSLSASVGDRVTITCQASQDISDYLNWYQQKPGKAPKLLIYDASAKQTGVPSRFSGSGSGTDFTF TISSLOPEDIATYYCQHYDNLGYYTFGQGT KLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLINNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO : 22 Antibody 2 LC DIQMTQSPSSLSASVGDRVTITCQASQSISDYLNWYQQK PGKAP KLLIYDASAKQTGVPS RFSGSGSGTDFTF TISSLQPEDIATYYCQHYDNLGYYTFGOGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKOSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS FNRGEC SEQ ID NO : 23 Antibody 1 heavy chain DNA sequence CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCGTGGGGCCTCAGTGAAGGTTTCCTGCAAAGOGT CTGGATACACCTTCACCAGCTATTATATGGTCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGG AGTAATCGACCCTAGTAAAGGTAGCACAAACTACGCACAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGAC ACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCOTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGA GAGAAGGACTGGAACTACTTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCAGCCTCCAC CAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGC CTGGTCAAGGACTACTICO CGGTGACGGTGTCGTGGAACTCAGGCGCGCTGACCAGCGGCGTGCACA CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTT GGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCC 30574 WO PAATCTTGTGACAAAACTCACACATGC TCTTCCC CCGTCAGTCTTCC CCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGTCCGT GAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAG CCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGA ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCA CAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCAT CCATCGAGAAAACCATCTCCAAAGC CCGGGAGGAGATGACCAAGAACCAAGTO AGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATICCAAGCTCACCGT GGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTAC ACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO : 24 Antibody 1 light chain DNA sequence GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGG CGAGTCAGGACATAAGCGACTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTA CGATGCTTCCGCTAAACAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTITC ACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAACATTATGATAATCTCGGATACTACA CTTTTGGCCAAGGGACCAAGCTGGAGATCAAAagaactgtggcggcgccatctgtcttcatcttcccgccato tgatgagcagttgaaatccggaactgcctctgttgtgtgcctgctgaataacttctatcocagagaggccaaa gtacagtggaaggtggataacgccctccaatcgggtaactoccaggagagtgtcacagagcaggacagcaagg acagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgoctg cgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgc SEQ ID NO : 25 Antibody 2 heavy chain DNA sequence CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAAGCGT CTGGATACACCTTCACCAGCTATTATATGGTCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGG AGTAATCGACCCTAGTAAAGGTAGCACAAACTACGCACAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGAC ACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTGCGA GAGAAGGACTGCAACTACTTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCAGCCTCCAC CAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGC CTGGTCAAGGACTACTICO CGGTGACGGTGTCGTGGAACTCAGGCGCGCTGACCAGCGGCGTGCACA CCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTT GGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCC 30574 WO PAATCTTGTGACAAAACTCACACATGC CCGTCAGTCTTCC TCTTCCC CCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGTCCGT GAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAG CCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTGGCTGA ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAC CCATCGAGAAAACCATCTCCAAAGC CAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATO CCGGGAGGAGATGACCAAGAACCAAGTO AGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATICCAAGCTCACCGT GGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTAC ACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO : 26 Antibody 2 light chain DNA sequence GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGG CGAGTCAGAGCATAAGCGACTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTA CGATGCTTCCGCTAAACAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTC ACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAACATTATGATAATCTCGGATACTACA CTTTTGGCCAAGGGACCAAGCTGGAGATCAAAAGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATC TGATGAGCAGTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAA GTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGG ACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTG CGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC SEQ ID NO : 27 XCL1 Epitope 1 ( Antibodies 1 and 2 ) VSLTTORLE SEQ ID NO : 28 XCL1 Epitope 2 ( Antibodies 1 and 2 ) RAVIFITKRGLK SEQ ID NO : 29 XCL1 Epitope 1 subsequence ( Antibodies 1 and 2 ) TQRLP 30574 WO SEQ ID NO : 30 Antibody 1 HCVR without variables QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMVWVRQAPGQGLEWMGVIDPSKGSTNYAQKFQGRVTMTRD TSTSTVYMELSSLRSEDTAVYY CAREGLELLAFDIWGQGTMVTVSS SEQ ID NO : 31 Antibody 2 HCVR without variables QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMVWVRQAPGQGLEWMGVIDPSKGSTNYAQKFQGRVTMTRD TSTSTVYMELSSLRSEDTAVYYCAREGLOLLAFDIWGQGTMVTVSS SEQ ID NO : 32 Antibody 1 HC without variables QVOLVOSGAEVKKPGASVKVSCKASGYTFTSY YM VW VR QAPGOGLEWMGVIDPSKGSTNYAQK FOGRVIMIRD TSTSTVYMELSSLRSEDTAVYYCAREGLELLAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGO LVKDYFPEPVTVSWNSGALTSGVHTFPAVLOSSGLYSLSSVVTVPSSSLGTOTYICNVNHKPSNT KVDKRVEP KSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTPEVIC SVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PRE PQVY TLPPSREEMT KN QV SLTCLVKGEY PSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALANHY TOKSLSLSPGK SEQ ID NO : 33 Antibody 2 HC without variables QVQLVQSGAEVKKPGASVKVSCKASGYTFTS MVWVRQAPGOGLEWMGVIDPSKGSTNYAQKFQGRVTMTRD TSTSTVYMELSSLRSEDTAVYYCAREGLOLLAFDIWGOGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGO LVKDYFPEPVTVSWNSGALTSGVHTFPAVLOSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEP KSCDKTHTCPPCPAPEAAGGPSVFL FPPKPKDTLMISRTPEVICVVVSVSHEDPEVKENWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSREEMT KNQV SLTCLVKGFYPSDIAVEWESNGOPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHY TOKSLSLSPGK

Claims (41)

- 33 - CLAIMS We claim :

1. An antibody that binds human XCL1 comprising a heavy chain variable region ( HCVR ) and a light chain variable region ( LCVR ) , wherein the HCVR comprises complementarity determining regions ( CDRs ) HCDR1 , HCDR2 , and HCDR3 and the LCVR comprises CDRs LCDR1 , LCDR2 , and LCDR3 , wherein the HCDR1 comprises SEQ ID NO : 10 , the HCDR2 comprises SEQ ID NO : 11 , the HCDR3 comprises SEQ ID NO : 12 , wherein Z is selected from Q and E ; the LCDR1 comprises SEQ ID NO : 5 , wherein X is selected from S and D , the LCDR2 comprises SEQ ID NO : 8 , and the LCDR3 comprises SEQ ID NO : 9 .

2. The antibody of claim 1 , wherein HCDR1 comprises SEQ ID NO : 10 ; HCDR2 comprises SEQ ID NO : 11 ; HCDR3 comprises SEQ ID NO : 13 ; LCDR1 comprises SEQ ID NO : 6 ; LCDRcomprises SEQ ID NO : 8 ; and LCDR3 comprises SEQ ID NO : 9 .

3. The antibody of claim 1 or 2 , wherein the LCVR comprises SEQ ID NO : 17 and the HCVR comprises SEQ ID NO : 15 , wherein Xa of SEQ ID NO : 15 is selected from Q and pyroglutamic acid .

4. The antibody of claim 3 , wherein Xa is Q.

5. The antibody of any one of claims 1-4 , wherein the antibody comprises a heavy chain ( HC ) and a light chain ( LC ) , wherein the HC comprises SEQ ID NO : 19 , wherein ₁X of SEQ ID NO : 19 is selected from Q and pyroglutamic acid ; X2 of SEQ ID NO : 19 is G or absent ; and X3 of SEQ ID NO : 19 is K or absent ; and the LC comprises SEQ ID NO : 21 .

6. The antibody of claim 5 , wherein ₁X is Q ; X2 is G ; and X3 is K. - 34 -

7. The antibody of claim 1 , wherein HCDR1 comprises SEQ ID NO : 10 ; HCDR2 comprises SEQ ID NO : 11 ; HCDR3 comprises SEQ ID NO : 14 ; LCDR1 comprises SEQ ID NO : 7 ; LCDRcomprises SEQ ID NO : 8 ; and LCDR3 comprises SEQ ID NO : 9 .

8. The antibody of claim 1 or 7 , wherein the LCVR comprises SEQ ID NO : 18 and the HCVR comprises SEQ ID NO : 16 , wherein Xa of SEQ ID NO : 16 is selected from Q and pyroglutamic acid .

9. The antibody of claim 8 , wherein Xa is Q.

10. The antibody of any one of claims 1 , 7 , and 8 , wherein the antibody comprises a heavy chain ( HC ) and a light chain ( LC ) , wherein the HC comprises SEQ ID NO : 20 , wherein X1 of SEQ ID NO : 20 is selected from Q and pyroglutamic acid ; X2 of SEQ ID NO : 20 is G or absent ; and X3 of SEQ ID NO : 20 is K or absent ; and the LC comprises SEQ ID NO : 22 .

11. The antibody of claim 10 , wherein ₁X is Q ; X2 is G ; and X3 is K.

12. The antibody of any one of claims 1-11 , wherein the antibody binds human XCL2 , dimeric XCL1 , or dimeric XCL2 .

13. The antibody of any one of claims 1-12 , wherein the antibody binds XCL1 in a u0000 - sheet configuration .

14. The antibody of any one of claims 1-13 , wherein the homodimeric XCL1 is bound to glycosaminoglycan on a surface of a cell .

15. The antibody of any one of claims 1-14 , wherein the antibody binds at least one amino acid residue on each monomeric unit of dimeric XCL2 .

16. The antibody of any one of claims 1-15 , wherein the homodimeric XCL2 is bound to glycosaminoglycan on a surface of a cell . - 35 -

17. The antibody of any one of claims 1-16 , which binds a heterodimeric complex comprising one monomer of XCL1 and one monomer of XCL2 .

18. The antibody of any one of claims 1-17 , wherein the antibody binds at least one amino acid of the XCL1 monomer and at least one amino acid of the XCL2 monomer of the heterodimeric complex .

19. The antibody of any one of claims 1-18 , wherein the heterodimeric complex is bound to glycosaminoglycan on a surface of a cell .

20. An antibody that binds human XCL1 comprising a heavy chain variable region ( HCVR ) having at least 95 % sequence identity to SEQ ID NO : 15 and a light chain variable region ( LCVR ) having at least 95 % sequence identity to SEQ ID NO : 17 .

21. The antibody of claim 20 , comprising a heavy chain ( HC ) having at least 95 % sequence identity to SEQ ID NO : 19 , and a light chain ( LC ) having at least 95 % sequence identity to SEQ ID NO : .

22. An antibody that binds human XCL1 comprising a heavy chain variable region ( HCVR ) having at least 95 % sequence identity to SEQ ID NO : 16 and a light chain variable region ( LCVR ) having at least 95 % sequence identity to SEQ ID NO : 18 .

23. The antibody of claim 22 , comprising a heavy chain ( HC ) having at least 95 % sequence identity to SEQ ID NO : 20 , and a light chain ( LC ) having at least 95 % sequence identity to SEQ ID NO : .

24. An antibody which binds an epitope of XCL1 comprising one or more amino acids of SEQ ID NO : 27 .

25. The antibody of claim 24 , which binds two or more amino acids of SEQ ID NO : 27 . - 36 -

26. The antibody of claim 25 , which binds three or more amino acids of SEQ ID NO : 27 .

27. The antibody of claim 26 , which binds one or more amino acids of SEQ ID NO : 29 .

28. An antibody which binds an epitope of XCL1 comprising one or more amino acids of SEQ ID NO : 28 .

29. The antibody of claim 28 , which binds two or more amino acids of SEQ ID NO : 28 .

30. An antibody which binds an epitope of XCL1 comprising one or more amino acids of SEQ ID NO : 27 and one or more amino acids of SEQ ID NO : 28 .

31. The antibody of claim 30 , which binds two or more amino acids of SEQ ID NO : 27 and two or more amino acids of SEQ ID NO : 28 .

32. The antibody of claim 31 , which binds one or more amino acids of SEQ ID NO : 29 .

33. The antibody of any one of claims 1-32 , wherein the antibody has a human IgG1 or IgGisotype .

34. A pharmaceutical composition comprising an antibody of any one of claims 1-33 and one or more pharmaceutically acceptable carriers , diluents or excipients .

35. The antibody of any one of claims 1-33 , or the pharmaceutical composition of claim 34 , for use in therapy .

36. The antibody of any one of claims 1-33 , or the pharmaceutical composition of claim 34 , for use in the treatment of alopecia areata , vitiligo , asthma , psoriasis , multiple sclerosis , rheumatoid arthritis , or ulcerative colitis . - 37 -

37. A nucleic acid comprising a sequence encoding SEQ ID NO : 23 , SEQ ID NO : 24 , SEQ ID NO : 25 , or SEQ ID NO : 26 .

38. A vector comprising a first nucleic acid sequence encoding SEQ ID NO : 23 or SEQ ID NO : and a second nucleic acid sequence encoding SEQ ID NO : 24 or SEQ ID NO : 26 .

39. A cell comprising the vector of claim 38 .

40. A method of producing an antibody comprising culturing the cell of claim 39 under conditions such that the antibody is expressed and recovering the expressed antibody .

41. An antibody produced by culturing the cell of claim 39 under conditions such that the antibody is expressed and recovering the expressed antibody .