Antibodies to MAdCAM

Abstract

The present invention relates to antibodies including human antibodies and antigen-binding portions thereof that specifically bind to MAdCAM, preferably human MAdCAM and that function to inhibit MAdCAM. The invention also relates to human anti-MAdCAM antibodies and antigen-binding portions thereof. The invention also relates to antibodies that are chimeric, bispecific, derivatized, single chain antibodies or portions of fusion proteins. The invention also relates to isolated heavy and light chain immunoglobulins derived from human anti-MAdCAM antibodies and nucleic acid molecules encoding such immunoglobulins. The present invention also relates to methods of making human anti-MAdCAM antibodies, compositions comprising these antibodies and methods of using the antibodies and compositions for diagnosis and treatment. The invention also provides gene therapy methods using nucleic acid molecules encoding the heavy and/or light immunoglobulin molecules that comprise the human anti-MAdCAM antibodies. The invention also relates to transgenic animals or plants comprising nucleic acid molecules of the invention.

1 . (canceled) 
     
     
         2 . The human monoclonal antibody or antigen-binding portion according to  claim 3 , wherein said antibody or portion possesses at least one of the following properties:
 (a) binds to human cells;   (b) has a selectivity for MAdCAM over VCAM or fibronectin of at least 100 fold; or   (c) inhibits the recruitment of lymphocytes to gastrointestinal lymphoid tissue.   
     
     
         3 . A human monoclonal antibody or an antigen-binding portion thereof, wherein said antibody or portion binds human Mucosal Adressin Cell Adhesion Molecule (MAdCAM) with a K d  of 3×10 −10  M or less and inhibits α 4 β 7  binding to human MAdCAM. 
     
     
         4 . A hybridoma cell line that produces the human monoclonal antibody according to  claim 1 , wherein the hybridoma is selected from the group consisting of 1.7.2 (ECACC Accession No. 03090901), 1.8.2 (ECACC Accession No. 03090902), 6.1-4.2 (ECACC Accession No. 03090903), 6.2-2.2 (ECACC Accession No. 03090904), 6.3-4.2 (ECACC Accession No. 03090905), 6.6-7.1 (ECACC Accession No. 03090906), 6.7-3.2 (ECACC Accession No. 03090907), 6.7-7.1 (ECACC Accession No. 03090908), 7.1-6.6 (ECACC-Accession No. 03090909), 7.2-0.5 (ECACC Accession No. 03090910), 7.2-6.4 (ECACC Accession No. 03090911), and 9.8.2 (ECACC Accession No. 03090912). 
     
     
         5 . The human monoclonal antibody produced by the hybridoma cell line according to  claim 4  or an antigen-binding portion of said monoclonal antibody. 
     
     
         6 . The human monoclonal antibody according to  claim 5 , wherein the heavy chain C-terminal lysine is cleaved. 
     
     
         7 . The human monoclonal antibody or antigen-binding portion according to  claim 3  wherein the antibody or portion thereof has at least one of the following properties:
 (a) cross-competes with a reference antibody for binding to MAdCAM;   (b) competes with a reference antibody for binding to MAdCAM;   (c) binds to the same epitope of MAdCAM as a reference antibody;   (d) binds to MAdCAM with substantially the same K d  as a reference antibody; or   (e) binds to MAdCAM with substantially the same off rate as a reference antibody;   wherein the reference antibody is selected from the group consisting of: monoclonal antibody 1.7.2, monoclonal antibody 1.8.2, monoclonal antibody 6.14.2, monoclonal antibody 6.22.2, monoclonal antibody 6.34.2, monoclonal antibody 6.67.1, monoclonal antibody 6.73.2, monoclonal antibody 6.77.1, monoclonal antibody 7.16.6, monoclonal antibody 7.20.5, monoclonal antibody 7.26.4, monoclonal antibody 9.8.2, monoclonal antibody 6.22.2-mod, monoclonal antibody 6.34.2-mod, monoclonal antibody 6.67.1-mod, monoclonal antibody 6.77.1-mod and monoclonal antibody 7.26.4-mod.   
     
     
         8 . A monoclonal antibody or an antigen-binding portion thereof that specifically binds MAdCAM, wherein the antibody is selected from the group consisting of:
 (a) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 2 and SEQ ID NO: 4, without the signal sequences;   (b) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 6 and SEQ ID NO: 8, without the signal sequences;   (c) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 12, without the signal sequences;   (d) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 14 and SEQ ID NO: 16, without the signal sequences;   (e) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 18 and SEQ ID NO: 20, without the signal sequences;   (f) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 22 and SEQ ID NO: 24, without the signal sequences;   (g) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 26 and SEQ ID NO: 28, without the signal sequences;   (h) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 30 and SEQ ID NO: 32, without the signal sequences;   (i) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 34 and SEQ ID NO: 36, without the signal sequences;   (j) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 38 and SEQ ID NO: 40, without the signal sequences;   (k) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 42 and SEQ ID NO: 44, without the signal sequences;   (l) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 46 and SEQ ID NO: 48, without the signal sequences;   (m) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 52 and SEQ ID NO: 54, without the signal sequences;   (n) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 56 and SEQ ID NO: 58, without the signal sequences;   (o) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 60 and SEQ ID NO: 62, without the signal sequences;   (p) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 64 and SEQ ID NO: 66, without the signal sequences; and   (q) an antibody comprising the amino acid sequences set forth in SEQ ID NO: 42 and SEQ ID NO: 68, without the signal sequences.   
     
     
         9 . A monoclonal antibody or an antigen-binding portion thereof, wherein the heavy chain of said antibody comprises the heavy chain CDR1, CDR2 and CDR3 or wherein the light chain comprises the light chain CDR1, CDR2 and CDR3 of a monoclonal antibody selected from the group consisting of: 1.7.2, 1.8.2, 6.14.2, 6.22.2, 6.34.2, 6.67.1, 6.73.2, 6.77.1, 7.16.6, 7.20.5, 7.26.4, 9.8.2, 6.22.2-mod, 6.34.2-mod, 6.67.1-mod, 6.77.1-mod and 7.26.4-mod. 
     
     
         10 . The monoclonal antibody or antigen-binding portion according to  claim 9 , wherein said antibody or portion comprises a heavy chain that utilizes a human VH 1-18 gene, a human VH 3-15 gene, a human VH 3-21 gene, a human VH 3-23 gene, a human VH 3-30 gene, a human VH 3-33 gene or a human VH 4-4 gene. 
     
     
         11 . The monoclonal antibody or an antigen-binding portion thereof according to  claim 10 , wherein said antibody or portion comprises a light chain that utilizes a human VK A2 gene, a human VK A3 gene, a human VK A26 gene, a human VK B3 gene, a human VK 012 gene or a human VK 018 gene. 
     
     
         12 . The human monoclonal antibody or antigen-binding portion according to  claim 1 , wherein the heavy chain variable region, the light chain variable region or both are at least 90% identical in amino acid sequence to the corresponding region or regions of a monoclonal antibody selected from the group consisting of: monoclonal antibody 1.7.2, monoclonal antibody 1.8.2, monoclonal antibody 6.14.2, monoclonal antibody 6.22.2, monoclonal antibody 6.34.2, monoclonal antibody 6.67.1, monoclonal antibody 6.73.2, monoclonal antibody 6.77.1, monoclonal antibody 7.16.6, monoclonal antibody 7.20.5, monoclonal antibody 7.26.4 monoclonal antibody 9.8.2, monoclonal antibody 6.22.2-mod, monoclonal antibody 6.34.2-mod, monoclonal antibody 6.67.1-mod, monoclonal antibody 6.77.1-mod and monoclonal antibody 7.26.4-mod. 
     
     
         13 . A monoclonal antibody or an antigen-binding portion thereof that specifically binds MAdCAM, wherein:
 (a) the heavy chain comprises the heavy chain CDR1, CDR2 and CDR3 amino acid sequences of a reference antibody selected from the group consisting of: 1.7.2, 1.8.2, 6.14.2, 6.22.2, 6.34.2, 6.67.1, 6.73.2, 6.77.1, 7.16.6, 7.20.5, 7.26.4, 9.8.2, 6.22.2-mod, 6.34.2-mod, 6.67.1-mod, 6.77.1-mod and 7.26.4-mod;   (b) the light chain comprises the light chain CDR1, CDR2 and CDR3 amino acid sequences of a reference antibody selected from the group consisting of: 1.7.2, 1.8.2, 6.14.2, 6.22.2, 6.34.2, 6.67.1, 6.73.2, 6.77.1, 7.16.6, 7.20.5, 7.26.4, 9.8.2, 6.22.2-mod, 6.34.2-mod, 6.67.1-mod, 6.77.1-mod and 7.26.4-mod;   (c) the antibody comprises a heavy chain of (a) and a light chain of (b); and   (d) the antibody of (c) wherein the heavy chain and light chain CDR amino acid sequences are selected from the same reference antibody   
     
     
         14 . The monoclonal antibody or antigen-binding portion according to  claim 13 , wherein the heavy chain, the light chain or both comprise the amino acid sequence from the beginning of the CDR1 through the end of the CDR3 of the heavy chain, the light chain or both, respectively, of the reference antibody. 
     
     
         15 . The monoclonal antibody or antigen-binding portion according to  claim 13 , wherein said antibody comprises:
 (a) a heavy chain comprising the heavy chain variable region amino acid sequence of an antibody selected from the group consisting of: 1.7.2 (SEQ ID NO: 2); 1.8.2 (SEQ ID NO: 6); 6.14.2 (SEQ ID NO: 10); 6.22.2 (SEQ ID NO: 14); 6.34.2 (SEQ ID NO: 18); 6.67.1 (SEQ ID NO: 22); 6.73.2 (SEQ ID NO: 26); 6.77.1 (SEQ ID NO: 30); 7.16.6 (SEQ ID NO: 34); 7.20.5 (SEQ ID NO: 38); 7.26.4 (SEQ ID NO: 42); and 9.8.2 (SEQ ID NO: 46); 6.22.2-mod (SEQ ID NO: 52); 6.34.2-mod (SEQ ID NO: 56); 6.67.1-mod (SEQ ID NO: 60); 6.77.1-mod (SEQ ID NO: 64); and 7.26.4-mod (SEQ ID NO: 42);   (b) a light chain comprising the light chain variable region amino acid sequence of an antibody selected from the group consisting of: 1.7.2 (SEQ ID NO: 4); 1.8.2 (SEQ ID NO: 8); 6.14.2 (SEQ ID NO: 12); 6.22.2 (SEQ ID NO: 16); 6.34.2 (SEQ ID NO: 20); 6.67.1 (SEQ ID NO: 24); 6.73.2 (SEQ ID NO: 28); 6.77.1 (SEQ ID NO: 32); 7.16.6 (SEQ ID NO: 36); 7.20.5 (SEQ ID NO: 40); 7.26.4 (SEQ ID NO: 44); and 9.8.2 (SEQ ID NO: 48); 6.22.2-mod (SEQ ID NO: 54); 6.34.2-mod (SEQ ID NO: 58); 6.67.1-mod (SEQ ID NO: 62); 6.77.1-mod (SEQ ID NO: 66); and 7.26.4-mod (SEQ ID NO: 68); or   (c) the heavy chain of (a) and the light chain of (b).   
     
     
         16 . The monoclonal antibody according to  claim 3  that is an immunoglobulin G (IgG), an IgM, an IgE, an IgA or an IgD molecule, a humanized antibody, a chimeric antibody or a bispecific antibody. 
     
     
         17 . The antigen-binding portion according to  claim 3  which is an Fab fragment, an F(ab′) 2  fragment, an F V  fragment or a single chain antibody. 
     
     
         18 . A pharmaceutical composition comprising an effective amount of the monoclonal antibody or antigen-binding portion according to  claim 3  and a pharmaceutically acceptable carrier. 
     
     
         19 . A method of treating inflammatory disease in a subject in need thereof, comprising the step of administering to said subject the monoclonal antibody or antigen-binding portion according to  claim 3  wherein said antibody or antigen-binding portion inhibits binding of MAdCAM to α 4 β 7 . 
     
     
         20 . The method of  claim 19 , wherein the inflammatory disease is inflammatory disease of the gastrointestinal tract. 
     
     
         21 . The method of  claim 20 , wherein the inflammatory disease of the gastrointestinal tract is selected from the group consisting of inflammatory bowel disease, Crohn's disease, ulcerative colitis, diverticula disease, gastritis, liver disease, primary biliary sclerosis and sclerosing cholangitis. 
     
     
         22 . (canceled) 
     
     
         23 . The method of  claim 20 , wherein the inflammatory disease is insulin-dependent diabetes or graft versus host disease. 
     
     
         24 . An isolated cell line that produces the monoclonal antibody or antigen-binding portion according to  claim 3  or the heavy chain or light chain of said antibody or of said portion thereof. 
     
     
         25 . The cell line according to  claim 24  that produces an antibody selected from the group consisting of: 1.7.2, 1.8.2, 6.14.2, 6.22.2, 6.34.2, 6.67.1, 6.73.2, 6.77.1, 7.16.6, 7.20.5, 7.26.4 and 9.8.2, or an antibody comprising the amino acid sequences of one of said antibodies. 
     
     
         26 . The cell line according to  claim 25  that produces a monoclonal antibody selected from the group consisting of: 6.22.2-mod, 6.34.2-mod, 6.67.1-mod, 6.77.1-mod and 7.26.4-mod or an antibody comprising the amino acid sequences of one of said antibodies. 
     
     
         27 . An isolated nucleic acid molecule comprising a nucleotide sequence that encodes the heavy chain or an antigen-binding portion thereof or the light chain or an antigen-binding portion thereof of an antibody according to  claim 3 . 
     
     
         28 . A vector comprising the nucleic acid molecule according to  claim 27 , wherein the vector optionally comprises an expression control sequence operably linked to the nucleic acid molecule. 
     
     
         29 . A host cell comprising the vector according to  claim 28 . 
     
     
         30 . A host cell comprising a nucleic acid molecule encoding the heavy chain or an antigen-binding portion thereof and a nucleic acid molecule encoding the light chain or an antigen-binding portion thereof of an antibody or antigen-binding portion according to  claim 3 . 
     
     
         31 . A method for producing a human monoclonal antibody or antigen-binding portion thereof that specifically binds MAdCAM, comprising culturing the host cell according to  claim 29  under suitable conditions and recovering said antibody or antigen-binding portion. 
     
     
         32 . A non-human transgenic animal or transgenic plant comprising (a) nucleic acid molecule encoding the heavy chain or an antigen-binding portion thereof; (b) a nucleic acid molecule encoding the light chain or an antigen-binding portion thereof; or (c) both (a) and (b) of an antibody according to  claim 3 , wherein the non-human transgenic animal or transgenic plant expresses said heavy chain or light chain or both. 
     
     
         33 . A method of isolating an antibody or antigen-binding portion thereof that specifically binds to MAdCAM, comprising the step of isolating the antibody from the non-human transgenic animal or transgenic plant according to  claim 32 . 
     
     
         34 . A method of treating a subject in need thereof with the monoclonal antibody to  claim 3 , comprising the steps of:
 (a) administering an effective amount of an isolated nucleic acid molecule encoding the heavy chain or an antigen-binding portion of said antibody, an isolated nucleic acid molecule encoding the light chain or an antigen-binding portion of said antibody, or nucleic acid molecules encoding the light chain and the heavy chain or antigen-binding portions of said antibody; and   (b) expressing the nucleic acid molecule.   
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . A method of inhibiting α 4 β 7  binding to cells expressing human MAdCAM, comprising contacting the cells with the monoclonal antibody or portion according to  claim 3 . 
     
     
         38 . A method for inhibiting MAdCAM-mediated leukocyte-endothelial cell adhesion, comprising contacting the endothelial cells with the monoclonal antibody or antigen-binding portion according to  claim 3 . 
     
     
         39 . A method for inhibiting MAdCAM-mediated leukocyte adhesion, migration and infiltration into tissues, comprising the step of contacting the endothelial cells with the monoclonal antibody or antigen-binding portion according to  claim 3 . 
     
     
         40 . A method for inhibiting α 4 β 7 /MAdCAM-dependent cellular adhesion, comprising the step of contacting cells expressing human MAdCAM with the monoclonal antibody or antigen-binding portion according to  claim 3 . 
     
     
         41 . A method for inhibiting the MAdCAM-mediated recruitment of lymphocytes to gastrointestinal lymphoid tissue, comprising the step of contacting cells expressing human MAdCAM with the monoclonal antibody or antigen-binding portion according to  claim 3 . 
     
     
         42 . A monoclonal antibody or an antigen-binding portion thereof that specifically binds MAdCAM, wherein said antibody or portion thereof comprises one or more of an FR1, FR2, FR3 or FR4 amino acid sequence of a human monoclonal antibody selected from the group consisting of: 1.7.2, 1.8.2, 6.14.2, 6.22.2, 6.34.2, 6.67.1, 6.73.2, 6.77.1, 7.16.6, 7.20.5, 7.26.4 9.8.2, 6.22.2-mod, 6.34.2-mod, 6.67.1-mod, 6.77.1-mod and 7.26.4-mod. 
     
     
         43 . The human monoclonal antibody or antigen-binding portion according to  claim 3 , wherein the antibody comprises:
 (a) a heavy chain amino acid sequence that is at least 90% identical to the heavy chain amino acid sequence of a monoclonal antibody selected from the group consisting of: 1.7.2, 1.8.2, 6.14.2, 6.22.2, 6.34.2, 6.67.1, 6.73.2, 6.77.1, 7.16.6, 7.20.5, 7.26.4, 9.8.2, 6.22.2-mod, 6.34.2-mod, 6.67.1-mod, 6.77.1-mod and 7.26.4-mod;   (b) a light chain amino acid sequence that is at least 90% identical to the light chain amino acid sequence of a monoclonal antibody selected from the group consisting of: 1.7.2, 1.8.2, 6.14.2, 6.22.2, 6.34.2, 6.67.L, 6.73.2, 6.77.1, 7.16.6, 7.20.5, 7.26.4, 9.8.2, 6.22.2-mod, 6.34.2-mod, 6.67.1-mod, 6.77.1-mod and 7.26.4-mod;   (c) both (a) and (b); or   (d) either (a), (b) or (c), with or without the signal sequence.   
     
     
         44 . A method for diagnosing a disorder characterized by circulating soluble human MAdCAM comprising the steps of: (1) contacting a biological sample with the monoclonal antibody or antigen-binding portion according to  claim 3  and (2) detecting binding. 
     
     
         45 . A method for detecting inflammation in a subject comprising the steps of: (1) administering to said subject the monoclonal antibody or antigen-binding portion according to  claim 3  wherein said antibody or portion thereof is detectably labeled and (2) detecting binding. 
     
     
         46 . A diagnostic kit comprising the monoclonal antibody or antigen-binding portion according to  claim 3 . 
     
     
         47 . The pharmaceutical composition according to  claim 18  further comprising one or more additional anti-inflammatory or immunomodulatory agents. 
     
     
         48 . The pharmaceutical composition according to  claim 47 , wherein the one or more additional anti-inflammatory or immunomodulatory agents are selected from the group consisting of: corticosteroids, aminosalicylates, azathioprine, methotrexate, cyclosporin, FK506, IL-10, GM-CSF, rapamycin, anti-TNFα agents and adhesion molecule antagonists. 
     
     
         49 . (canceled) 
     
     
         50 . (canceled) 
     
     
         51 . A method of detecting the effect of administration of an inhibitory anti-MAdCAM antibody or antigen-binding portion thereof to a subject comprising the steps of:
 (a) administering to a subject a human monoclonal antibody that specifically binds to MAdCAM; and   (b) determining whether there is an increase in the levels of circulating α 4 β 7 -expressing leukocytes.   
     
     
         52 . The method according to  claim 51 , wherein said leukocytes are lymphocytes. 
     
     
         53 . The method according to  claim 51 , wherein said increase in the levels of circulating α 4 β 7 -expressing leukocytes is determined by FACS analysis.