HUMANIZED ANTI-CD40 ANTIBODIES AND USES THEREOF

1 . A humanized anti-CD40 antibody, or an antigen-binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises three complementarity determining regions (CDRs), CDR1, CDR2 and CDR3, having amino acid sequences about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively, and wherein the light chain variable region comprises three CDRs, CDR1, CDR2 and CDR3, having amino acid sequences about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NOs: 16, 17 and 18, respectively.

2 . A humanized anti-CD40 antibody, or an antigen-binding portion thereof, comprising a heavy chain variable region, wherein the heavy chain variable region comprises three CDRs, CDR1, CDR2 and CDR3, having amino acid sequences about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NOs: 13, 14 and 15, respectively.

3 . A humanized anti-CD40 antibody, or an antigen-binding portion thereof, comprising a light chain variable region, wherein the light chain variable region comprises three CDRs, CDR1, CDR2 and CDR3, having amino acid sequences about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NOs: 16, 17 and 18, respectively.

4 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the dissociation constant (K D ) of the antibody, or antigen-binding portion thereof, is less than about 1×10 −9 M.

5 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 11, 19, 20, 21, 24, 25 and 26, and wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 12, 22, 23, 27, 28 and 29.

6 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 21, and wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 23.

7 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 11, 19, 20, 21, 24, 25 and 26.

8 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 12, 22, 23, 27, 28 and 29.

9 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 19, 20 and 21, and wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to either of the amino acid sequences set forth in SEQ ID NOs: 22 and 23.

10 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 24, 25 and 26, and wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 27, 28 and 29.

11 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the antibody or antigen-binding portion thereof is selected from the group consisting of: (a) a whole immunoglobulin molecule; (b) an scFv; (c) a Fab fragment; (d) an F(ab′)2; and (e) a disulfide linked Fv.

12 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the antibody or antigen-binding portion thereof comprises at least one constant domain selected from the group consisting of: a) an IgG constant domain; and (b) an IgA constant domain.

13 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the antibody or antigen-binding portion thereof comprises at least one human constant domain.

14 . The antibody or antigen-binding portion thereof of any of claims 1-3 , wherein the antibody or antigen-binding portion thereof binds to CD40 extracellular domain.

15 . A humanized anti-CD40 antibody or an antigen-binding portion thereof, comprising a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to the amino acid sequence set forth in SEQ ID NOs: 11, 19, 20, 21, 24, 25 or 26.

16 . A humanized anti-CD40 antibody, or an antigen-binding portion thereof, comprising a light chain variable region, wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to the amino acid sequence set forth in SEQ ID NOs: 12, 22, 23, 27, 28 or 29.

17 . A humanized anti-CD40 antibody, or an antigen-binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 11, 19, 20, 21, 24, 25 and 26, and wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to any one of the amino acid sequences set forth in SEQ ID NOs: 12, 22, 23, 27, 28 and 29.

18 . A humanized or chimeric anti-CD40 antibody, or an antigen-binding portion thereof, comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 21, and wherein the light chain variable region comprises an amino acid sequence about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 23.

19 . The antibody or antigen-binding portion thereof of any of claims 1-18 , wherein the CD40 is human or rhesus CD40.

20 . The antibody or antigen-binding portion thereof of any of claims 1-18 , wherein the antibody or antigen-binding portion thereof blocks B lymphocyte activation by CD154-expressing Jurkat cells in vitro.

21 . The antibody or antigen-binding portion thereof of any of claims 1-18 , wherein the antibody or antigen-binding portion thereof inhibits B lymphocyte CD23, CD80, or CD86 expression.

22 . A composition comprising the antibody or antigen-binding portion thereof of any of claims 1-21 , and at least one pharmaceutically acceptable carrier.

23 . A polynucleotide encoding the antibody or antigen-binding portion thereof of any of claims 1-21 .

24 . A vector comprising the polynucleotide of claim 23 .

25 . A cell comprising the vector of claim 24 .

26 . A method of suppressing the immune system in a subject, comprising the step of administering to the subject an effective amount of the antibody or antigen-binding portion thereof of any of claims 1-21 .

27 . A method of treating or treating prophylactically transplant rejection, or increasing the duration of time before transplant rejection occurs, in a subject in need thereof, the method comprising the step of administering to the subject an effective amount of the antibody or antigen-binding portion thereof of any of claims 1-21 .

28 . The method of claim 26 or 27 , wherein the subject has received, or is in need of, an organ transplantation.

29 . The method of claim 28 , wherein the organ is selected from the group consisting of heart, kidney, lung, liver, pancreas, intestine, and thymus, or a portion thereof.

30 . The method of claim 26 or 27 , wherein the subject has received, or is in need of, a tissue transplantation.

31 . The method of claim 30 , wherein the tissue is bone, tendon, cornea, skin, heart valve, vein, or bone marrow.

32 . The method of any of claims 26-31 , wherein the administration is commenced prior to the transplantation.

33 . The method of any of claims 26-31 , wherein the administration continues for at least one month following the transplantation.

34 . The method of claim 33 , wherein the administration continues for at least six months following said transplantation of said graft.

35 . A method of treating or treating prophylactically graft-versus-host disease in a subject in need thereof, the method comprising the step of administering to the subject an effective amount of the antibody or antigen-binding portion thereof of any of claims 1-21 .

36 . A method of treating or treating prophylactically an autoimmune disorder in a subject in need thereof, the method comprising the step of administering to the subject an effective amount of the antibody or antigen-binding portion thereof of any of claims 1-21 .

37 . The method of claim 36 , wherein the autoimmune disorder is associated with or caused by the presence of an autoantibody.

38 . The method of claim 36 , wherein the autoimmune disorder is selected from the group consisting of systemic lupus erythematosus (SLE), CREST syndrome (calcinosis, Raynaud's syndrome, esophageal dysmotility, sclerodactyl, and telangiectasia), opsoclonus, inflammatory myopathy (e.g., polymyositis, dermatomyositis, and inclusion-body myositis), systemic scleroderma, primary biliary cirrhosis, celiac disease (e.g., gluten sensitive enteropathy), dermatitis herpetiformis, Miller-Fisher Syndrome, acute motor axonal neuropathy (AMAN), multifocal motor neuropathy with conduction block, autoimmune hepatitis, antiphospholipid syndrome, Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, rheumatoid arthritis, chronic autoimmune hepatitis, scleromyositis, myasthenia gravis, Lambert-Eaton myasthenic syndrome, Hashimoto's thyroiditis, Graves' disease, Paraneoplastic cerebellar degeneration, Stiff person syndrome, limbic encephalitis, Isaacs Syndrome, Sydenham's chorea, pediatric autoimmune neuropsychiatric disease associated with Streptococcus (PANDAS), encephalitis, diabetes mellitus type 1, and Neuromyelitis optica.

39 . The method of claim 36 , wherein the autoimmune disorder is selected from the group consisting of pernicious anemia, Addison's disease, psoriasis, inflammatory bowel disease, psoriatic arthritis, Sjögren's syndrome, lupus erythematosus (e.g., discoid lupus erythematosus, drug-induced lupus erythematosus, and neonatal lupus erythematosus), multiple sclerosis, and reactive arthritis.

40 . The method of claim 36 , wherein the autoimmune disorder is selected from the group consisting of polymyositis, dermatomyositis, multiple endocrine failure, Schmidt's syndrome, autoimmune uveitis, adrenalitis, thyroiditis, autoimmune thyroid disease, gastric atrophy, chronic hepatitis, lupoid hepatitis, atherosclerosis, presenile dementia, demyelinating diseases, subacute cutaneous lupus erythematosus, hypoparathyroidism, Dressler's syndrome, autoimmune thrombocytopenia, idiopathic thrombocytopenic purpura, hemolytic anemia, pemphigus vulgaris, pemphigus, alopecia arcata, pemphigoid, scleroderma, progressive systemic sclerosis, adult onset diabetes mellitus (e.g., type II diabetes), male and female autoimmune infertility, ankylosing spondolytis, ulcerative colitis, Crohn's disease, mixed connective tissue disease, polyarteritis nedosa, systemic necrotizing vasculitis, juvenile onset rheumatoid arthritis, glomerulonephritis, atopic dermatitis, atopic rhinitis, Goodpasture's syndrome, Chagas' disease, sarcoidosis, rheumatic fever, asthma, recurrent abortion, anti-phospholipid syndrome, farmer's lung, erythema multiforme, post cardiotomy syndrome, Cushing's syndrome, autoimmune chronic active hepatitis, bird-fancier's lung, allergic disease, allergic encephalomyelitis, toxic epidermal necrolysis, alopecia, Alport's syndrome, alveolitis, allergic alveolitis, fibrosing alveolitis, interstitial lung disease, erythema nodosum, pyoderma gangrenosum, transfusion reaction, leprosy, malaria, leishmaniasis, trypanosomiasis, Takayasu's arteritis, polymyalgia rheumatica, temporal arteritis, schistosomiasis, giant cell arteritis, ascariasis, aspergillosis, Sampter's syndrome, eczema, lymphomatoid granulomatosis, Behcet's disease, Caplan's syndrome, Kawasaki's disease, dengue, endocarditis, endomyocardial fibrosis, endophthalmitis, erythema elevatum et diutinum, erythroblastosis fetalis, eosinophilic faciitis, Shulman's syndrome, Felty's syndrome, filariasis, cyclitis, chronic cyclitis, heterochronic cyclitis, Fuch's cyclitis, IgA nephropathy, Henoch-Schonlein purpura, graft versus host disease, transplantation rejection, human immunodeficiency virus infection, echovirus infection, cardiomyopathy, Alzheimer's disease, parvovirus infection, rubella virus infection, post vaccination syndromes, congenital rubella infection, Hodgkin's and non-Hodgkin's lymphoma, renal cell carcinoma, multiple myeloma, Eaton-Lambert syndrome, relapsing polychondritis, malignant melanoma, cryoglobulinemia, Waldenstrom's macroglobulemia, Epstein-Barr virus infection, mumps, Evan's syndrome, and autoimmune gonadal failure.

41 . The method of any of claims 26-40 , wherein the subject is a human.

42 . The method of any of claims 26-40 , wherein the administration is parenteral, intravenous, subcutaneous, intramuscular, transdermal, oral, topical, intrathecal, or local.

43 . The method of any of claims 26-40 , wherein the method further comprises administration of an immunosuppressant within six months of the administration of the antibody or antigen-binding portion thereof.

44 . The method of claim 43 , wherein the immunosuppressant is selected from the group consisting of a calcineurin inhibitor, tacrolimus, an mTor inhibitor, fingolimod, myriocin, alemtuzumab, rituximab, an anti-CD4 monoclonal antibody, an anti-LFA1 monoclonal antibody, an anti-LFA3 monoclonal antibody, an anti-CD45 antibody, an anti-CD19 antibody, monabatacept, belatacept, indolyl-ASC; azathioprine, lymphocyte immune globulin and anti-thymocyte globulin [equine], mycophenolate mofetil, mycophenolate sodium, daclizumab, basiliximab, cyclophosphamide, prednisone, prednisolone, leflunomide, FK778, FK779, 15-deoxyspergualin, busulfan, fludarabine, methotrexate, 6-mercaptopurine, 15-deoxyspergualin, LF15-0195, bredinin, brequinar, and muromonab-CD3.

45 . The method of claim 44 , wherein the calcineurin inhibitor is cyclosporin A or cyclosporine G.

46 . The method of claim 44 , wherein the mTor inhibitor is sirolimus, temsirolimus, zotarolimus, or everolimus.

47 . The method of claim 44 , wherein the anti-CD45 antibody is an anti-CD45RB antibody.

48 . The method of claim 44 , wherein the immunosuppressant is belatacept.

49 . The method of claim 44 , wherein the antibody or antigen-binding portion thereof and the immunosuppressant are administered within one month of each other.

50 . The method of claim 49 , wherein the antibody or antigen-binding portion thereof and the immunosuppressant are administered within one week of each other.

51 . An isolated polypeptide comprising the antibody or antigen-binding portion thereof of any of claims 1-21 .

52 . A method of producing the antibody or antigen-binding portion thereof of any of claims 1-21 , the method comprising the steps of:

(a) culturing the cells of claim 25 in culture medium under conditions wherein the polynucleotide is expressed, thereby producing at least one polypeptide comprising the antibody or antigen-binding portion thereof; and

(b) recovering the polypeptide from the cells or culture medium.