Methods of expanding and assessing b cells and using expanded b cells to treat disease

Abstract

Provided herein are methods of expanding B cells, and in particularly B10 cells capable of producing IL-10, ex vivo. The methods include incubation of harvested B cells in the presence of IL-21. Compositions comprising the ex vivo expanded B cells and methods of using the expanded B cell-containing compositions to treat diseases or conditions are also provided. Methods of assessing B10 cell function in a subject are also provided.

1 .- 21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . A method of treating a subject having an autoimmune disorder comprising administering a therapeutically effective amount of a composition comprising B10 cells cultured in vitro with IL-21, wherein the B10 cells are more than 85% of the total B cells in the composition and wherein said B10 cells are capable of producing IL-10 to a subject in need of treatment for an autoimmune disorder. 
     
     
         25 . The method of  claim 24 , wherein the autoimmune disease is selected from multiple sclerosis, lupus, arthritis, inflammatory bowel disease and scleroderma. 
     
     
         26 . The method of  claim 24 , wherein the autoimmune disease is selected from allergic contact dermatitis, allergic reactions to drugs, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune diseases of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, Behcet's disease, bullous pemphigoid and associated skin diseases, cardiomyopathy, Celiac disease, Celiac sprue-dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatrical pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, cutaneous necrotizing venulitis, discoid lupus, erythema multiforme, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic/autoimmune thrombocytopenia purpura (ITP), immunologic lung disease, immunologic renal disease, IgA neuropathy, juvenile arthritis, lichen planus, Meniere's disease, mixed connective tissue disease, type 1 or immune-mediated diabetes mellitus, myasthenia gravis, pemphigus-related disorders (e.g., pemphigus vulgaris), pernicious anemia, polyarteritis  nodosa , polychrondritis, polyglandular syndromes, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynauld's phenomenon, Reiter's syndrome, Rheumatoid arthritis, rheumatic fever, sarcoidosis, Sjögren's syndrome, stiff-man syndrome, spondyloarthropathies, systemic lupus erythematosis (SLE), lupus erythematosus, systemic vasculitis, takayasu arteritis, temporal arteristis/giant cell arteritis, thrombocytopenia, thyroiditis, ulcerative colitis, uveitis, vasculitides such as dermatitis herpetiformis vasculitis, vitiligo, and Wegener's granulomatosis. 
     
     
         27 . A method of treating a subject to prevent or treat organ, tissue or cell transplant rejection or associated chronic graft versus host disease or treating a subject receiving recombinant, therapeutic or xenogeneic protein(s) comprising administering a therapeutically effective amount of a composition comprising B10 cells cultured in vitro with IL-21, wherein the B10 cells are more than 85% of the total B cells in the composition and wherein said B10 cells are capable of producing IL-10 to a subject in need of treatment for transplant rejection, graft versus host disease or other disorder associated with receipt of a transplant or protein treatment. 
     
     
         28 . A method of treating a subject having an allergic disorder or inflammatory disorder comprising administering a therapeutically effective amount of a composition comprising B10 cells cultured in vitro with IL-21, wherein the B10 cells are more than 85% of the total B cells in the composition and wherein said B10 cells are capable of producing IL 10 to a subject in need of treatment for allergies or for inflammation. 
     
     
         29 . (canceled) 
     
     
         30 . The method of  claim 28 , wherein the inflammatory disorder is selected from asthma, encephilitis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), allergic disorders, septic shock, pulmonary fibrosis, undifferentitated spondyloarthropathy, undifferentiated arthropathy, arthritis, inflammatory osteolysis, and chronic inflammation resulting from chronic viral or bacterial infections. 
     
     
         31 . (canceled) 
     
     
         32 . The method of  claim 24 , wherein the composition comprises autologous B10 cells. 
     
     
         33 . (canceled) 
     
     
         34 . The method of  claim 24 , wherein the composition is administered after the onset of symptoms in the subject. 
     
     
         35 .- 49 . (canceled) 
     
     
         50 . The method of  claim 24 , wherein the composition comprises between 10 6  and 10 10  B10 cells. 
     
     
         51 . The method of  claim 24 , wherein the B10 cells were cultured in vitro with a CD40 agonist. 
     
     
         52 . The method of  claim 51 , wherein the CD40 agonist is CD154, a fragment of CD154, or antibody, aptamer or polypeptide, or fragment thereof reactive with CD40. 
     
     
         53 . The method of  claim 24 , wherein the B10 cells were cultured in vitro with a B cells survival promoter. 
     
     
         54 . The method of  claim 53 , wherein the B cell survival promoter is selected from at least one of feeder cells, BAFF (BLyS), BAFF fragments, APRIL, CD22 ligand, CD22 monoclonal antibody, or fragments thereof. 
     
     
         55 . The method of  claim 24 , wherein the B10 cells were cultured on feeder cells expressing a CD40 agonist and a B cell survival promoter. 
     
     
         56 . The method of  claim 55 , wherein the feeder cells are fibroblast, endothelial cells, epithelial cells, keratinocytes, melanocytes, or other mesenchymal or stromal cells. 
     
     
         57 . The method of  claim 24 , wherein the B10 cells were cultured in vitro with IL-4. 
     
     
         58 . The method of  claim 28 , wherein the composition comprises autologous B10 cells. 
     
     
         59 . The method of  claim 28 , wherein the B10 cells were cultured in vitro with a CD40 agonist. 
     
     
         60 . The method of  claim 28 , wherein the B10 cells were cultured in vitro with a B cells survival promoter. 
     
     
         61 . The method of  claim 28 , wherein the B10 cells were cultured in vitro with IL-4