Cytokines and neuroantigens for treatment of immune disorders

Abstract

The present invention provides methods of regulating an immunological disorder comprising administering to a subject an effective amount of (i) an autoimmune antigen in conjunction with (ii) an anti-inflammatory cytokine. Compositions including the same are also provided.

1 . A method of regulating an immunological disorder comprising administering to a subject an effective amount of (i) an autoimmune antigen or portion thereof in conjunction with (ii) an anti-inflammatory cytokine, with the proviso that (i) and (ii) do not comprise a fusion protein. 
     
     
         2 . The method of  claim 1 , wherein the autoimmune antigen is selected from the group consisting of myelin basic protein (MBP), proteolipid protein (PLP), myelin oligodendrocyte glycoprotein (MOG), myelin-associated oligodendrocytic basic protein and cardiac myosin, or portions thereof. 
     
     
         3 . The method of  claim 1 , wherein the anti-inflammatory cytokine is IFN-β, and the autoimmune antigen or portion thereof is more than one autoimmune antigen selected from the group consisting of myelin basic protein (MBP), proteolipid protein (PLP), myelin oligodendrocyte glycoprotein (MOG), myelin-associated oligodendrocytic basic protein and cardiac myosin, or portions thereof. 
     
     
         4 . The method of  claim 1 , wherein the anti-inflammatory cytokine is IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17 and IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-28A, IL-28B, IL-29, IL-31, IL-32, IL-33, macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), erythropoietin, TNF-α, TGF-β, lymphotoxin, leptin, IFN-α, IFN-β, IFN-γ or a chemokine. 
     
     
         5 . The method of  claim 1 , wherein the immunological disorder is an autoimmune disease, an allergic or hypersensitivity disease, a transplantation and/or tissue disorder or combinations thereof. 
     
     
         6 . The method of  claim 5 , wherein the immunological disorder is an autoimmune disease affecting the nervous system, endocrine system, hematopoietic system, gastrointestinal tract, renal system, cardiac system, vascular system, musculoskeletal system or combinations thereof. 
     
     
         7 . The method of  claim 1 , wherein the immunological disorder is multiple sclerosis. 
     
     
         8 . The method of  claim 5 , wherein the allergic disease is allergic rhinitis, asthma, atopic dermatitis, allergic gastroenteropathy, contact dermatitis, drug allergy or combinations thereof. 
     
     
         9 . The method of  claim 5 , wherein the affected tissue is kidney, liver, pancreas, heart, lung, bone, skin or combinations thereof. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 1 , wherein (i) and (ii) are administered simultaneously or sequentially. 
     
     
         12 . The method of  claim 1 , wherein at least one of (i) and (ii) is administered parenterally. 
     
     
         13 . A fusion protein comprising:
 (i) an encephalitogenic determinant portion of a myelin basic protein, wherein the encephalitogenic determinant portion of the myelin basic protein is (1) an amino acid sequence of SEQ ID NO:2, (2) an amino acid sequence having at least 80% identity or homology with the amino acid sequence of SEQ ID NO:2, (3) an amino acid sequence encoded by a nucleic acid sequence encoding an encephalitogenic determinant portion of the myelin basic protein, or (4) an amino acid sequence encoded by a nucleic acid sequence that hybridizes with the complement of the nucleic acid sequence of (3) under stringent conditions as represented by hybridization conditions of 0.5M NaHPO 4 , 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at 65° C. and wash conditions of 0.1×SSC/0.1% SDS at 68° C.;   (ii) an enterokinase linking moiety, wherein the enterokinase moiety is (1) an amino acid sequence of SEQ ID NO:1, (2) an amino acid sequence having at least 80% identity or homology with the amino acid sequence of SEQ ID NO:1, (3) an amino acid sequence encoded by a nucleic acid sequence encoding an enterokinase recognition site, or (4) an amino acid sequence encoded by a nucleic acid sequence that hybridizes with the complement of the nucleic acid sequence of (3) under stringent conditions as represented by hybridization conditions of 0.5M NaHPO 4 , 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at 65° C. and wash conditions of 0.1×SSC/0.1% SDS at 68° C.; and   (iii) IFN-β, IFN-γ, GM-CSF, G-CSF, IL-16, IL-4 or IL-2.   
     
     
         14 . The fusion protein of  claim 13 , wherein the myelin basic protein comprises a C-terminal modification. 
     
     
         15 . The fusion protein of  claim 14 , wherein the C-terminal modification comprises a 6-histidine tag. 
     
     
         16 . An isolated nucleic acid encoding the fusion protein of  claim 13 . 
     
     
         17 . A vector comprising the isolated nucleic acid of  claim 16 . 
     
     
         18 . A host cell comprising the vector of  claim 17 . 
     
     
         19 . A method of making a fusion protein comprising:
 (a) culturing a host cell of  claim 18 ; and   (b) collecting the encoded fusion protein from said culture.   
     
     
         20 . A composition comprising:
 (a) an autoimmune antigen or portion thereof;   (b) an anti-inflammatory cytokine; and   (c) a pharmaceutically acceptable carrier, excipient or diluent, with the proviso that (a) and (b) do not comprise a fusion protein.   
     
     
         21 . A composition comprising:
 (a) at least one fusion protein comprising (i) an autoimmune antigen or portion thereof; (ii) optionally an enterokinase linking moiety, wherein the enterokinase linking moiety is (1) an amino acid sequence of SEQ ID NO:1, (2) an amino acid sequence having at least 80% identity or homology with the amino acid sequence of SEQ ID NO:1, (3) an amino acid sequence encoded by a nucleic acid sequence encoding an enterokinase recognition site, or (4) an amino acid sequence encoded by a nucleic acid sequence that hybridizes with the complement of the nucleic acid sequence of (3) under stringent conditions as represented by hybridization conditions of 0.5M NaHPO 4 , 7% sodium dodecyl sulfate (SDS), 1 mM EDTA at 65° C. and wash conditions of 0.1×SSC/0.1% SDS at 68° C.; and (iii) an anti-inflammatory cytokine;   (b) an anti-inflammatory cytokine; and   (c) a pharmaceutically acceptable carrier, excipient or diluent.   
     
     
         22 . The composition of  claim 21 , wherein at least one of the anti-inflammatory cytokines is IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17 and IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-28A, IL-28B, IL-29, IL-31, IL-32, IL-33, macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), erythropoietin, TNF-α, TGF-β, lymphotoxin, leptin, IFN-α, IFN-β, IFN-γ or a chemokine. 
     
     
         23 . The composition of  claim 20 , wherein the autoimmune antigen or portion thereof is selected from the group consisting of myelin basic protein (MBP), proteolipid protein (PLP), myelin oligodendrocyte glycoprotein (MOG), myelin-associated oligodendrocytic basic protein and cardiac myosin, or portions thereof. 
     
     
         24 . The composition of  claim 20 , wherein the anti-inflammatory cytokine is IFN-β, and the autoimmune antigen or portion thereof is selected from the group consisting of myelin basic protein (MBP), proteolipid protein (PLP), myelin oligodendrocyte glycoprotein (MOG) and myelin-associated oligodendrocytic basic protein, or portions thereof. 
     
     
         25 . The composition of  claim 21 , wherein the anti-inflammatory cytokine of the fusion protein is IFN-β, IFN-γ, GM-CSF, G-CSF, IL-16, IL-4 or IL-2, and the autoimmune antigen or portion thereof is selected from the group consisting of myelin basic protein (MBP), proteolipid protein (PLP), myelin oligodendrocyte glycoprotein (MOG) and myelin-associated oligodendrocytic basic protein, or portions thereof. 
     
     
         26 . A method of regulating an immunological disorder comprising administering to a subject an effective amount of a fusion protein of  claim 13 . 
     
     
         27 . The method of  claim 26 , wherein the fusion protein of  claim 13  is administered parenterally. 
     
     
         28 . A method of modulating an immune response comprising administering a fusion protein of  claim 13  in an amount sufficient to elicit a tolerogenic response. 
     
     
         29 . The method of  claim 28 , wherein the immune response is antigen-specific. 
     
     
         30 . The method of  claim 28 , wherein said administering step is carried out in vivo. 
     
     
         31 . The method of  claim 28 , wherein said administering step is carried out ex vivo. 
     
     
         32 . The method of  claim 28 , wherein the tolerogenic response is an active tolerance mechanism. 
     
     
         33 . The method of  claim 28 , wherein the tolerogenic response is a sustained tolerogenic response. 
     
     
         34 . A kit comprising one or more containers having pharmaceutical dosage units comprising an effective amount of a fusion protein of  claim 13 , wherein the container is packaged with optional instructions for the use thereof. 
     
     
         35 . A method of modulating antigen-presenting cell function comprising exposing an antigen-presenting cell to a fusion protein of  claim 13 .