Interleukin-2 muteins for the expansion of T-regulatory cells
Provided herein are IL-2 muteins and IL-2 mutein Fc-fusion molecules that preferentially expand and activate T regulatory cells and are amenable to large scale production. Also provided herein are variant human IgG1 Fc molecules lacking or with highly reduced effector function and high stability despite lacking glycosylation at N297. Also, provided herein are linker peptides that are glycosylated when expressed in mammalian cells.
1. A method of increasing the ratio of regulatory T cells (Tregs) to non-regulatory T cells in a subject with an inflammatory disease, said method comprising administering to said subject an effective amount of an IL-2 mutein comprising a V91K substitution and an amino acid sequence at least 95% identical to the amino acid sequence set forth in SEQ ID NO:1.
2. The method of claim 1 , wherein the inflammatory disease is selected from the group consisting of atopic diseases, paraneoplastic autoimmune diseases, cartilage inflammation, arthritis, rheumatoid arthritis, juvenile arthritis, juvenile rheumatoid arthritis, pauciarticular juvenile rheumatoid arthritis, polyarticular juvenile rheumatoid arthritis, systemic onset juvenile rheumatoid arthritis, juvenile ankylosing spondylitis, juvenile enteropathic arthritis, juvenile reactive arthritis, juvenile Reiter's Syndrome Seronegativity Enthesopathy Arthropathy Syndrome (SEA Syndrome), juvenile dermatomyositis, juvenile psoriatic arthritis, juvenile scleroderma, juvenile systemic lupus erythematosus, juvenile vasculitis, pauciarticular rheumatoid arthritis, polyarticular rheumatoid arthritis, systemic onset rheumatoid arthritis, ankylosing spondylitis, enteropathic arthritis, reactive arthritis, Reiter's Syndrome, dermatomyositis, psoriatic arthritis, scleroderma, vasculitis, myolitis, polymyolitis, polyarteritis nodossa, Wegener's granulomatosis, arteritis, ploymyalgia rheumatica, sarcoidosis, sclerosis, primary biliary sclerosis, sclerosing cholangitis, Sjogren's syndrome, psoriasis, plaque psoriasis, guttate psoriasis, inverse psoriasis, pustular psoriasis, erythrodermic psoriasis, dermatitis, atopic dermatitis, atherosclerosis, lupus, Still's disease, Systemic Lupus Erythematosus (SLE), myasthenia gravis, inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, celiac disease, multiple sclerosis (MS), asthma, chronic obstructive pulmonary disease (COPD), rhinosinusitis, rhinosinusitis with polyps, eosinophilic esophogitis, eosinophilic bronchitis, Guillain-Barre disease, Type I diabetes mellitus, thyroiditis, Graves' disease, Addison's disease, Raynaud's phenomenon, autoimmune hepatitis, graft-versus-host disease (GVHD), transplantation rejection, and hepatitis C-induced vasculitis.
3. The method of claim 2 , wherein the inflammatory disease is ulcerative colitis.
4. A method of increasing the ratio of regulatory T cells (Tregs) to non-regulatory T cells in a subject with an inflammatory disease, said method comprising administering to said subject an effective amount of a Fc-fusion protein comprising a Fc region and a human interleukin-2 (IL-2) mutein, wherein the mutein comprises a V91K substitution and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 1, and further wherein the Fc region comprises a substitution at N297 in Eu numbering scheme, and an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO:3, wherein said Fc-fusion protein preferentially stimulates T regulatory cells.
5. The method of claim 4 , wherein the Fc is a human IgG1 Fc.
6. The method of claim 5 , further comprising a substitution or deletion of the C-terminal lysine of said human IgG1 Fc.
7. The method of claim 4 , wherein the substitution at N297 is N297G.
8. The method of claim 4 , wherein the Fc region is on the N-terminal portion.
9. The method of claim 4 , wherein a linker connects the Fc region and human IL-2 mutein of said protein, and wherein the linker is GGGGS (SEQ ID NO: 5), GGNGT (SEQ ID NO: 6), or YGNGT (SEQ ID NO: 7).
10. The method of claim 4 , wherein the human IL-2 mutein further comprises a substitution at position 125, wherein the substitution is to an alanine or serine residue.
11. The method of claim 10 , wherein the substitution at position 125 is to alanine.
12. The method of claim 4 , wherein the amino acid sequence of said Fc-fusion protein comprises the amino acid sequence of SEQ ID NO: 17.
13. The method of claim 12 , wherein the ratio of regulatory T cells (Tregs) to non-regulatory T cells is within the peripheral blood of the subject.
14. The method of claim 4 , wherein the amino acid sequence of said Fc-fusion protein comprises the amino acid sequence of SEQ ID NO:18.
15. The method of claim 14 , wherein the ratio of regulatory T cells (Tregs) to non-regulatory T cells is within the peripheral blood of the subject.
16. The method of claim 4 , wherein the inflammatory disease is selected from the group consisting of atopic diseases, paraneoplastic autoimmune diseases, cartilage inflammation, arthritis, rheumatoid arthritis, juvenile arthritis, juvenile rheumatoid arthritis, pauciarticular juvenile rheumatoid arthritis, polyarticular juvenile rheumatoid arthritis, systemic onset juvenile rheumatoid arthritis, juvenile ankylosing spondylitis, juvenile enteropathic arthritis, juvenile reactive arthritis, juvenile Reiter's Syndrome, Seronegativity Enthesopathy Arthropathy Syndrome (SEA Syndrome), juvenile dermatomyositis, juvenile psoriatic arthritis, juvenile scleroderma, juvenile systemic lupus erythematosus, juvenile vasculitis, pauciarticular rheumatoid arthritis, polyarticular rheumatoid arthritis, systemic onset rheumatoid arthritis, ankylosing spondylitis, enteropathic arthritis, reactive arthritis, Reiter's Syndrome, dermatomyositis, psoriatic arthritis, scleroderma, vasculitis, myolitis, polymyolitis, polyarteritis nodossa, Wegener's granulomatosis, arteritis, ploymyalgia rheumatica, sarcoidosis, sclerosis, primary biliary sclerosis, sclerosing cholangitis, Sjogren's syndrome, psoriasis, plaque psoriasis, guttate psoriasis, inverse psoriasis, pustular psoriasis, erythrodermic psoriasis, dermatitis, atopic dermatitis, atherosclerosis, lupus, Still's disease, Systemic Lupus Erythematosus (SLE), myasthenia gravis, inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, celiac disease, multiple sclerosis (MS), asthma, chronic obstructive pulmonary disease (COPD), rhinosinusitis, rhinosinusitis with polyps, eosinophilic esophogitis, eosinophilic bronchitis, Guillain-Barre disease, Type I diabetes mellitus, thyroiditis, Graves' disease, Addison's disease, Raynaud's phenomenon, autoimmune hepatitis, graft-versus-host disease (GVHD), transplantation rejection, and hepatitis C-induced vasculitis.
17. The method of claim 16 , wherein the inflammatory disease is ulcerative colitis.