Method for ex-vivo expansion of regulatory t cells with enhanced suppressive function for clinical application in immune mediated diseases

Abstract

The invention provides methods for the ex-vivo expansion of CD4+CD25+ Tregs. The invention provides a method for producing ex vivo expanded Tregs that may be used to inhibit unwanted human immune responses against self-antigens or allergens. Additionally, the ex vivo expanded Tregs may provide treatment for inflammatory/autoimmune diseases.

What is claimed is: 
     
         1 . A method of treating an individual with immune-mediated disease, the method comprising:
 a) obtaining a population of T cells from an individual with an immune-mediated disease selected from Systemic Lupus Erythematous, Multiple Sclerosis, asthma, Rheumatoid Arthritis, Crohn's Disease, or Ulcerative Colitis;   b) isolating and purifying from the T cell population a subpopulation of CD4+CD25+ Tregs;   c) expanding the Treg cells of the subpopulation over 1,000 fold, in the presence of IL-2, anti-CD3 antibodies, and anti-CD28 antibodies; and   d) administering a portion of the CD4+CD25+ regulatory T cells to a human being treated for the immune-mediated disease.   
     
     
         2 . The method of  claim 1 , wherein the immune-mediated disease is Systemic Lupus Erythematous. 
     
     
         3 . The method of  claim 1 , wherein the immune-mediated disease is Multiple Sclerosis. 
     
     
         4 . The method of  claim 1 , wherein the immune-mediated disease is asthma, Rheumatoid Arthritis. 
     
     
         5 . The method of  claim 1 , wherein the immune-mediated disease is Rheumatoid Arthritis. 
     
     
         6 . The method of  claim 1 , wherein the immune-mediated disease is Crohn's Disease. 
     
     
         7 . The method of  claim 1 , wherein the immune-mediated disease is Ulcerative Colitis. 
     
     
         8 . The method of  claim 1 , wherein the expanded Treg cells exhibit enhanced suppressive activity compared to a population of freshly purified, unexpanded Tregs from the individual. 
     
     
         9 . The method of  claim 1 , wherein the expansion is carried out over a period of about 3 weeks. 
     
     
         10 . The method of  claim 1 , wherein the anti-CD3 antibodies and anti-CD28 antibodies are immobilized on a three-dimensional solid surface. 
     
     
         11 . The method of  claim 10 , wherein the three-dimensional solid surface is a bead. 
     
     
         12 . The method of  claim 11 , wherein a Treg to bead ratio is 1:3. 
     
     
         13 . The method of  claim 11 , the bead is about 1 to 10 microns in diameter. 
     
     
         14 . The method of  claim 11 , wherein an anti-CD3 and anti-CD28 antibody ratio on the surface of the bead is from about 1:20 to about 20:1.