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.

1 - 10 . (canceled) 
     
     
         11 . A method comprising: a) obtaining a population of T cells from an individual with asthma; b) isolating and purifying from the T cell population a subpopulation of CD4+CD25+ Tregs; and c) expanding the Treg cells of the subpopulation in the presence of effective amounts of a first and a second activator and a co-stimulator activator, wherein the expanded Treg cells exhibit enhanced suppressive activity compared to a population of freshly purified, unexpanded Tregs from the individual. 
     
     
         12 . The method of  claim 11 , wherein the expansion is between about 100 to 1,000 fold. 
     
     
         13 . The method of  claim 11 , wherein the expansion was carried out for a period of about 3 weeks. 
     
     
         14 . The method of  claim 11 , wherein the first activator is an anti-CD3 antibody. 
     
     
         15 . The method of  claim 11 , wherein the second activator is IL-2. 
     
     
         16 . The method of  claim 11 , wherein the co-stimulator activator is CD28. 
     
     
         17 . The method of  claim 11 , wherein the first activator is an anti-CD3 antibody, the second activator is IL-2 and the co-stimulator activator is CD28. 
     
     
         18 . The method of  claim 11 , wherein the isolating and purifying are carried out until the subpopulation of CD4+CD25+, Tregs is greater than 40% positive for Foxp3 and greater than 90% for CD4. 
     
     
         19 . A method of treating an individual with immune-mediated disease, the method comprising: a) obtaining a population of T cells from an individual with asthma; b) isolating and purifying from the T cell population a subpopulation of CD4+CD25+ Tregs; c) expanding the Treg cells of the subpopulation in the presence of effective amounts of a first and a second activator and a co-stimulator; and d) administering a portion of the CD4+CD25+ regulatory T cells to a human being treated for the immune-mediated disease. 
     
     
         20 . The method of  claim 19 , wherein the first activator is an anti-CD3 antibody. 
     
     
         21 . The method of  claim 19 , wherein the second activator is IL-2. 
     
     
         22 . The method of  claim 19 , wherein the co-stimulator activator is CD28. 
     
     
         23 . The method of  claim 19 , wherein the first activator is an anti-CD3 antibody, the second activator is IL-2 and the co-stimulator activator is CD28. 
     
     
         24 . The method of  claim 19 , wherein the isolating and purifying are carried out until the subpopulation of CD4+CD25, Tregs is greater than 40% positive for Foxp3 and greater than 90% for CD4.