Methods and Materials for the Generation of Regulatory T Cells

Abstract

Methods are disclosed for the generation of immunosuppressive regulatory T cells. The methods can include contacting a population of CD4+CD25− T cells with a T cell receptor (TCR)/CD3 activator, a TCR co-stimulator activator, and rapamycin. Kits for the generation of immunosuppressive regulatory T cells, methods of use, and cell populations are also disclosed.

1 . A method of generating immunosuppressive regulatory T cells from a sample, the method comprising:
 providing an initial sample containing a population of CD4+CD25− T cells; and   contacting the population with a T cell receptor (TCR)/CD3 activator, a TCR co-stimulator activator, and rapamycin to generate a final sample comprising immunosuppressive regulatory T cells.   
     
     
         2 . The method of  claim 1 , further comprising purifying the population prior to the contacting step. 
     
     
         3 . The method of  claim 2 , wherein the purifying step comprises isolating CD4+ T cells. 
     
     
         4 . The method of  claim 1 , wherein the generated immunosuppressive regulatory T cells are CD4+CD25+. 
     
     
         5 . The method of  claim 1 , wherein the generated immunosuppressive regulatory T cells are CD4+CD25+FOXP3−. 
     
     
         6 .- 8 . (canceled) 
     
     
         9 . The method of  claim 1 , wherein the final sample comprises at least about 50% CD4+CD25+ cells. 
     
     
         10 . The method of  claim 1 , wherein the population is contacted with the rapamycin before the T cell receptor (TCR)/CD3 activator and TCR co-stimulator activator. 
     
     
         11 .- 16 . (canceled) 
     
     
         17 . The method of  claim 1 , wherein the T cell receptor (TCR)/CD3 activator is an antibody or a ligand for TCR/CD3. 
     
     
         18 .- 19 . (canceled) 
     
     
         20 . The method of  claim 1 , the TCR co-stimulator activator is an antibody. 
     
     
         21 .- 36 . (canceled) 
     
     
         37 . A kit for the generation of immunosuppressive regulatory T cells, the kit comprising: a T cell receptor (TCR)/CD3 activator, a TCR co-stimulator activator, and rapamycin. 
     
     
         38 . The kit of  claim 37 , wherein:
 the T cell receptor (TCR)/CD3 activator is immobilized on a solid phase; and   the TCR co-stimulator activator is immobilized on a solid phase.   
     
     
         39 . (canceled) 
     
     
         40 . A composition comprising a population of immunosuppressive regulatory T cells, wherein at least 80% of the immunosuppressive regulatory T cells are CD4+FoxP3− T cells. 
     
     
         41 . The composition of  claim 40 , wherein the population of immunosuppressive regulatory T cells contains CD4+CD25+ T cells. 
     
     
         42 . The composition of  claim 40 , wherein the population of immunosuppressive regulatory T cells contains at least 50% CD4+CD25+ T cells. 
     
     
         43 . The composition of  claim 40 , wherein the population of immunosuppressive regulatory T cells contains CD4+CD25− T cells. 
     
     
         44 . The composition of  claim 40 , wherein the population of immunosuppressive regulatory T cells is present in a culture medium. 
     
     
         45 . A method of treating a mammal, the method comprising administering the composition of  claim 40  to the mammal.