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 . The method of  claim 1 , wherein the population of CD4+CD25− T cells comprise CD4+CD25−CD8+ T cells. 
     
     
         7 . The method of  claim 1 , wherein the sample is blood. 
     
     
         8 . (canceled) 
     
     
         9 . The method of  claim 1 , wherein the final sample comprises at least about 50% CD4+CD25+ cells. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 1 , wherein the population is contacted with the rapamycin concurrently with the T cell receptor (TCR)/CD3 activator and TCR co-stimulator activator. 
     
     
         12 . The method of  claim 1 , wherein the rapamycin is added to the population at a concentration of about 0.01 μM to about 10 μM. 
     
     
         13 .- 14 . (canceled) 
     
     
         14 . The method of  claim 1 , wherein the contacting step further comprises contacting the population with at least one cytokine or growth factor. 
     
     
         15 . (canceled) 
     
     
         16 . The method of  claim 1 , wherein the T cell receptor (TCR)/CD3 activator is an antibody or a ligand for TCR/CD3. 
     
     
         17 .- 19 . (canceled) 
     
     
         18 . The method of  claim 1 , wherein the TCR co-stimulator activator is an antibody. 
     
     
         19 .- 22 . (canceled) 
     
     
         20 . The method of  claim 1 , wherein the TCR co-stimulator activator is a CD28 antibody. 
     
     
         21 . (canceled) 
     
     
         22 . The method of  claim 1 , 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.   
     
     
         23 . The method of  claim 22 , wherein the T cell receptor (TCR)/CD3 activator and the TCR co-stimulator activator are immobilized on the same solid phase. 
     
     
         24 . (canceled) 
     
     
         25 . The method of  claim 1 , wherein:
 the T cell receptor (TCR)/CD3 activator is immobilized on beads; and   the TCR co-stimulator activator is immobilized on beads.   
     
     
         26 . The method of  claim 22 , wherein the solid phase comprises glass, silica, latex, polymeric materials, plastic, tissue culture plastic, dextran, cellulose, polyethylene glycol, iron, or combinations thereof. 
     
     
         27 . The method of  claim 22 , wherein the solid phase comprises particles, beads, bottles, tubes, strips plates or wells, sheets, fibres, capillaries, needles, combs, pipette tips, microarrays, chips, filters, or membranes. 
     
     
         28 .- 36 . (canceled) 
     
     
         29 . 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. 
     
     
         30 .- 39 . (canceled)