Use of PI3K M-TOR and Akt Inhibitors to Induce FOXP3 Expression and Generate Regulatory T Cells

Abstract

The invention relates to a method of inducing Foxp3 expression in a T cell comprising (i) stimulating a T cell (ii) inhibiting signalling via PBK alpha or PBK delta or m-TOR or Akt in said T cell, wherein said inhibition is commenced 10 to 22 hours after the stimulation of (i). The invention also relates to certain uses of PBK inhibitors, PBK inhibitors for particular uses, and kits.

1 . A method of inducing Foxp3 expression in a T cell comprising
 (i) stimulating a T cell   (ii) inhibiting signalling via PI3K alpha or PI3K delta or m-TOR or Akt in said T cell, wherein said inhibition is commenced 10 to 22 hours after the stimulation of (i).   
   
   
       2 . A method of inducing Foxp3 expression in a previously stimulated T cell comprising inhibiting signalling in said T cell via PI3K alpha or PI3K delta or m-TOR or Akt, wherein said inhibition is commenced 10 to 22 hours after the stimulation. 
   
   
       3 . A method of treating a subject in need of regulatory T cell(s) comprising
 (i) removing a sample comprising a T cell from a subject   (ii) stimulating said T cell   (iii) optionally withdrawing said stimulation   (iv) inhibiting signalling via PI3K alpha or PI3K delta or m-TOR or Akt in said T cell, wherein said inhibition is commenced 10 to 22 hours after the stimulation of (i); and   (v) reintroducing said T cell to said subject.   
   
   
       4 . A method according to  claim 1  wherein said inhibition is commenced about 17 to 19 hours after the stimulation. 
   
   
       5 . A method according to  claim 4  wherein said inhibition is commenced about 18 hours after the stimulation. 
   
   
       6 . A method according to  claim 1  wherein the inhibiting signalling step comprises inhibiting signalling via PI3K alpha or PI3K delta. 
   
   
       7 . A method according to  claim 6  wherein inhibiting signalling via PI3K alpha or PI3K delta comprises contacting said cell with PI3K inhibitor, and wherein said inhibitor inhibits PI3K alpha and/or PI3K delta. 
   
   
       8 . A method according to  claim 1  further comprising withdrawing said stimulation no later than at the time of inhibiting signalling. 
   
   
       9 . A method according to  claim 1  wherein stimulating said T cell comprises stimulating said T cell via the T cell receptor (TCR). 
   
   
       10 . A method of inducing of a T cell into a regulatory T cell comprising contacting said T cell with a PI3K inhibitor wherein said inhibitor inhibits PI3K alpha and/or PI3K delta. 
   
   
       11 - 14 . (canceled) 
   
   
       15 . A kit comprising
 (i) a TCR stimulant; and   (ii) an inhibitor of PI3K alpha or delta.   
   
   
       16 . A kit according to  claim 15  wherein said TCR stimulant comprises anti-TCR and anti-CD28 antibody. 
   
   
       17 . A kit according to  claim 15  further comprising an m-TOR inhibitor. 
   
   
       18 . A kit according to  claim 17  wherein said m-TOR inhibitor is rapamycin. 
   
   
       19 . A kit according to  claim 15  further comprising an Akt inhibitor. 
   
   
       20 . A method for generating a regulatory T-cell comprising
 (i) stimulating a T cell   (ii) inhibiting signalling via PI3K alpha or PI3K delta or m-TOR or Akt in said T cell, wherein said inhibition is commenced 10 to 22 hours after the stimulation of (i).   
   
   
       21 . A method for generating a regulatory T-cell comprising treating a stimulated CD8− T cell with phosphatidyl inositol 3 kinase (PI3K) inhibitor wherein said inhibitor inhibits PI3K alpha and/or PI3K delta. 
   
   
       22 . A method according to  claim 1  wherein said PI3K inhibitor comprises LY294002. 
   
   
       23 . A method according to  claim 20  wherein said PI3K inhibitor comprises LY294002. 
   
   
       24 . A method according to  claim 10  wherein said PI3K inhibitor comprises LY294002. 
   
   
       25 . A kit according to  claim 15  wherein said PI3K inhibitor comprises LY294002.