Compositions and methods for in vitro cultivation and/or expansion of regulatory t cells

Abstract

FOXP3 + regulatory T cells (Tregs) can represent powerful adoptive immunotherapies for autoimmune diseases, metabolic diseases, and other chronic inflammatory diseases. The present invention is related to the ability to maintain and expand stable Treg lines and can provide insight into FOXP3 + Treg physiology and can enable feasible strategies of Treg-based immunotherapy.

That which is claimed: 
     
         1 . A regulatory T-cell (Treg) population that expresses a phenotypic marker and maintains the phenotypic marker for at least four weeks in culture. 
     
     
         2 . The Treg population of  claim 1 , wherein the Treg population comprises FOXP3+ Tregs and the FOXP3+ phenotype is maintained for at least four weeks in culture. 
     
     
         3 . The Treg population of  claim 1 , wherein the Treg population comprises antigen-specific FOXP3+ Tregs and the antigen-specific FOXP3+ phenotype is maintained for at least four weeks in culture. 
     
     
         4 . The Treg population of  claim 1 , wherein the Treg population comprises antigen-specific FOXP3+ Tregs and the antigen-specific FOXP3+ phenotype is maintained for at least four weeks in culture, and wherein the antigen-specificity is relevant to an autoimmune disease. 
     
     
         5 . The Treg population of  claim 1 , wherein the Treg population comprises antigen-specific FOXP3+ Tregs and the antigen-specific FOXP3+ phenotype is maintained for at least four weeks in culture, and wherein the antigen-specificity is relevant to multiple sclerosis. 
     
     
         6 . The Treg population of  claim 1 , wherein the Treg population comprises CD4+ FOXP3+ Tregs and the CD4+ FOXP3+ phenotype is maintained for at least four weeks in culture. 
     
     
         7 . The Treg population of  claim 1 , wherein the Treg population maintains a FOXP3 high  phenotype for at least four weeks in culture. 
     
     
         8 . The Treg population of  claim 1 , wherein the Treg population maintains a CD25 high  phenotype for at least four weeks in culture. 
     
     
         9 . The Treg population of  claim 1 , wherein the Treg population comprises primary, native Treg cells. 
     
     
         10 . The Treg population of  claim 1 , wherein, after four weeks in culture, at least 90% of the Tregs in the Treg population maintain the phenotypic marker. 
     
     
         11 . The Treg population of  claim 1 , wherein the Tregs in the Treg population maintain the phenotypic marker for at least eight weeks in culture. 
     
     
         12 . A pharmaceutical formulation comprising the Treg population of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         13 . A method of treating, modulating, and/or preventing an immunological disorder in a subject in need thereof comprising a step of administering a therapeutic amount of the Treg population of  claim 1  to the subject. 
     
     
         14 . An adoptive immunotherapy method comprising a step of administering a therapeutic amount of the Treg population of  claim 1  to a subject in need thereof 
     
     
         15 . A method of eliciting a tolerogenic response in a subject in need thereof comprising administering a therapeutic amount of the Treg population of  claim 1  to the subject in an amount sufficient to elicit a tolerogenic response. 
     
     
         16 . An in vitro method of preparing a regulatory T-cell (Treg) population having an antigen-specificity relevant to an autoimmune disease comprising:
 exposing T-cells to a medium comprising an anti-inflammatory cytokine and an anti-inflammatory cytokine receptor inhibitor, wherein the anti-inflammatory cytokine is IL-2 and the anti-inflammatory cytokine receptor inhibitor is an anti-CD25 agent;   propagating the T-cells in the medium comprising the anti-inflammatory cytokine and the anti-inflammatory cytokine receptor inhibitor to provide a Treg population; and   activating the Treg population in the presence of i) TGF-β and/or IFN-β and ii) an antigen, a superantigen, and/or a mitogenic stimulus to provide the Treg population having antigen-specificity relevant to an autoimmune disease.   
     
     
         17 . The method of  claim 16 , wherein the anti-inflammatory cytokine receptor inhibitor is an anti-CD25 antibody and/or a fragment thereof. 
     
     
         18 . The method of  claim 16 , wherein the Treg population maintains the antigen-specific phenotype for at least four weeks in culture. 
     
     
         19 . The method of  claim 16 , wherein, after four weeks in culture, at least 90% of the Tregs in the Treg population maintain the antigen-specific phenotype. 
     
     
         20 . A regulatory T-cell (Treg) population prepared by an in vitro method comprising:
 exposing T-cells to a medium comprising an anti-inflammatory cytokine and an anti-inflammatory cytokine receptor inhibitor; and   propagating the T-cells in the medium comprising the anti-inflammatory cytokine and the anti-inflammatory cytokine receptor inhibitor to provide the Treg population.