title: Enhancement of antigen-specific Treg vaccination in vivo.
abstract: The conversion of naive T cells into Treg can be achieved in vivo by delivery of antigen under subimmunogenic conditions. Here we have examined several drugs for their ability to enhance the conversion process in vivo and have found that the rapamycin analog everolimus potently enhances Treg conversion by interfering with T-cell costimulation, reducing cell division and thereby activation of DNA methyltransferase 1 as well as by reducing T-cell activation through the ATP-gated P2×7 receptor controlling Ca2(+) influx. The resulting Tregs exhibit increased stability of Foxp3 expression even when generated in TGFβ-containing media in vitro. Thus the mammalian target of rapamycin (mTOR) inhibitor everolimus in addition to inhibiting immune responses enhances Treg conversion by several distinct pathways. The converted Tregs can be further expanded by injection of IL-2/IL-2ab complexes. These complexes also increase the number of CD25(+)Foxp3(-) cells that, however, do not represent cytokine secreting effector cells but anergic cells, some of which can secrete IL-10 and can themselves be considered regulatory T cells as well. The combined use of everolimus and IL-2/IL-2ab complexes in vivo makes it feasible to achieve highly effective antigen-driven conversion of naive T cells into Treg and their expansion in vivo and thereby the described protocols constitute important tools to achieve immunological tolerance by Treg vaccination.
authors.affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
authors.forename: Carolin
authors.initials: C
authors.lastname: Daniel
authors.forename: Kerstin
authors.initials: K
authors.lastname: Wennhold
authors.forename: Hye-Jung
authors.initials: HJ
authors.lastname: Kim
authors.forename: Harald
authors.initials: H
authors.lastname: von Boehmer
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology*; D049109:Cell Proliferation / Q000187:drug effects; D000068338:Everolimus; D005260:Female; D051858:Forkhead Transcription Factors / Q000276:immunology; D007166:Immunosuppressive Agents / Q000494:pharmacology; D016753:Interleukin-10 / Q000276:immunology; D007376:Interleukin-2 / Q000276:immunology; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D018048:Receptors, Purinergic P2 / Q000276:immunology; D058486:Receptors, Purinergic P2X7; D020123:Sirolimus / Q000031:analogs & derivatives / Q000494:pharmacology; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D014611:Vaccination
chemical_list: D000941:Antigens; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D007166:Immunosuppressive Agents; D007376:Interleukin-2; D053645:Interleukin-2 Receptor alpha Subunit; C546469:P2rx7 protein, mouse; D018048:Receptors, Purinergic P2; D058486:Receptors, Purinergic P2X7; D016753:Interleukin-10; D000068338:Everolimus; D020123:Sirolimus
references.citation: Annu Rev Physiol. 2009;71:333-59
references.pmid: 18851707
references.citation: J Immunol. 2009 Jun 1;182(11):6648-52
references.pmid: 19454658
references.citation: Proc Natl Acad Sci U S A. 2009 Jul 21;106(29):12055-60
references.pmid: 19567830
references.citation: Blood. 2005 Jun 15;105(12):4743-8
references.pmid: 15746082
references.citation: J Exp Med. 1994 Jul 1;180(1):25-34
references.pmid: 8006585
references.citation: J Exp Med. 2010 Aug 2;207(8):1701-11
references.pmid: 20660617
references.citation: Science. 2006 Mar 31;311(5769):1924-7
references.pmid: 16484453
references.citation: J Exp Med. 2009 Apr 13;206(4):751-60
references.pmid: 19332874
references.citation: J Exp Med. 2003 Dec 15;198(12):1875-86
references.pmid: 14676299
references.citation: Immunity. 2005 Mar;22(3):329-41
references.pmid: 15780990
references.citation: Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3479-84
references.pmid: 18299571
references.citation: Proc Natl Acad Sci U S A. 2008 Jun 3;105(22):7797-802
references.pmid: 18509048
references.citation: Nat Immunol. 2005 Dec;6(12):1219-27
references.pmid: 16244650
references.citation: Int Immunol. 2005 Apr;17(4):335-42
references.pmid: 15710910
references.citation: J Exp Med. 2007 Jul 9;204(7):1543-51
references.pmid: 17591856
references.citation: J Exp Med. 2004 May 17;199(10):1401-8
references.pmid: 15148338
references.citation: J Exp Med. 2005 Aug 1;202(3):445-55
references.pmid: 16061730
references.citation: Proc Natl Acad Sci U S A. 2003 Jul 22;100(15):8886-91
references.pmid: 12857946
references.citation: Immunity. 2009 May;30(5):626-35
references.pmid: 19464985
references.citation: Immunity. 2008 Jul 18;29(1):114-26
references.pmid: 18617425
references.citation: Nat Immunol. 2002 Aug;3(8):756-63
references.pmid: 12089509
references.citation: Eur J Immunol. 2008 Jun;38(6):1654-63
references.pmid: 18493985
references.citation: Methods Mol Biol. 2011;707:173-85
references.pmid: 21287335
references.citation: PLoS One. 2008 Feb 20;3(2):e1612
references.pmid: 18286169
references.citation: Nat Immunol. 2000 Jul;1(1):54-8
references.pmid: 10881175
references.citation: Immunity. 2009 Jun 19;30(6):832-44
references.pmid: 19538929
references.citation: Physiol Rev. 2002 Oct;82(4):1013-67
references.pmid: 12270951
references.citation: J Biol Chem. 1993 Apr 15;268(11):8199-203
references.pmid: 8463330
references.citation: J Exp Med. 1963 Apr 1;117:621-31
references.pmid: 14027809
references.citation: J Exp Med. 2008 Mar 17;205(3):565-74
references.pmid: 18283119
references.citation: Eur J Immunol. 2007 Sep;37(9):2378-89
references.pmid: 17694575
references.citation: Nat Protoc. 2006;1(2):653-61
references.pmid: 17802642
references.citation: Nat Immunol. 2009 Oct;10(10):1057-63
references.pmid: 19701188