title: Therapeutic evaluation of ex vivo-generated versus natural regulatory T-cells in a mouse model of chronic gut inflammation.
abstract: The objectives of this study were to (a) evaluate and compare the ability of ex vivo-generated induced regulatory T cells (iTregs) and freshly isolated natural Tregs (nTregs) to reverse/attenuate preexisting intestinal inflammation in a mouse model of chronic colitis and (b) quantify the Treg-targeted gene expression profiles of these two Treg populations. We found that ex vivo-generated iTregs were significantly more potent than nTregs at attenuating preexisting colitis. This superior therapeutic activity was associated with increased accumulation of iTregs within the mesenteric lymph nodes and large and significant reductions in interleukin (IL)-6 and IL-17A expression in the colons of iTreg- versus nTreg-treated mice. The enhanced immunosuppressive activity of iTregs was not because of increased expression or stability of Foxp3 as iTregs and nTregs obtained from the mesenteric lymph nodes, and colons of reconstituted mice expressed similar levels of this important transcription factor. In addition, we observed a total of 27 genes that were either upregulated or downregulated in iTregs when compared with nTregs. Although iTregs were found to be superior at reversing established disease, their message levels of IL-10 and IL-35 and surface expression of the gut-homing molecules CCR9 and α4β7 were significantly reduced when compared with nTregs. Taken together, our data demonstrate that ex vivo-generated iTregs are significantly more potent than nTregs at attenuating preexisting gut inflammation despite reduced expression of classical regulatory cytokines and gut-homing molecules. Our data suggest that the immunosuppressive activity of iTregs may be because of their ability to directly or indirectly decrease expression of IL-6 and IL-17A within the inflamed bowel.
authors.affiliation: *Department of Molecular and Cellular Physiology, †Department of Microbiology and Immunology, and ‡Department of Pathology, Louisiana State University Health Sciences Center, Shreveport, Louisiana; and §Department of Immunology and Molecular Microbiology, Texas Tech University Health Sciences Center, Lubbock, Texas.
authors.forename: Fridrik
authors.initials: F
authors.lastname: Karlsson
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Martinez
authors.forename: Laura
authors.initials: L
authors.lastname: Gray
authors.forename: Songlin
authors.initials: S
authors.lastname: Zhang
authors.forename: Ikuo
authors.initials: I
authors.lastname: Tsunoda
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Grisham
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002908:Chronic Disease; D003092:Colitis / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D051858:Forkhead Transcription Factors / Q000502:physiology*; D041981:Gastrointestinal Tract / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D018398:Homeodomain Proteins / Q000502:physiology*; D006801:Humans; D007249:Inflammation / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology* / Q000378:metabolism
chemical_list: D016207:Cytokines; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D018398:Homeodomain Proteins; C064658:RAG-1 protein
references.citation: J Immunol. 2003 Nov 15;171(10):5018-26
references.pmid: 14607898
references.citation: Science. 2010 Sep 24;329(5999):1667-71
references.pmid: 20929851
references.citation: J Immunol. 2009 Mar 15;182(6):3461-8
references.pmid: 19265124
references.citation: J Exp Med. 2007 Aug 6;204(8):1765-74
references.pmid: 17620363
references.citation: Annu Rev Immunol. 2009;27:485-517
references.pmid: 19132915
references.citation: J Exp Med. 2003 Dec 15;198(12):1875-86
references.pmid: 14676299
references.citation: J Immunol. 2010 Mar 1;184(5):2458-67
references.pmid: 20118278
references.citation: Immunol Rev. 2006 Aug;212:217-37
references.pmid: 16903917
references.citation: Nat Rev Immunol. 2008 Jul;8(7):523-32
references.pmid: 18566595
references.citation: Annu Rev Immunol. 2012;30:531-64
references.pmid: 22224781
references.citation: J Clin Invest. 2007 Mar;117(3):514-21
references.pmid: 17332878
references.citation: Annu Rev Immunol. 2012;30:733-58
references.pmid: 22224762
references.citation: Nat Immunol. 2009 Sep;10(9):1000-7
references.pmid: 19633673
references.citation: Nature. 2012 Feb 08;482(7385):395-9
references.pmid: 22318520
references.citation: Immunol Rev. 2009 Jul;230(1):144-59
references.pmid: 19594634
references.citation: Arthritis Rheum. 2012 Aug;64(8):2548-58
references.pmid: 22605463
references.citation: Immunity. 2008 Jul 18;29(1):44-56
references.pmid: 18585065
references.citation: Sci Transl Med. 2011 May 18;3(83):83ra41
references.pmid: 21593401
references.citation: J Exp Med. 2004 Jun 7;199(11):1455-65
references.pmid: 15184499
references.citation: Immunol Rev. 2006 Aug;212:8-27
references.pmid: 16903903
references.citation: Blood. 2007 Oct 1;110(7):2501-10
references.pmid: 17575071
references.citation: PLoS One. 2012;7(3):e34547
references.pmid: 22479644
references.citation: Blood. 2007 Mar 1;109(5):2058-65
references.pmid: 17110462
references.citation: Am J Physiol Gastrointest Liver Physiol. 2009 Feb;296(2):G135-46
references.pmid: 19033538
references.citation: J Immunol. 2010 Sep 1;185(5):2675-9
references.pmid: 20679534
references.citation: J Immunol. 2007 Sep 15;179(6):3724-33
references.pmid: 17785809
references.citation: World J Gastroenterol. 2011 Feb 7;17(5):578-93
references.pmid: 21350707
references.citation: J Exp Med. 2009 Nov 23;206(12):2701-15
references.pmid: 19917773
references.citation: J Immunol. 2001 Mar 1;166(5):3008-18
references.pmid: 11207250
references.citation: BMC Bioinformatics. 2006 Feb 22;7:85
references.pmid: 16504059
references.citation: J Immunol. 2010 Sep 15;185(6):3295-304
references.pmid: 20720211
references.citation: J Exp Med. 2008 Sep 1;205(9):2139-49
references.pmid: 18710932
references.citation: Nat Immunol. 2007 May;8(5):457-62
references.pmid: 17440451
references.citation: J Immunol. 2011 Jun 1;186(11):6329-37
references.pmid: 21525380
references.citation: J Immunol. 2007 Jan 1;178(1):291-300
references.pmid: 17182566
references.citation: Immunol Rev. 2008 Jun;223:371-90
references.pmid: 18613848
references.citation: Nat Protoc. 2007;2(7):1789-94
references.pmid: 17641646
references.citation: Immunol Rev. 2006 Aug;212:256-71
references.pmid: 16903919
references.citation: J Immunol. 2007 Jun 1;178(11):6725-9
references.pmid: 17513718
references.citation: J Immunol. 2009 Jul 15;183(2):975-83
references.pmid: 19542365
references.citation: Eur J Immunol. 2010 Oct;40(10):2902-13
references.pmid: 20806292
references.citation: J Immunol. 2005 Jul 15;175(2):944-50
references.pmid: 16002693
references.citation: J Immunol. 2011 Jun 15;186(12):6779-87
references.pmid: 21572024
references.citation: Nat Immunol. 2007 Dec;8(12):1285-7
references.pmid: 18026078
references.citation: J Exp Med. 2003 Feb 17;197(4):403-11
references.pmid: 12591899
references.citation: Inflamm Bowel Dis. 2008 Feb;14(2):275-89
references.pmid: 17924560
references.citation: J Immunol. 2008 Dec 15;181(12):8209-13
references.pmid: 19050237
references.citation: Inflamm Bowel Dis. 2011 May;17(5):1229-45
references.pmid: 21312318
references.citation: Transplant Proc. 2009 Jul-Aug;41(6):2670-2
references.pmid: 19715998
references.citation: J Immunol. 2010 May 15;184(10):5519-26
references.pmid: 20400707
references.citation: J Immunol. 2003 Apr 15;170(8):3939-43
references.pmid: 12682220
references.citation: Science. 2007 Jul 13;317(5835):256-60
references.pmid: 17569825
references.citation: J Mol Cell Biol. 2012 Feb;4(1):22-8
references.pmid: 22107826
references.citation: Methods Mol Biol. 2011;677:47-61
references.pmid: 20941602
references.citation: Mol Immunol. 2009 Jul;46(11-12):2424-8
references.pmid: 19428111
references.citation: Am J Physiol Gastrointest Liver Physiol. 2006 Jan;290(1):G109-19
references.pmid: 16099868
references.citation: Gut. 2006 May;55(5):671-80
references.pmid: 16162681
references.citation: J Immunol. 2006 Nov 1;177(9):5852-60
references.pmid: 17056509
references.citation: Immunol Rev. 2011 May;241(1):260-8
references.pmid: 21488902
references.citation: J Immunol. 2011 Jan 1;186(1):32-40
references.pmid: 21106853
references.citation: Nature. 1995 Dec 14;378(6558):736-9
references.pmid: 7501023
references.citation: Immunity. 2009 Sep 18;31(3):401-11
references.pmid: 19766083
references.citation: J Immunol. 2005 Jun 15;174(12):7487-91
references.pmid: 15944246
references.citation: J Exp Med. 2007 Aug 6;204(8):1757-64
references.pmid: 17620361
references.citation: J Immunol. 2010 Apr 1;184(7):3433-41
references.pmid: 20181882
references.citation: Nature. 2011 Jun 15;474(7351):307-17
references.pmid: 21677747
references.citation: Immunity. 2008 Nov 14;29(5):758-70
references.pmid: 19006694
references.citation: Am J Transplant. 2011 Jun;11(6):1148-57
references.pmid: 21564534
references.citation: J Immunol. 2008 Jun 1;180(11):7112-6
references.pmid: 18490709
references.citation: Nature. 2007 Nov 22;450(7169):566-9
references.pmid: 18033300
references.citation: Immunity. 2005 Mar;22(3):329-41
references.pmid: 15780990
references.citation: Gastroenterology. 2011 May;140(6):1756-1767
references.pmid: 21530742
references.citation: J Immunol. 2006 Nov 15;177(10):6940-51
references.pmid: 17082609
references.citation: J Immunol. 2011 Jan 1;186(1):41-52
references.pmid: 21098236
references.citation: Am J Physiol Gastrointest Liver Physiol. 2007 Jun;292(6):G1706-14
references.pmid: 17332469
references.citation: J Exp Med. 2006 Mar 20;203(3):489-92
references.pmid: 16533888
references.citation: Eur J Immunol. 2008 Dec;38(12):3274-81
references.pmid: 19039774