Subsets of human CD4+ regulatory T cells express the peripheral homing receptor CXCR3

Regulatory T cells (Tregs) migrate into peripheral sites of inflammation such as allografts undergoing rejection, where they serve to suppress the immune response. In this study, we find that ∼30–40% of human CD25hi FOXP3+ CD4+ Tregs express the peripheral CXC chemokine receptor 3 (CXCR3) and that this subset has potent immunoregulatory properties. Consistently, we observed that proliferative responses as well as IFN‐γ production were significantly higher using CXCR3‐depleted versus undepleted responders in the mixed lymphocyte reaction, as well as following mitogen‐dependent activation of T cells. Using microfluidics, we also found that CXCR3 was functional on CXCR3pos Tregs, in as much as chemotaxis and directional persistence towards interferon‐γ‐inducible protein of 10 kDa (IP‐10) was significantly greater for CXCR3pos than CXCR3neg Tregs. Following activation, CXCR3‐expressing CD4+ Tregs were maintained in vitro in cell culture in the presence of the mammalian target of rapamycin (mTOR) inhibitor rapamycin, and we detected higher numbers of circulating CXCR3+ FOXP3+ T cells in adult and pediatric recipients of renal transplants who were treated with mTOR‐inhibitor immunosuppressive therapy. Collectively, these results demonstrate that the peripheral homing receptor CXCR3 is expressed on subset(s) of circulating human Tregs and suggest a role for CXCR3 in their recruitment into peripheral sites of inflammation.

[1]  T. Issekutz,et al.  CXCR3 blockade inhibits T‐cell migration into the CNS during EAE and prevents development of adoptively transferred, but not actively induced, disease , 2010, European journal of immunology.

[2]  Mehmet Toner,et al.  Burn Injury Reduces Neutrophil Directional Migration Speed in Microfluidic Devices , 2010, PloS one.

[3]  D. Hafler,et al.  FOXP3+ regulatory T cells in the human immune system , 2010, Nature Reviews Immunology.

[4]  K. Wood,et al.  In vivo Prevention of Transplant Arteriosclerosis by ex vivo Expanded Human Regulatory T Cells , 2010, Nature Medicine.

[5]  C. Weston,et al.  Distinct Roles for CCR4 and CXCR3 in the Recruitment and Positioning of Regulatory T Cells in the Inflamed Human Liver , 2010, The Journal of Immunology.

[6]  V. Oliveira,et al.  Alpha-1,2-Mannosidase and Hence N-Glycosylation Are Required for Regulatory T Cell Migration and Allograft Tolerance in Mice , 2010, PloS one.

[7]  Hélène Dumortier,et al.  CXCR3, Inflammation, and Autoimmune Diseases , 2009, Annals of the New York Academy of Sciences.

[8]  B. Aizenstein,et al.  Noninvasive Detection of Acute and Chronic Injuries in Human Renal Transplant by Elevation of Multiple Cytokines/Chemokines in Urine , 2009, Transplantation.

[9]  P. Worley,et al.  The mTOR kinase differentially regulates effector and regulatory T cell lineage commitment. , 2009, Immunity.

[10]  L. Bradley.  Faculty Opinions recommendation of The transcription factor T-bet controls regulatory T cell homeostasis and function during type 1 inflammation. , 2009 .

[11]  E. Shevach.  Mechanisms of foxp3+ T regulatory cell-mediated suppression. , 2009, Immunity.

[12]  J. Grinyó,et al.  Inhibitors of the Mammalian Target of Rapamycin and Transplant Tolerance , 2009, Transplantation.

[13]  R. Jessberger,et al.  Regulatory T cells sequentially migrate from inflamed tissues to draining lymph nodes to suppress the alloimmune response. , 2009, Immunity.

[14]  X. Hong,et al.  Unaltered Graft Survival and Intragraft Lymphocytes Infiltration in the Cardiac Allograft of Cxcr3−/‐ Mouse Recipients , 2008, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[15]  Qizhi Tang,et al.  The Foxp3+ regulatory T cell: a jack of all trades, master of regulation , 2008, Nature Immunology.

[16]  T. Miyazaki,et al.  Therapeutic effect of CXCR3-expressing regulatory T cells on liver, lung and intestinal damages in a murine acute GVHD model , 2008, Gene Therapy.

[17]  S. Ishihara,et al.  CD4+CD25+Foxp3+ regulatory T cells induce cytokine deprivation–mediated apoptosis of effector CD4+ T cells , 2007, Nature Immunology.

[18]  K. Boyd,et al.  The inhibitory cytokine IL-35 contributes to regulatory T-cell function , 2007, Nature.

[19]  I. Campbell,et al.  CXCR3 Signaling Reduces the Severity of Experimental Autoimmune Encephalomyelitis by Controlling the Parenchymal Distribution of Effector and Regulatory T Cells in the Central Nervous System1 , 2007, The Journal of Immunology.

[20]  M. Battaglia,et al.  Regulatory T-cell immunotherapy for tolerance to self antigens and alloantigens in humans , 2007, Nature Reviews Immunology.

[21]  V. Kuchroo,et al.  Adenosine generation catalyzed by CD39 and CD73 expressed on regulatory T cells mediates immune suppression , 2007, The Journal of experimental medicine.

[22]  V. Kuchroo,et al.  Contrasting Effects of Cyclosporine and Rapamycin in De Novo Generation of Alloantigen‐Specific Regulatory T Cells , 2007, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[23]  R. Clark,et al.  The Majority of Human Peripheral Blood CD4+CD25highFoxp3+ Regulatory T Cells Bear Functional Skin-Homing Receptors1 , 2006, The Journal of Immunology.

[24]  E. Shevach,et al.  The lifestyle of naturally occurring CD4+CD25+Foxp3+ regulatory T cells , 2006, Immunological reviews.

[25]  S. Curbishley,et al.  Epithelial Inflammation Is Associated with CCL28 Production and the Recruitment of Regulatory T Cells Expressing CCR101 , 2006, The Journal of Immunology.

[26]  J. Ingelfinger,et al.  Safety and efficacy of a calcineurin inhibitor avoidance regimen in pediatric renal transplantation. , 2006, Journal of the American Society of Nephrology : JASN.

[27]  B. Banas,et al.  Expression of the chemokine receptor CXCR3 in human renal allografts--a prospective study. , 2006, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[28]  V. Sharma,et al.  Messenger RNA for FOXP3 in the urine of renal-allograft recipients. , 2005, The New England journal of medicine.

[29]  A. Hamann,et al.  Homing to suppress: address codes for Treg migration. , 2005, Trends in immunology.

[30]  M. Dorf,et al.  Recruitment of Foxp3+ T regulatory cells mediating allograft tolerance depends on the CCR4 chemokine receptor , 2005, The Journal of experimental medicine.

[31]  R. Coffman,et al.  T cells that cannot respond to TGF-β escape control by CD4+CD25+ regulatory T cells , 2005, The Journal of experimental medicine.

[32]  F. Thaiss,et al.  CXCR3 and CCR5 Positive T-Cell Recruitment in Acute Human Renal Allograft Rejection , 2004, Transplantation.

[33]  R. Andreesen,et al.  Large scale in vitro expansion of polyclonal human CD4+CD25high regulatory T cells , 2004, Cancer Cell International.

[34]  V. Sharma,et al.  Noninvasive detection of renal allograft inflammation by measurements of mRNA for IP-10 and CXCR3 in urine. , 2004, Kidney international.

[35]  B. Aizenstein,et al.  Elevation of CXCR3‐Binding Chemokines in Urine Indicates Acute Renal‐Allograft Dysfunction , 2004, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[36]  J. Buer,et al.  Developmental Stage, Phenotype, and Migration Distinguish Naive- and Effector/Memory-like CD4+ Regulatory T Cells , 2004, The Journal of experimental medicine.

[37]  R. Flavell,et al.  CD4+CD25+ T regulatory cells control anti-islet CD8+ T cells through TGF-β–TGF-β receptor interactions in type 1 diabetes , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[38]  P. McCarthy,et al.  Chemokine and receptor-gene expression during early and late acute rejection episodes in human cardiac allografts , 2003, Transplantation.

[39]  A. Rudensky,et al.  Foxp3 programs the development and function of CD4+CD25+ regulatory T cells , 2003, Nature Immunology.

[40]  P. Libby,et al.  Differential Expression of the IFN-γ-Inducible CXCR3-Binding Chemokines, IFN-Inducible Protein 10, Monokine Induced by IFN, and IFN-Inducible T Cell α Chemoattractant in Human Cardiac Allografts: Association with Cardiac Allograft Vasculopathy and Acute Rejection1 , 2002, The Journal of Immunology.

[41]  G. McMahon,et al.  Expression of the Chemokine Receptor CXCR3 and Its Ligand IP-10 During Human Cardiac Allograft Rejection , 2001, Circulation.

[42]  G. Freeman,et al.  CD4+CD25high Regulatory Cells in Human Peripheral Blood1 , 2001, The Journal of Immunology.

[43]  A. Luster,et al.  Donor-Derived Ip-10 Initiates Development of Acute Allograft Rejection , 2001, The Journal of experimental medicine.

[44]  F. Powrie,et al.  IL-10 Is Required for Regulatory T Cells to Mediate Tolerance to Alloantigens In Vivo1 , 2001, The Journal of Immunology.

[45]  W. Hancock,et al.  Requirement of the Chemokine Receptor CXCR3 for Acute Allograft Rejection , 2000, The Journal of experimental medicine.

[46]  Fiona Powrie,et al.  Cytotoxic T Lymphocyte–Associated Antigen 4 Plays an Essential Role in the Function of Cd25+Cd4+ Regulatory Cells That Control Intestinal Inflammation , 2000, The Journal of experimental medicine.

[47]  Fiona Powrie,et al.  An Essential Role for Interleukin 10 in the Function of Regulatory T Cells That Inhibit Intestinal Inflammation , 1999, The Journal of experimental medicine.

[48]  E. Shevach,et al.  CD4+CD25+ Immunoregulatory T Cells Suppress Polyclonal T Cell Activation In Vitro by Inhibiting Interleukin 2 Production , 1998, The Journal of experimental medicine.

[49]  M. Baggiolini.  Chemokines and leukocyte traffic , 1998, Nature.

[50]  R. Goto,et al.  Vascular Biology, Atherosclerosis and Biology CD4 Regulatory T Cells Generated in Vitro with IFN- (cid:1) and Allogeneic APC Inhibit Transplant Arteriosclerosis , 2010 .

[51]  R. Negrin,et al.  Differential impact of mammalian target of rapamycin inhibition on CD4+CD25+Foxp3+ regulatory T cells compared with conventional CD4+ T cells. , 2008, Blood.

[52]  A. Rudensky,et al.  Foxp 3 programs the development and function of CD 4 + CD 25 + regulatory T cells , 2003 .

[53]  J. Casanova,et al.  X-linked neonatal diabetes mellitus, enteropathy and endocrinopathy syndrome is the human equivalent of mouse scurfy , 2001, Nature Genetics.

[54]  H. Ochs,et al.  The immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) is caused by mutations of FOXP3 , 2001, Nature Genetics.