IL‐10 inhibits endothelium‐dependent T cell costimulation by up‐regulation of ILT3/4 in human vascular endothelial cells

Effects of IL‐10 on endothelium‐dependent T cell activation have not been investigated in detail. We confirm expression of the IL‐10 receptor and effective signaling via STAT‐3 in human umbilical vein endothelial cells (HUVEC). In CD4 T cell cocultures with HUVEC, pretreatment of endothelial cells with IL‐10 resulted in significant dose‐dependent inhibition of CD4 T cell proliferation, which also occurred when IL‐10 was removed after pretreatment before starting cocultures. Th1/Th2 polarization of proliferated T cells, endothelial nitric oxide (NO), or IL‐12 production were unchanged. However, IL‐10 stimulation resulted in up‐regulation of SOCS‐3, a negative regulator of cytokine secretion, and induction of the inhibitory surface molecules immunoglobulin‐like transcript 3 and 4 (ILT3/ILT4) in EC, potentially involving glucocorticoid‐induced leucine zipper (GILZ). Addition of blocking antibodies against ILT3/ILT4 to EC/T cell cocultures resulted in nearly complete reestablishment of T cell proliferation. In contrast, addition of soluble ILT3 or overexpression of ILT3 in cocultures significantly reduced T cell proliferation. No induction of foxp3+ regulatory T cells was seen. In conclusion, the T cell costimulatory potential of human EC is markedly suppressed by IL‐10 due to up‐regulation of ILT3/ILT4, obviously not involving generation of Treg. This identifies a novel action of IL‐10 in EC and a potential therapeutical target for local immunomodulation.

[1]  L. Tesson,et al.  Interleukin-10 produced by recombinant adenovirus prolongs survival of cardiac allografts in rats , 2000, Gene Therapy.

[2]  M. Pallardy,et al.  GILZ expression in human dendritic cells redirects their maturation and prevents antigen-specific T lymphocyte response. , 2006, Blood.

[3]  M. Colonna,et al.  Tolerization of dendritic cells by TS cells: the crucial role of inhibitory receptors ILT3 and ILT4 , 2002, Nature Immunology.

[4]  M. Fishbein,et al.  Overexpression of Interleukin-10 by Activated T Lymphocytes Inhibits Atherosclerosis in LDL Receptor–Deficient Mice by Altering Lymphocyte and Macrophage Phenotypes , 2002, Circulation research.

[5]  J. Fan,et al.  Regulation of ILT3 Gene Expression by Processing of Precursor Transcripts in Human Endothelial Cells , 2006, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[6]  M. Territo,et al.  Interleukin-10 blocks atherosclerotic events in vitro and in vivo. , 1999, Arteriosclerosis, thrombosis, and vascular biology.

[7]  J. Pober,et al.  Human Vascular Endothelial Cells Stimulate a Lower Frequency of Alloreactive CD8+ Pre-CTL and Induce Less Clonal Expansion than Matching B Lymphoblastoid Cells: Development of a Novel Limiting Dilution Analysis Method Based on CFSE Labeling of Lymphocytes1 , 2001, The Journal of Immunology.

[8]  G. Stark,et al.  JAKs, STATs and signal transduction in response to the interferons and other cytokines. , 1996, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[9]  A. Foussat,et al.  Synthesis of glucocorticoid-induced leucine zipper (GILZ) by macrophages: an anti-inflammatory and immunosuppressive mechanism shared by glucocorticoids and IL-10. , 2003, Blood.

[10]  C. Parish,et al.  New fluorescent dyes for lymphocyte migration studies. Analysis by flow cytometry and fluorescence microscopy. , 1990, Journal of immunological methods.

[11]  Massimo Gadina,et al.  Cytokines and Transcription Factors That Regulate T Helper Cell Differentiation: New Players and New Insights , 2003, Journal of Clinical Immunology.

[12]  M. Hecker,et al.  Interleukin-10 Induction of Nitric-oxide Synthase Expression Attenuates CD40-mediated Interleukin-12 Synthesis in Human Endothelial Cells* , 2003, Journal of Biological Chemistry.

[13]  A. Rudensky,et al.  Regulatory T cell lineage specification by the forkhead transcription factor foxp3. , 2005, Immunity.

[14]  H. Dickensheets,et al.  The interleukin-10 signal transduction pathway and regulation of gene expression in mononuclear phagocytes. , 1999, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[15]  H. Nawata,et al.  TGF-β1 Plays an Important Role in the Mechanism of CD4+CD25+ Regulatory T Cell Activity in Both Humans and Mice , 2004, The Journal of Immunology.

[16]  J. Fung,et al.  Differential effects of exogenous interleukin-10 on cardiac allograft survival: inhibition of rejection by recipient pretreatment reflects impaired host accessory cell function. , 1999, Transplantation.

[17]  J. Pober,et al.  Human Vascular Endothelial Cells Stimulate Memory But Not Naive CD8+ T Cells to Differentiate into CTL Retaining an Early Activation Phenotype1 , 2000, The Journal of Immunology.

[18]  D. Hilton,et al.  SOCS Proteins: Negative Regulators of Cytokine Signaling , 2001, Stem cells.

[19]  Manfred Brockhaus,et al.  A Novel Inhibitory Receptor (ILT3) Expressed on Monocytes, Macrophages, and Dendritic Cells Involved in Antigen Processing , 1997, The Journal of experimental medicine.

[20]  A. O’Garra,et al.  IL-10-producing and naturally occurring CD4+ Tregs: limiting collateral damage. , 2004, The Journal of clinical investigation.

[21]  D. Heistad,et al.  IL-10 deficiency increases superoxide and endothelial dysfunction during inflammation. , 2000, American journal of physiology. Heart and circulatory physiology.

[22]  Richard N. Mitchell,et al.  Interleukin-10 (IL-10) augments allograft arterial disease: paradoxical effects of IL-10 in vivo. , 1999, The American journal of pathology.

[23]  J. Pober,et al.  Human endothelial cells induce and regulate cytolytic T cell differentiation. , 1998, Journal of immunology.

[24]  S. Kim-Schulze,et al.  Recombinant Ig-Like Transcript 3-Fc Modulates T Cell Responses via Induction of Th Anergy and Differentiation of CD8+ T Suppressor Cells1 , 2006, The Journal of Immunology.

[25]  Marcela V Maus,et al.  A cell-based artificial antigen-presenting cell coated with anti-CD3 and CD28 antibodies enables rapid expansion and long-term growth of CD4 T lymphocytes. , 2002, Clinical immunology.

[26]  A. Yoshimura,et al.  Interleukin-10 (IL-10) selectively enhances CIS3/SOCS3 mRNA expression in human neutrophils: evidence for an IL-10-induced pathway that is independent of STAT protein activation. , 1999, Blood.

[27]  T. Mak,et al.  Immunologic Self-Tolerance Maintained by Cd25+Cd4+Regulatory T Cells Constitutively Expressing Cytotoxic T Lymphocyte–Associated Antigen 4 , 2000, The Journal of experimental medicine.

[28]  F. Ramsdell,et al.  Foxp3 and natural regulatory T cells: key to a cell lineage? , 2003, Immunity.

[29]  D. Mancini,et al.  Alloantigen specific CD8+CD28- FOXP3+ T suppressor cells induce ILT3+ ILT4+ tolerogenic endothelial cells, inhibiting alloreactivity. , 2004, International immunology.

[30]  J. Pober,et al.  T lymphocyte-endothelial cell interactions. , 2004, Annual review of immunology.

[31]  H. Valantine.  Cardiac allograft vasculopathy: central role of endothelial injury leading to transplant “atheroma” , 2003, Transplantation.

[32]  R. Coffman,et al.  Interleukin-10 and the interleukin-10 receptor. , 2001, Annual review of immunology.

[33]  L. Crepaldi,et al.  Interleukin-10 and cAMP-elevating agents cooperate to induce suppressor of cytokine signaling-3 via a protein kinase A-independent signal. , 2002, European cytokine network.

[34]  M. Kubin,et al.  A family of human lymphoid and myeloid Ig-like receptors, some of which bind to MHC class I molecules. , 1997, Journal of immunology.

[35]  R. Paus,et al.  Interleukin-15 protects from lethal apoptosis in vivo , 1997, Nature Medicine.

[36]  W. von Scheidt,et al.  Cardiac allograft vasculopathy: a review. , 1997, Circulation.

[37]  G. Vlad,et al.  Interleukin-10 induces the upregulation of the inhibitory receptor ILT4 in monocytes from HIV positive individuals. , 2003, Human immunology.

[38]  C. Drake,et al.  Role of LAG-3 in regulatory T cells. , 2004, Immunity.

[39]  J. Pober,et al.  Human vascular endothelial cells favor clonal expansion of unusual alloreactive CTL. , 1999, Journal of immunology.

[40]  M. Caligiuri,et al.  STAT‐1‐mediated repression of monocyte interleukin‐10 gene expression in vivo , 2006, European journal of immunology.

[41]  P. Libby,et al.  Vascular cells have limited capacities to activate and differentiate T cells: implications for transplant vascular sclerosis. , 1997, Transplant immunology.

[42]  M. Carrier,et al.  The central role of the endothelium in graft coronary vasculopathy and heart transplantation. , 2005, The Canadian journal of cardiology.

[43]  M. Fishbein,et al.  Regulated interleukin-10 expression prevents chronic rejection of transplanted hearts. , 2003, The Journal of thoracic and cardiovascular surgery.

[44]  Ashok Kumar,et al.  STAT-1 Mediates the Stimulatory Effect of IL-10 on CD14 Expression in Human Monocytic Cells1 , 2005, The Journal of Immunology.

[45]  J. Johnston,et al.  Lipopolysaccharide induces in macrophages the synthesis of the suppressor of cytokine signaling 3 and suppresses signal transduction in response to the activating factor IFN-gamma. , 1999, Journal of immunology.

[46]  W. Fiers,et al.  Two distinct monokines, interleukin 1 and tumor necrosis factor, each independently induce biosynthesis and transient expression of the same antigen on the surface of cultured human vascular endothelial cells. , 1986, Journal of immunology.

[47]  C. Orosz,et al.  Can graft endothelial cells initiate a host anti-graft immune response? , 1996, Transplantation.

[48]  M. Lotze,et al.  Retroviral delivery of viral interleukin-10 into myeloid dendritic cells markedly inhibits their allostimulatory activity and promotes the induction of T-cell hyporesponsiveness. , 1998, Transplantation.

[49]  C. Martínez-A,et al.  Cell surface CD28 levels define four CD4+ T cell subsets: abnormal expression in rheumatoid arthritis. , 2001, Clinical immunology.

[50]  J. Kuiper,et al.  Attenuation of atherogenesis by systemic and local adenovirus‐mediated gene transfer of interleukin‐10 in LDLr ‐/‐ Mice , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[51]  A. Yoshimura,et al.  Involvement of Suppressor of Cytokine Signaling-3 as a Mediator of the Inhibitory Effects of IL-10 on Lipopolysaccharide-Induced Macrophage Activation1 , 2002, The Journal of Immunology.

[52]  D. Mancini,et al.  High expression of ILT3 and ILT4 is a general feature of tolerogenic dendritic cells. , 2003, Transplant immunology.

[53]  J. Miyazaki,et al.  High-level expression of viral interleukin-10 in cardiac allografts fails to prolong graft survival , 2002, Transplantation.