IL-10 inhibits CD28 and ICOS costimulations of T cells via src homology 2 domain— containing protein tyrosine phosphatase 1 Alison Taylor, PhD, Mübeccel.

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Presentation transcript:

IL-10 inhibits CD28 and ICOS costimulations of T cells via src homology 2 domain— containing protein tyrosine phosphatase 1 Alison Taylor, PhD, Mübeccel Akdis, MD, PhD, Andrea Joss, PhD, Tunç Akkoç, PhD, Renate Wenig, PhD, Marco Colonna, MD, Isabelle Daigle, PhD, Egbert Flory, PhD, Kurt Blaser, PhD, Cezmi A. Akdis, MD Journal of Allergy and Clinical Immunology Volume 120, Issue 1, Pages 76-83 (July 2007) DOI: 10.1016/j.jaci.2007.04.004 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 IL-10 suppresses T-cell proliferation induced by CD28 and ICOS costimulations. A, Proliferation of CD45R0+ human T cells. ∗P < .05. B, Proliferation of CD4+ human T cells. C, PBMC proliferation. PPD, Purified protein derivative of Mycobacterium bovis; TT, tetanus toxoid. CFSE labeling of (D) PBMCs, (E) PBMCs in the presence of IL-2, and (F) natural killer cell and monocyte depleted PBMCs. The numbers indicate the percentage of cells at a certain number of divisions at day 5. u.s., Unstimulated control cells; Ag, antigen. One representative of 3 experiments is shown. Journal of Allergy and Clinical Immunology 2007 120, 76-83DOI: (10.1016/j.jaci.2007.04.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 IL-10 inhibits tyrosine phosphorylation of CD28 and ICOS, and the association of these costimulatory molecules with PI3-K via SHP-1. A, Expression of ICOS in human CD45RO+ T cells (top), tyrosine phosphorylation of ICOS (Ptyr, middle), and PI3-K association (bottom). B, Coimmunoprecipitation of SHP-1 with anti-Tyk2 mAb in human T cells. C, Expression of SHP-1 and its tyrosine phosphorylation in the presence of IL-10. A-C, The upper panels represent loading controls for the lower panels. D, IL-10–induced tyrosine phosphorylation and binding of SHP-1 to CD28 and ICOS. IP, Immunoprecipitation; IgH, Ig heavy chain. Data shown A-D represent 1 of at least 3 independent experiments. Journal of Allergy and Clinical Immunology 2007 120, 76-83DOI: (10.1016/j.jaci.2007.04.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Blocking of SHP-1 expression by siRNA nuclear transfection attenuated the suppressive effect of IL-10 on CD28 and ICOS. A, SHP-1 expression after transfection of human T cells with SHP-1 siRNA. B, Proliferation of transfected human T cells in the presence or absence of IL-10. ERK, Extracellular signal-related kinase; u.s., resting human T cells. One of 5 independent experiments with similar results is shown. ∗P < .05. Journal of Allergy and Clinical Immunology 2007 120, 76-83DOI: (10.1016/j.jaci.2007.04.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 IL-10 does not suppress CD28 and ICOS costimulations in SHP-1 DN overexpressing T cells. A, SHP-1 expression in SHP-1 DN transduced human CD4+ T cells. B, NGFR and GFP expression in transduced T cells. Solid curve, Control mAb. Open curve, GFP or NGFR expression. C, Proliferation of SHP-1 DN expressing human T cells. u.s., Resting human T cells. Results represent 1 of 3 independent experiments. ∗P < .05. Journal of Allergy and Clinical Immunology 2007 120, 76-83DOI: (10.1016/j.jaci.2007.04.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 SHP-1–deficient mouse T cells show increased proliferation with costimulation alone and do not respond to IL-10–mediated suppression. Proliferation of splenic CD4+ T cells from wild-type, IL-10–deficient (A) and SHP-1–deficient (me/me) (B) mice. B7RP-1, ICOS ligand. Data are representative of 2 experiments for each of A, 5 mice, and B, 3 mice. wt, Wild-type. Journal of Allergy and Clinical Immunology 2007 120, 76-83DOI: (10.1016/j.jaci.2007.04.004) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions