Continuous T cell receptor signaling required for synapse maintenance and full effector potential

Although signals through the T cell receptor (TCR) are essential for the initiation of T helper cell activation, it is unclear what function such signals have during the prolonged T cell–antigen-presenting cell contact. Here we simultaneously tracked TCR-CD3 complex and phosphoinositide 3-kinase activity in single T cells using three-dimensional video microscopy. Despite rapid internalization of most of the TCR-CD3, TCR-dependant signaling was still evident up to 10 h after conjugate formation. Blocking this interaction caused dissolution of the synapse and proportional reductions in interleukin 2 production and cellular proliferation. Thus TCR signaling persists for hours, has a cumulative effect and is necessary for the maintenance of the immunological synapse.

[1]  A. Lanzavecchia,et al.  The duration of antigenic stimulation determines the fate of naive and effector T cells. , 1998, Immunity.

[2]  G. Griffiths,et al.  The immunological synapse of CTL contains a secretory domain and membrane bridges. , 2001, Immunity.

[3]  Mark M. Davis,et al.  Direct observation of ligand recognition by T cells , 2002, Nature.

[4]  S. Ward,et al.  Phosphoinositide 3-kinases in T lymphocyte activation. , 2001, Current opinion in immunology.

[5]  D. Jeanfavre,et al.  Molecular regulation of the interaction between leukocyte function-associated antigen-1 and soluble ICAM-1 by divalent metal cations. , 1998, Journal of immunology.

[6]  Anjana Rao,et al.  Gene regulation mediated by calcium signals in T lymphocytes , 2001, Nature Immunology.

[7]  Mark M. Davis,et al.  Imaging synapse formation during thymocyte selection: inability of CD3zeta to form a stable central accumulation during negative selection. , 2002, Immunity.

[8]  Stephen P. Schoenberger,et al.  Naïve CTLs require a single brief period of antigenic stimulation for clonal expansion and differentiation , 2001, Nature Immunology.

[9]  T. Zal,et al.  Inhibition of T cell receptor-coreceptor interactions by antagonist ligands visualized by live FRET imaging of the T-hybridoma immunological synapse. , 2002, Immunity.

[10]  R. Steinman,et al.  Dendritic cells and the control of immunity , 1998, Nature.

[11]  D. Cantrell,et al.  Sustained and dynamic inositol lipid metabolism inside and outside the immunological synapse , 2002, Nature Immunology.

[12]  Stephen P. Schoenberger,et al.  T-cell help for cytotoxic T lymphocytes is mediated by CD40–CD40L interactions , 1998, Nature.

[13]  A. B. Lyons,et al.  Determination of lymphocyte division by flow cytometry. , 1994, Journal of immunological methods.

[14]  Chyung-Ru Wang,et al.  Helper T cell differentiation is controlled by the cell cycle. , 1998, Immunity.

[15]  R. Steinman,et al.  Antigen processing for amateurs and professionals. , 1998, Trends in cell biology.

[16]  Michael Loran Dustin,et al.  T Cell Receptor Signaling Precedes Immunological Synapse Formation , 2002, Science.

[17]  E. Bröcker,et al.  Antigen presentation in extracellular matrix: interactions of T cells with dendritic cells are dynamic, short lived, and sequential. , 2000, Immunity.

[18]  A. Weiss,et al.  Signal transduction by the TCR for antigen. , 2000, Current opinion in immunology.

[19]  J. Harty,et al.  CD8+ T cell effector mechanisms in resistance to infection. , 2000, Annual review of immunology.

[20]  L. Turka,et al.  The Proliferative Capacity of Individual Naive CD4+T Cells Is Amplified by Prolonged T Cell Antigen Receptor Triggering , 2002, The Journal of experimental medicine.

[21]  Sabine Stoll,et al.  Imaging of T‐cell interactions with antigen presenting cells in culture and in intact lymphoid tissue , 2002, Immunological reviews.

[22]  R. Basch,et al.  Early thymic regeneration after irradiation. , 1994, Developmental and comparative immunology.

[23]  Mark J. Miller,et al.  Two-Photon Imaging of Lymphocyte Motility and Antigen Response in Intact Lymph Node , 2002, Science.

[24]  Colin R. F. Monks,et al.  Three-dimensional segregation of supramolecular activation clusters in T cells , 1998, Nature.

[25]  S. Dzik,et al.  The immunological synapse: A molecular machine controlling T cell activation , 2000 .

[26]  K. Ozato,et al.  Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens. , 1980, Journal of immunology.

[27]  S. Bromley,et al.  Identification of self through two-dimensional chemistry and synapses. , 2001, Annual review of cell and developmental biology.

[28]  Mark M. Davis,et al.  Determination of the Relationship Between T Cell Responsiveness and the Number of MHC-Peptide Complexes Using Specific Monoclonal Antibodies1 , 2000, The Journal of Immunology.

[29]  S. Grinstein,et al.  Localized Biphasic Changes in Phosphatidylinositol-4,5-Bisphosphate at Sites of Phagocytosis , 2000, The Journal of cell biology.

[30]  Michael S. Kuhns,et al.  CTLA-4: new insights into its biological function and use in tumor immunotherapy , 2002, Nature Immunology.

[31]  P. Wernet,et al.  Dissection of human allostimulatory determinants with cloned T cells: stimulation inhibition by monoclonal antibodies TU22, 34, 35, 36, 37, 39, 43, and 58 against distinct human MHC class II molecules. , 1985, Human immunology.

[32]  Tetsuo Yamazaki,et al.  T cell receptor ligation induces the formation of dynamically regulated signaling assemblies , 2002, The Journal of cell biology.

[33]  J. Bluestone,et al.  Molecular basis of T cell inactivation by CTLA-4. , 1998, Science.

[34]  Richard A. Flavell,et al.  Help for cytotoxic-T-cell responses is mediated by CD40 signalling , 1998, Nature.

[35]  J. Egen,et al.  Cytotoxic T lymphocyte antigen-4 accumulation in the immunological synapse is regulated by TCR signal strength. , 2002, Immunity.

[36]  A. Lanzavecchia,et al.  Sustained signaling leading to T cell activation results from prolonged T cell receptor occupancy. Role of T cell actin cytoskeleton , 1995, The Journal of experimental medicine.

[37]  G. Bismuth,et al.  Imaging antigen-induced PI3K activation in T cells , 2002, Nature Immunology.

[38]  A. Lanzavecchia,et al.  Degradation of  T Cell Receptor (TCR)–CD3-ζ Complexes after Antigenic Stimulation , 1997, The Journal of experimental medicine.

[39]  M. Davis,et al.  Differential clustering of CD4 and CD3zeta during T cell recognition. , 2000, Science.

[40]  S. Grinstein,et al.  Restricted Accumulation of Phosphatidylinositol 3-Kinase Products in a Plasmalemmal Subdomain during Fcγ Receptor-Mediated Phagocytosis , 2001, The Journal of cell biology.

[41]  J. Miller,et al.  Cross-presentation: a general mechanism for CTL immunity and tolerance. , 1998, Immunology today.

[42]  Susan M. Kaech,et al.  Memory CD8+ T cell differentiation: initial antigen encounter triggers a developmental program in naïve cells , 2001, Nature Immunology.

[43]  V. Moy,et al.  Direct evidence for two affinity states for lymphocyte function-associated antigen 1 on activated T cells. , 1993, The Journal of biological chemistry.

[44]  J. Unkeless.  Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors , 1979, The Journal of experimental medicine.

[45]  D. Wiest,et al.  On the dynamics of TCR:CD3 complex cell surface expression and downmodulation. , 2000, Immunity.

[46]  A. Lanzavecchia,et al.  The interplay between the duration of TCR and cytokine signaling determines T cell polarization , 1999, European journal of immunology.

[47]  C. Monks,et al.  Small splenic B cells that bind to antigen-specific T helper (Th) cells and face the site of cytokine production in the Th cells selectively proliferate: immunofluorescence microscopic studies of Th-B antigen- presenting cell interactions , 1994, The Journal of experimental medicine.

[48]  R. Germain,et al.  Dynamic Imaging of T Cell-Dendritic Cell Interactions in Lymph Nodes , 2002, Science.

[49]  A. Lanzavecchia,et al.  Serial triggering of many T-cell receptors by a few peptide–MHC complexes , 1995, Nature.