Imaging the Single Cell Dynamics of Cd4 Ϩ T Cell Activation by Dendritic Cells in Lymph Nodes

The adaptive immune response is initiated in secondary lymphoid organs by contact between antigen-bearing dendritic cells (DCs) and antigen-specific CD4 ϩ T cells. However, there is scant information regarding the single cell dynamics of this process in vivo. Using two-photon microscopy, we imaged the real-time behavior of naive CD4 ϩ T cells and in vivo–labeled DCs in lymph nodes during a robust T cell response. In the first 2 h after entry into lymph nodes, T cells made short-lived contacts with antigen-bearing DCs, each contact lasting an average of 11–12 min and occurring mainly on dendrites. Altered patterns of T cell motility during this early stage of antigen recognition promoted serial engagement with several adjacent DCs. Subsequently , T cell behavior progressed through additional distinct stages, including long-lived clusters, dynamic swarms, and finally autonomous migration punctuated by cell division. These observations suggest that the immunological synapse in native tissues is remarkably fluid, and that stable synapses form only at specific stages of antigen presentation to T cells. Furthermore, the serial nature of these interactions implies that T cells activate by way of multiple antigen recognition events.

[1]  M. Krummel,et al.  New views of the immunological synapse: variations in assembly and function. , 2004, Current opinion in immunology.

[2]  Mark M Davis,et al.  T cell killing does not require the formation of a stable mature immunological synapse , 2004, Nature Immunology.

[3]  Ronald N Germain,et al.  In vivo antigen presentation. , 2004, Current opinion in immunology.

[4]  Mark J. Miller,et al.  T cell repertoire scanning is promoted by dynamic dendritic cell behavior and random T cell motility in the lymph node. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Henrickson,et al.  T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases , 2004, Nature.

[6]  Mark M. Davis,et al.  T-cell-antigen recognition and the immunological synapse , 2003, Nature Reviews Immunology.

[7]  S. F. Actory,et al.  A stochastic view of lymphocyte motility and trafficking within the lymph node , 2003, Immunological reviews.

[8]  S. Granjeaud,et al.  The Strategy of T Cell Antigen-presenting Cell Encounter in Antigen-draining Lymph Nodes Revealed by Imaging of Initial T Cell Activation , 2003, The Journal of experimental medicine.

[9]  Ian Parker,et al.  Real-time imaging of lymphocytes in vivo. , 2003, Current opinion in immunology.

[10]  M. Jenkins,et al.  Antigen presentation to naive CD4 T cells in the lymph node , 2003, Nature Immunology.

[11]  C. Weaver,et al.  Restricted Clonal Expression of IL-2 By Naive T Cells Reflects Differential Dynamic Interactions with Dendritic Cells , 2003, The Journal of experimental medicine.

[12]  A. Rudensky,et al.  Distinct dendritic cell populations sequentially present antigen to CD4 T cells and stimulate different aspects of cell-mediated immunity. , 2003, Immunity.

[13]  J. Neilson,et al.  Calcium signalling in lymphocytes. , 2003, Current opinion in immunology.

[14]  Philippe Bousso,et al.  Dynamics of CD8+ T cell priming by dendritic cells in intact lymph nodes , 2003, Nature Immunology.

[15]  Mark J. Miller,et al.  Autonomous T cell trafficking examined in vivo with intravital two-photon microscopy , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[16]  B. Kyewski,et al.  Dynamic Changes During the Immune Response in T Cell–Antigen-presenting Cell Clusters Isolated from Lymph Nodes , 2003, The Journal of experimental medicine.

[17]  F. Sallusto,et al.  Opinion-decision making in the immune system: Progressive differentiation and selection of the fittest in the immune response , 2002, Nature Reviews Immunology.

[18]  Michael D. Cahalan,et al.  Two-photon tissue imaging: seeing the immune system in a fresh light , 2002, Nature Reviews Immunology.

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

[20]  Michael Loran Dustin,et al.  T-cell activation: a multidimensional signaling network. , 2002, Current opinion in cell biology.

[21]  D. McGavern,et al.  Molecular anatomy of antigen-specific CD8+ T cell engagement and synapse formation in vivo , 2002, Nature Immunology.

[22]  E. Bröcker,et al.  TCR triggering on the move: diversity of T‐cell interactions with antigen‐presenting cells , 2002, Immunological reviews.

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

[24]  Michael Loran Dustin,et al.  Immature CD4(+)CD8(+) thymocytes form a multifocal immunological synapse with sustained tyrosine phosphorylation. , 2002, Immunity.

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

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

[27]  Mark M. Davis,et al.  Dynamics of the immunological synapse: finding, establishing and solidifying a connection. , 2002, Current opinion in immunology.

[28]  R. Steinman,et al.  Dendritic cells: specialized and regulated antigen processing machines. , 2001, Cell.

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

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

[31]  E. Ingulli,et al.  Cutting Edge: In Vivo Identification of TCR Redistribution and Polarized IL-2 Production by Naive CD4 T Cells1 , 2001, The Journal of Immunology.

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

[33]  Michael L. Dustin,et al.  The immunological synapse and the actin cytoskeleton: molecular hardware for T cell signaling , 2000, Nature Immunology.

[34]  F. Sallusto,et al.  Two subsets of memory T lymphocytes with distinct homing potentials and effector functions , 1999, Nature.

[35]  S. Bromley,et al.  The immunological synapse: a molecular machine controlling T cell activation. , 1999, Science.

[36]  A J Young,et al.  The physiology of lymphocyte migration through the single lymph node in vivo. , 1999, Seminars in immunology.

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

[38]  A. Khoruts,et al.  In Vivo Detection of Dendritic Cell Antigen Presentation to CD4+ T Cells , 1997, The Journal of experimental medicine.

[39]  C. Hsieh,et al.  T cell genetic background determines default T helper phenotype development in vitro , 1995, The Journal of experimental medicine.

[40]  A. Khoruts,et al.  In vivo activation of antigen-specific CD4 T cells. , 2001, Annual review of immunology.