Random migration precedes stable target cell interactions of tumor-infiltrating T cells

The tumor microenvironment is composed of an intricate mixture of tumor and host-derived cells that engage in a continuous interplay. T cells are particularly important in this context as they may recognize tumor-associated antigens and induce tumor regression. However, the precise identity of cells targeted by tumor-infiltrating T lymphocytes (TILs) as well as the kinetics and anatomy of TIL-target cell interactions within tumors are incompletely understood. Furthermore, the spatiotemporal conditions of TIL locomotion through the tumor stroma, as a prerequisite for establishing contact with target cells, have not been analyzed. These shortcomings limit the rational design of immunotherapeutic strategies that aim to overcome tumor-immune evasion. We have used two-photon microscopy to determine, in a dynamic manner, the requirements leading to tumor regression by TILs. Key observations were that TILs migrated randomly throughout the tumor microenvironment and that, in the absence of cognate antigen, they were incapable of sustaining active migration. Furthermore, TILs in regressing tumors formed long-lasting (≥30 min), cognate antigen–dependent contacts with tumor cells. Finally, TILs physically interacted with macrophages, suggesting tumor antigen cross-presentation by these cells. Our results demonstrate that recognition of cognate antigen within tumors is a critical determinant of optimal TIL migration and target cell interactions, and argue against TIL guidance by long-range chemokine gradients.

[1]  D. Pardoll,et al.  Does the immune system see tumors as foreign or self? , 2003, Annual review of immunology.

[2]  T. Rothstein,et al.  Cytotoxic T lymphocyte sequential killing of immobilized allogeneic tumor target cells measured by time-lapse microcinematography. , 1978, Journal of immunology.

[3]  Mark M Davis,et al.  T cells use two directionally distinct pathways for cytokine secretion , 2006, Nature Immunology.

[4]  M. Kinouchi,et al.  Selective infiltration of CCR5+CXCR3+ T lymphocytes in human colorectal carcinoma , 2005, International journal of cancer.

[5]  Peter Friedl,et al.  Amoeboid leukocyte crawling through extracellular matrix: lessons from the Dictyostelium paradigm of cell movement , 2001, Journal of leukocyte biology.

[6]  Hao Shen,et al.  Control of Effector CD8+ T Cell Function by the Transcription Factor Eomesodermin , 2003, Science.

[7]  M. Mildner,et al.  Retinoic Acid Increases the Expression of p53 and Proapoptotic Caspases and Sensitizes Keratinocytes to Apoptosis , 2004, Cancer Research.

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

[9]  Ian Parker,et al.  In situ characterization of CD4+ T cell behavior in mucosal and systemic lymphoid tissues during the induction of oral priming and tolerance , 2005, The Journal of experimental medicine.

[10]  A. Chakraborty,et al.  Correction: Directed Migration of Positively Selected Thymocytes Visualized in Real Time , 2005, PLoS Biology.

[11]  D. Littman,et al.  A lineage-specific transcriptional silencer regulates CD4 gene expression during T lymphocyte development , 1994, Cell.

[12]  T. Blankenstein.  The role of tumor stroma in the interaction between tumor and immune system. , 2005, Current opinion in immunology.

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

[14]  Tobias Bonhoeffer,et al.  Live imaging of effector cell trafficking and autoantigen recognition within the unfolding autoimmune encephalomyelitis lesion , 2005, The Journal of experimental medicine.

[15]  Philippe Bousso,et al.  Dynamic behavior of T cells and thymocytes in lymphoid organs as revealed by two-photon microscopy. , 2004, Immunity.

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

[17]  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.

[18]  Michel Sadelain,et al.  Targeting tumours with genetically enhanced T lymphocytes , 2003, Nature Reviews Cancer.

[19]  E. Furth,et al.  CXC Chemokine Ligand 12 (Stromal Cell-Derived Factor 1α) and CXCR4-Dependent Migration of CTLs toward Melanoma Cells in Organotypic Culture1 , 2005, The Journal of Immunology.

[20]  Mark J. Miller,et al.  Antigen-Engaged B Cells Undergo Chemotaxis toward the T Zone and Form Motile Conjugates with Helper T Cells , 2005, PLoS biology.

[21]  Ronald N Germain,et al.  An extended vision for dynamic high-resolution intravital immune imaging. , 2005, Seminars in immunology.

[22]  A. Matter.  Microcinematographic and electron microscopic analysis of target cell lysis induced by cytotoxic T lymphocytes. , 1979, Immunology.

[23]  Christopher H Contag,et al.  Understanding immune cell trafficking patterns via in vivo bioluminescence imaging , 2002, Journal of cellular biochemistry. Supplement.

[24]  R. Schreiber,et al.  The three Es of cancer immunoediting. , 2004, Annual review of immunology.

[25]  J. Blattman,et al.  Cancer Immunotherapy: A Treatment for the Masses , 2004, Science.

[26]  U. V. von Andrian.  Intravital microscopy of the peripheral lymph node microcirculation in mice. , 1996, Microcirculation.

[27]  Alan Aderem,et al.  Dynamic Interactions of Macrophages with T Cells during Antigen Presentation , 1999, The Journal of experimental medicine.

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

[29]  K. D. Zylan,et al.  Article , 1996, Physiology & Behavior.

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

[31]  D. Jäger,et al.  Antigen‐specific immunotherapy and cancer vaccines , 2003, International journal of cancer.

[32]  Pau Serra,et al.  Visualizing regulatory T cell control of autoimmune responses in nonobese diabetic mice , 2006, Nature Immunology.

[33]  Noam Brown,et al.  The role of tumour‐associated macrophages in tumour progression: implications for new anticancer therapies , 2002, The Journal of pathology.

[34]  Eli Gilboa,et al.  The promise of cancer vaccines , 2004, Nature Reviews Cancer.

[35]  S. Hedrick,et al.  Elf-1 binds to a critical element in a second CD4 enhancer , 1994, Molecular and cellular biology.

[36]  R. Alon,et al.  Immune cell migration in inflammation: present and future therapeutic targets , 2005, Nature Immunology.

[37]  Sebastian Amigorena,et al.  Distinct T cell dynamics in lymph nodes during the induction of tolerance and immunity , 2004, Nature Immunology.

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

[39]  H. Schreiber,et al.  Bystander elimination of antigen loss variants in established tumors , 2004, Nature Medicine.

[40]  C. Sumen,et al.  ReviewIntravital Microscopy : Visualizing Immunity in Context , 2004 .

[41]  J. Segall,et al.  Intravital imaging of cell movement in tumours , 2003, Nature Reviews Cancer.

[42]  Michel C Nussenzweig,et al.  Stable T cell–dendritic cell interactions precede the development of both tolerance and immunity in vivo , 2005, Nature Immunology.

[43]  Michael Loran Dustin,et al.  The immunological synapse , 2002, Arthritis research.

[44]  J. Blattman,et al.  Adoptive immunotherapy: engineering T cell responses as biologic weapons for tumor mass destruction. , 2003, Cancer cell.

[45]  Franco Patrone,et al.  The use of dendritic cells in cancer immunotherapy. , 2008, Critical reviews in oncology/hematology.

[46]  S. Albelda,et al.  Cycloxygenase-2 Inhibition Augments the Efficacy of a Cancer Vaccine , 2006, Clinical Cancer Research.

[47]  Federica Sallusto,et al.  Chemoattractants and their receptors in homeostasis and inflammation. , 2004, Current opinion in immunology.

[48]  Steven A. Rosenberg,et al.  Adoptive-cell-transfer therapy for the treatment of patients with cancer , 2003, Nature Reviews Cancer.

[49]  Wolfgang Weninger,et al.  Selective imprinting of gut-homing T cells by Peyer's patch dendritic cells , 2003, Nature.

[50]  C. Sanderson.  The mechanism of T cell mediated cytotoxicity II. morphological studies of cell death by time-lapse microcinematography , 1976, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[51]  Z. Xiang,et al.  A method that allows easy characterization of tumor-infiltrating lymphocytes. , 2001, Journal of immunological methods.

[52]  R. Weissleder,et al.  In vivo high resolution three-dimensional imaging of antigen-specific cytotoxic T-lymphocyte trafficking to tumors. , 2003, Cancer research.