Twophoton microscopy; Chemokine receptor signaling ; Lymphocyte migration; Antiviral immune response; Immune surveillance; T cell - DC interactions; Organ-specific immune surveillance
Stein Jens V, F Gonzalez Santiago (2017), Dynamic intravital imaging of cell-cell interactions in the lymph node., in The Journal of allergy and clinical immunology
, 139(1), 12-20.
Vladymyrov Mykhailo, Abe Jun, Moalli Federica, Stein Jens V, Ariga Akitaka (2016), Real-time tissue offset correction system for intravital multiphoton microscopy., in Journal of immunological methods
, 438, 35-41.
Ozga Aleksandra J, Moalli Federica, Abe Jun, Swoger Jim, Sharpe James, Zehn Dietmar, Kreutzfeldt Mario, Merkler Doron, Ripoll Jorge, Stein Jens V (2016), pMHC affinity controls duration of CD8+ T cell-DC interactions and imprints timing of effector differentiation versus expansion., in The Journal of experimental medicine
, 213(12), 2811-2829.
Köchl Robert, Thelen Flavian, Vanes Lesley, Brazão Tiago F, Fountain Kathryn, Xie Jian, Huang Chou-Long, Lyck Ruth, Stein Jens V, Tybulewicz Victor L J (2016), WNK1 kinase balances T cell adhesion versus migration in vivo., in Nature immunology
, 17(9), 1075-83.
Zanotti L, Angioni R, Calì B, Soldani C, Ploia C, Moalli F, Gargesha M, D'Amico G, Elliman S, Tedeschi G, Maffioli E, Negri A, Zacchigna S, Sarukhan A, Stein J V, Viola A (2016), Mouse mesenchymal stem cells inhibit high endothelial cell activation and lymphocyte homing to lymph nodes by releasing TIMP-1., in Leukemia
, 30(5), 1143-54.
Ackerknecht Markus, Hauser Mark A, Legler Daniel F, Stein Jens V (2015), In vivo TCR Signaling in CD4(+) T Cells Imprints a Cell-Intrinsic, Transient Low-Motility Pattern Independent of Chemokine Receptor Expression Levels, or Microtubular Network, Integrin, and Protein Kinase C Activity., in Frontiers in immunology
, 6, 297-297.
Moalli Federica, Proulx Steven T, Schwendener Reto, Detmar Michael, Schlapbach Christoph, Stein Jens V (2015), Intravital and whole-organ imaging reveals capture of melanoma-derived antigen by lymph node subcapsular macrophages leading to widespread deposition on follicular dendritic cells., in Frontiers in immunology
, 6, 114-114.
Stein Jens V (2015), T Cell Motility as Modulator of Interactions with Dendritic Cells., in Frontiers in immunology
, 6, 559-559.
Tang Fengyuan, Gill Jason, Ficht Xenia, Barthlott Thomas, Cornils Hauke, Schmitz-Rohmer Debora, Hynx Debby, Zhou Dawang, Zhang Lei, Xue Gongda, Grzmil Michal, Yang Zhongzhou, Hergovich Alexander, Hollaender Georg A, Stein Jens V, Hemmings Brian A, Matthias Patrick (2015), The kinases NDR1/2 act downstream of the Hippo homolog MST1 to mediate both egress of thymocytes from the thymus and lymphocyte motility., in Science signaling
, 8(397), 100-100.
Moalli Federica, Cupovic Jovana, Thelen Flavian, Halbherr Pascal, Fukui Yoshinori, Narumiya Shuh, Ludewig Burkhard, Stein Jens V (2014), Thromboxane A2 acts as tonic immunoregulator by preferential disruption of low-avidity CD4+ T cell-dendritic cell interactions., in The Journal of experimental medicine
, 211(13), 2507-17.
Naïve T cells are programmed to home to secondary lymphoid organs (SLOs), including spleen and peripheral lymph nodes (PLNs). There, they scan antigen-presenting cells (APCs), in particular dendritic cells (DCs), for the presence of cognate peptide-MHC (pMHC) complexes. Studies using intravital twophoton microscopy (2PM) of lymphoid tissue in live, anesthetized mice have revealed that interactions are carefully balanced to avoid activation of self-reactive T cells while triggering productive T cell - DC interactions to foreign pMHC. Once naïve T cells become activated, these cells divide and differentiate into effector cells. Some effector cells migrate to the site of inflammation, where a subset of effector memory T cells remains in tissue long after pathogen clearance. These tissue resident memory T cells (TRM) act as first line of defense upon pathogen re-exposure. On the other hand, follicular helper T cells (TFH) remain in lymphoid organs to provide B cell help in germinal centers and ensure the generation of high-affinity mAbs for protective immunity. In sum, a defining feature of T cells is their capacity to migrate through blood and lymph to lymphoid and non-lymphoid organs, where they engage in dynamic interactions with other hematopoietic and stromal cells. Here, we propose to use mouse models for the functional visualization of interactions of T cells, APCs and stromal cells in lymphoid and non-lymphoid organs during acute and memory responses, in sterile and infectious inflammation. We will carry out four complementary projects. First, we will perform an in-depth 2PM analysis of lymphoid stroma - T cell interactions for efficient motility. Second, we will use functional 2PM imaging of signaling reporters to visualize checkpoints of TFH formation and compare these with a mutant strain with impaired TFH formation, using gene expression analysis and 2PM. We will also examine the effect of HIV-1 pathogenicity factor Nef for TFH function Third, we will investigate CD8+ T cell activation with cognate and altered peptide ligands and examine their impact on encounters with DCs arriving later during immune responses using flow cytometry and 2PM. The fourth and major part focuses on the role of intracellular and extracellular signaling molecules governing T cell immune surveillance through stromal cell and APC interactions in steady state, as well as during acute and memory phase of virus infections. Our model organs are PLNs for central memory T cells, as well as salivary gland and skin for effector memory T cells. Using transgenic mouse models and pharmacological inhibitors, which interfere with distinct key intracellular factors of the small GTPases family and integrins, we will make an in-depth 2PM and flow cytometry analysis of organ-specific migration requirements. An important part of this aim is to assign site-specific roles for signaling modules to efficient motility in tissues with increasing epithelial content (“tissue density”) and to correlate our findings with resolution of antimicrobial infection. In sum, our proposal builds on our established line of research of visualizing immune responses, while taking into account infectious models and the tissue context of immune surveillance.