Lymphocyte migration; Homeostatic chemokines; Twophoton microscopy; Chemokine receptor signaling ; T cell - DC interactions; Antiviral immune response
Mayer Jürgen, Robert-Moreno Alexandre, Danuser Renzo, Stein Jens V, Sharpe James, Swoger Jim (2014), OPTiSPIM: integrating optical projection tomography in light sheet microscopy extends specimen characterization to nonfluorescent contrasts., in Optics letters
, 39(4), 1053-6.
Onder Lucas, Danuser Renzo, Scandella Elke, Firner Sonja, Chai Qian, Hehlgans Thomas, Stein Jens V, Ludewig Burkhard (2013), Endothelial cell-specific lymphotoxin-β receptor signaling is critical for lymph node and high endothelial venule formation., in The Journal of experimental medicine
, 210(3), 465-73.
Pérez-Rivero Gema, Cascio Graciela, Soriano Silvia Fernández, Sanz Alvaro Gil, de Guinoa Julia Sáez, Rodríguez-Frade José Miguel, Gomariz Rosa P, Holgado Borja L, Cabañas Carlos, Carrasco Yolanda R, Stein Jens V, Mellado Mario (2013), Janus kinases 1 and 2 regulate chemokine-mediated integrin activation and naïve T-cell homing., in European journal of immunology
Chai Qian, Onder Lucas, Scandella Elke, Gil-Cruz Cristina, Perez-Shibayama Christian, Cupovic Jovana, Danuser Renzo, Sparwasser Tim, Luther Sanjiv A, Thiel Volker, Rülicke Thomas, Stein Jens V, Hehlgans Thomas, Ludewig Burkhard (2013), Maturation of Lymph Node Fibroblastic Reticular Cells from Myofibroblastic Precursors Is Critical for Antiviral Immunity., in Immunity
Coelho Fernanda M, Natale Daniela, Soriano Silvia F, Hons Miroslav, Swoger Jim, Mayer Jürgen, Danuser Renzo, Scandella Elke, Pieczyk Markus, Zerwes Hans-Günter, Junt Tobias, Sailer Andreas W, Ludewig Burkhard, Sharpe James, Figge Marc Thilo, Stein Jens V (2013), Naive B cell trafficking is shaped by local chemokine availability and LFA-1-independent stromal interactions., in Blood
Ludewig Burkhard, Stein Jens V, Sharpe James, Cervantes-Barragan Luisa, Thiel Volker, Bocharov Gennady (2012), A global "imaging'' view on systems approaches in immunology., in European journal of immunology
, 42(12), 3116-25.
Harada Yosuke, Tanaka Yoshihiko, Terasawa Masao, Pieczyk Markus, Habiro Katsuyoshi, Katakai Tomoya, Hanawa-Suetsugu Kyoko, Kukimoto-Niino Mutsuko, Nishizaki Tomoko, Shirouzu Mikako, Duan Xuefeng, Uruno Takehito, Nishikimi Akihiko, Sanematsu Fumiyuki, Yokoyama Shigeyuki, Stein Jens V, Kinashi Tatsuo, Fukui Yoshinori (2012), DOCK8 is a Cdc42 activator critical for interstitial dendritic cell migration during immune responses., in Blood
, 119(19), 4451-61.
Stolp Bettina, Imle Andrea, Coelho Fernanda Matos, Hons Miroslav, Gorina Roser, Lyck Ruth, Stein Jens V, Fackler Oliver T (2012), HIV-1 Nef interferes with T-lymphocyte circulation through confined environments in vivo., in Proceedings of the National Academy of Sciences of the United States of America
, 109(45), 18541-6.
Kumar Varsha, Chyou Susan, Stein Jens V, Lu Theresa T (2012), Optical projection tomography reveals dynamics of HEV growth after immunization with protein plus CFA and features shared with HEVs in acute autoinflammatory lymphadenopathy., in Frontiers in immunology
, 3, 282-282.
Mayer Jürgen, Swoger Jim, Ozga Aleksandra J, Stein Jens V, Sharpe James (2012), Quantitative measurements in 3-dimensional datasets of mouse lymph nodes resolve organ-wide functional dependencies., in Computational and mathematical methods in medicine
, 2012, 128431-128431.
Soriano Silvia F, Hons Miroslav, Schumann Kathrin, Kumar Varsha, Dennier Timo J, Lyck Ruth, Sixt Michael, Stein Jens V (2011), In vivo analysis of uropod function during physiological T cell trafficking., in Journal of immunology (Baltimore, Md. : 1950)
, 187(5), 2356-64.
Continuous trafficking to lymphoid tissue and cellular interactions with Antigen (Ag)-presenting cells (APCs) are hallmarks of lymphocyte biology. Guided by the homeostatic chemokines CCL21 and CXCL13, naïve T and B cells constitutively home to secondary lymphoid organs including spleen and peripheral lymph nodes (PLNs), where they systematically scan APCs, including dendritic cells (DCs), for presence of their cognate Ag. Recent technological advances through the combination of transgenic mouse lines and intravital twophoton microscopy (2PM), which enables the direct visualization of lymphocyte behavior deep within lymphoid tissue, has granted the immunological research community unprecedented insights into the molecular orchestration of the adaptive immune response. Here, we propose to continue our previous research on the molecular mechanisms of lymphocyte trafficking and activation by following three lines of research: first, we will use in vitro assays combined with intravital 2PM of mouse lymphoid tissue to investigate the function of intra- and extracellular motility-inducing factors during lymphocyte trafficking (Project 1). Specifically, we will examine the migratory behavior of lymphocytes genetically engineered to lack members of the DOCK family of guanine exchange factors, Tec kinases, phosphoinositide-3-kinases (PI3K) and the CXCL13-receptor CXCR5, in addition to pharmacological inhibition of signaling pathways downstream of chemokine receptors. The aim of this project is to obtain a comprehensive overview over key factors governing physiological lymphocyte trafficking. Second, we will examine the roles of DOCK and PI3K proteins, as well as LFA-1 - ICAM and thromboxane A2, during pMHC-driven dynamic interactions between Ag-specific CD4+ T cells and DCs (Project 2). We will systematically alter the pMHC concentration on DCs, which influences the T cell - DC contact duration, and investigate by intravital 2PM and flow cytometry the function of the intracellular factors DOCK2, DOCK8, PI3K? and PI3Kd during the formation of productive T cell - DC encounters and effector cell generation. We will also modify the adhesive strength of T cell - DC interaction by using hypo- and hyperadhesive LFA-1 - and ICAM-1/2 mutant leukocytes and correlate these data with the efficacy of effector T cell generation. Furthermore, we will investigate T cells with altered chemokinetic behavior in absence of the thromboxane A2 receptor, and its role for the orchestration of T cell - DC interactions. Third, we will use selective plane illumination microscopy (SPIM) to investigate the accumulation of virus-specific T cells during an immune response (Project 3). SPIM is a recently developed mesoscopic imaging technology, which we have adapted to allow detection of rare Ag-specific lymphocytes inside entire PLNs. Using SPIM, we will examine the dynamic accumulation of these cells during antiviral immune responses and correlate these findings with the efficiency of virus elimination.In summary, our research plan proposes to combine functional assays using genetically modified or pharmacologically inhibited lymphocytes with intravital and mesoscopic imaging to uncover the role for intra-and extracellular factors, which govern trafficking and interactions with APCs.