Project

Back to overview

Investigating the molecular factors controlling lymphocyte motility and activation by in vivo imaging

Applicant Stein Jens Volker
Number 135649
Funding scheme Project funding (Div. I-III)
Research institution Theodor Kocher Institut Medizinische Fakultät Universität Bern
Institution of higher education University of Berne - BE
Main discipline Immunology, Immunopathology
Start/End 01.04.2011 - 31.03.2014
Approved amount 478'184.00
Show all

All Disciplines (2)

Discipline
Immunology, Immunopathology
Cellular Biology, Cytology

Keywords (6)

Lymphocyte migration; Homeostatic chemokines; Twophoton microscopy; Chemokine receptor signaling ; T cell - DC interactions; Antiviral immune response

Lay Summary (English)

Lead
Lay summary

Lymphocyte trafficking and activation

 

. Within these organs, lymphocytes screen antigen-presenting cells for their cognate antigen. Recirculation is completed by migration of lymphocytes back to blood via lymphatic vessels. Continuous trafficking makes lymphocytes one of the most motile mammalian cell types, migrating hundreds of micrometer every day within tissue.spleen and lymph nodes through the body in search of foreign antigen. Lymphocytes are in permanent movement from blood into lymphoid organs such as continuous lymphocyte traffickingAn essential feature of the adaptive immune system is the

 

Chemokines are small secreted polypeptides highly expressed in spleen and lymph nodes and play an important role in the selective recruitment of blood-borne lymphocytes into lymphoid tissue. Also, chemokines are responsible for lymphocyte segregation into T and B cell areas inside lymphoid tissue. As an example, the chemokine CCL21 is highly expressed in the T cell area of lymph nodes, as well as on high endothelial venules (HEV), which serve as port of entry for circulating lymphocytes. CCL21 is a strong chemoattractant for naïve T cells and serves therefore as a central organizing molecule for lymphoid organs.chemokine receptors. Lymphocyte trafficking is a finely tuned mechanism, which is in large part regulated by adhesion receptors of the integrin family and G-protein-coupled receptor (GPCR), notably

 

Our research group works on three complementary lines of investigation:

 

, with a special emphasis on intracellular chemokine receptor signaling (small GTPases and their activators, etc) and the roles of adhesion molecules and stromal cells in this process;molecular mechanisms of lymphocyte migration to and within lymph nodes1. A major focus is the study of

 

during immune responses using transgenic lymphocytes lacking of one or more signaling or adhesion molecules; T cell-dendritic cell (DC) interactions2. We are examining

 

. With OPT, we are performing mesoscopic imaging of the internal 3D structure of lymph nodes during homeostasis and inflammation.optical projection tomography (OPT) and intravital microscopy, (IVM), multiplex twophoton microscopy (2PM) for a quantitative analysis of adaptive immune responses, mainly imaging techniques3. We are applying novel

.in vivoIn summary, we are combining cutting-edge microscopy and complementary techniques with transgenic models to examine the migratory and activation behavior of lymphocytes

Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
OPTiSPIM: integrating optical projection tomography in light sheet microscopy extends specimen characterization to nonfluorescent contrasts.
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.
Endothelial cell-specific lymphotoxin-β receptor signaling is critical for lymph node and high endothelial venule formation.
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.
Janus kinases 1 and 2 regulate chemokine-mediated integrin activation and naïve T-cell homing.
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, 1.
Maturation of Lymph Node Fibroblastic Reticular Cells from Myofibroblastic Precursors Is Critical for Antiviral Immunity.
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, 1.
Naive B cell trafficking is shaped by local chemokine availability and LFA-1-independent stromal interactions.
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, 1.
A global "imaging'' view on systems approaches in immunology.
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.
DOCK8 is a Cdc42 activator critical for interstitial dendritic cell migration during immune responses.
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.
HIV-1 Nef interferes with T-lymphocyte circulation through confined environments in vivo.
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.
Optical projection tomography reveals dynamics of HEV growth after immunization with protein plus CFA and features shared with HEVs in acute autoinflammatory lymphadenopathy.
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.
Quantitative measurements in 3-dimensional datasets of mouse lymph nodes resolve organ-wide functional dependencies.
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.
In vivo analysis of uropod function during physiological T cell trafficking.
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.

Collaboration

Group / person Country
Types of collaboration
ISTA, Vienna Austria (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Kyushu University Japan (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Centre for Genome Regulation Spain (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Exchange of personnel
University of Geneva Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Universität Lausanne Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
University of Heidelberg Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Kantonsspital St Gallen Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Research Infrastructure
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
WIRM meeting Talk given at a conference Intravital microscopy of immune responses 19.03.2014 Davos, Switzerland Ozga Aleksandra; Pieczyk Markus;
Symposium Talk given at a conference Intravital microscopy of immune responses 16.09.2013 Rotterdam, Netherlands Stein Jens Volker;
Seminar Individual talk Intravital microscopy of immune responses 23.04.2013 Basel, Switzerland Stein Jens Volker;
JST Crest meeting Talk given at a conference Intravital microscopy of immune responses 12.02.2013 Tokyo, Japan, Japan Stein Jens Volker;
Seminar Kyushu University Talk given at a conference Intravital microscopy of immune responses 05.02.2013 Fukuoka, Japan, Japan Stein Jens Volker;
BIC seminar Talk given at a conference Intravital microscopy of immune responses 28.11.2012 Bern, Switzerland, Switzerland Stein Jens Volker;
Seminar University of Heidelberg Talk given at a conference Intravital microscopy of immune responses 05.11.2012 Heidelberg, Germany, Germany Stein Jens Volker;
Seminar Instituto Humanitas Talk given at a conference Intravital microscopy of immune responses 05.10.2012 Milano, Italy, Italy Stein Jens Volker;
Seminar CNB Talk given at a conference Intravital microscopy of immune responses 12.06.2012 Madrid, Spain, Spain Stein Jens Volker;
AAI meeting Talk given at a conference Intravital microscopy of immune responses 04.05.2012 Boston, United States of America Stein Jens Volker;
World Immune regulation meeting Talk given at a conference Intravital microscopy of immune responses 19.03.2012 Davos, Switzerland, Switzerland Stein Jens Volker;
Laser sheet microscopy meeting Talk given at a conference Laser sheet microscopy 14.10.2011 Toulouse, France, France Stein Jens Volker;
Seminar University of Magdeburg Talk given at a conference Intravital microscopy of immune responses 28.04.2011 Magdeburg, Germany, Germany Stein Jens Volker;


Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions GenTage, Schaffhausen German-speaking Switzerland 2012

Awards

Title Year
Bangerter Rhyner foundation 2013
Novartis Foundation grant 2011

Associated projects

Number Title Start Funding scheme
120640 In vivo analysis of intracellular signaling pathways during lymphocyte trafficking and activation 01.04.2008 Project funding (Div. I-III)
153457 Visualizing molecular mechanisms of organ-specific T cell activation and immune surveillance 01.04.2014 Project funding (Div. I-III)
139232 3D Ultramicroscopy for biological imaging 01.12.2011 R'EQUIP
125447 Examining the function of lymphoid organ structure during antiviral immune responses using microscopic and mesoscopic imaging 01.06.2009 Sinergia
141918 Imaging-based Systems Biology Analysis of Lymph Node Structure and Function in Viral Infection 01.08.2012 Sinergia

Abstract

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.
-