Project

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Examining the function of lymphoid organ structure during antiviral immune responses using microscopic and mesoscopic imaging

Applicant Stein Jens Volker
Number 125447
Funding scheme Sinergia
Research institution Theodor Kocher Institut Medizinische Fakultät Universität Bern
Institution of higher education University of Berne - BE
Main discipline Immunology, Immunopathology
Start/End 01.06.2009 - 31.07.2012
Approved amount 1'500'000.00
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All Disciplines (2)

Discipline
Immunology, Immunopathology
Cellular Biology, Cytology

Keywords (4)

Viral infection; peripheral lymph nodes; microscopic and macroscopic imaging; immune system

Lay Summary (English)

Lead
Lay summary
The adaptive immune system protects us from infectious viruses and other microbes. Cells of the adaptive immune system - lymphocytes and antigen-presenting cells (APC) - are highly organized in strategic tissue locations throughout the body, the secondary lymphoid organs.
The roles of SLO are i) to filter pathogens from blood and lymph, thereby retaining their spread through the body; ii) to present pathogens or derived peptides to antigen-specific lymphocytes; and iii) to create microenvironments, which regulate the differentiation and survival of antigen-specific lymphocytes.
The stromal cells which form the scaffold of T and B cell zones, the fibroblastic reticular cells (FRC) and follicular dendritic cells (FDC), respectively, are now recognized to participate in all of these processes.
Novel imaging techniques such as twophoton microscopy (2PM), which allows to directly observe the dynamic behavior of lymphocytes in PLN of live, anesthetized mice, have uncovered a role of FRC as contact guidance cues for migrating lymphocytes. Nonetheless, there is a lack of scientific knowledge on the role of stromal cells in homeostasis and antiviral immunity. Here, we propose to develop new transgenic mouse models to study the role of T cell zone stromal cells during homeostasis and antiviral immune responses. These studies will include intense collaboration between the groups of Dr. Stein (Subproject A - 2PM and lymphocyte trafficking) and Prof. Ludewig (Subproject B - viral immunology and transgenic mouse development).
SLO are dynamic structures, which expand and decrease in size in the course of an immune response. As the mechanisms of stromal restructuring have remained elusive, a comprehensive and quantitative analysis of the global 3D SLO anatomy using mesoscopic imaging techniques, such as Optical Projection Tomography (OPT) and selective plane illumination microscopy (SPIM) is highly desirable.We therefore propose to develop a hybrid OPT/SPIM scanner in collaboration with Dr. Sharpe, who had previously developed OPT. This project will entail a close collaboration with Dr. Stein (Subproject A) and Dr. Sharpe (Subproject C), with an emphasis on the global visualization of stromal cells in homeostasis and antiviral immune responses in transgenic mouse models developed by Prof. Ludewig (Subproject B). Using the newly developed OPT/SPIM scanner, these studies will also include a comprehensive analysis of morphogenetic factors governing vascular remodeling in antiviral immunity. In summary, our proposal aims to address the immunological relevance of lympoid stromal elements with novel imaging methods.

Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Image formation by linear and nonlinear digital scanned light-sheet fluorescence microscopy with Gaussian and Bessel beam profiles.
Olarte Omar E, Licea-Rodriguez Jacob, Palero Jonathan A, Gualda Emilio J, Artigas David, Mayer Jürgen, Swoger Jim, Sharpe James, Rocha-Mendoza Israel, Rangel-Rojo Raul, Loza-Alvarez Pablo (2012), Image formation by linear and nonlinear digital scanned light-sheet fluorescence microscopy with Gaussian and Bessel beam profiles., in Biomedical optics express, 3(7), 1492-505.
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.
Simulation of the interferon-mediated protective field in lymphoid organs with their spatial and functional organization taken into consideration.
Bocharov G A, Danilov A A, Vassilevski Yu V, Marchuk G I, Chereshnev V A, Ludewig B (2011), Simulation of the interferon-mediated protective field in lymphoid organs with their spatial and functional organization taken into consideration., in Doklady biological sciences : proceedings of the Academy of Sciences of the USSR, Biological science, 439, 194-6.
Two ways to use imaging: focusing directly on mechanism, or indirectly via behaviour?
Sharpe James (2011), Two ways to use imaging: focusing directly on mechanism, or indirectly via behaviour?, in Current opinion in genetics & development, 21(5), 523-9.
A systems immunology approach to plasmacytoid dendritic cell function in cytopathic virus infections.
Bocharov Gennady, Züst Roland, Cervantes-Barragan Luisa, Luzyanina Tatyana, Chiglintsev Egor, Chereshnev Valery A, Thiel Volker, Ludewig Burkhard (2010), A systems immunology approach to plasmacytoid dendritic cell function in cytopathic virus infections., in PLoS pathogens, 6(7), 1001017-1001017.
Comprehensive analysis of lymph node stroma-expressed Ig superfamily members reveals redundant and nonredundant roles for ICAM-1, ICAM-2, and VCAM-1 in lymphocyte homing.
Boscacci Rémy T, Pfeiffer Friederike, Gollmer Kathrin, Sevilla Ana Isabel Checa, Martin Ana Maria, Soriano Silvia Fernandez, Natale Daniela, Henrickson Sarah, von Andrian Ulrich H, Fukui Yoshinori, Mellado Mario, Deutsch Urban, Engelhardt Britta, Stein Jens V (2010), Comprehensive analysis of lymph node stroma-expressed Ig superfamily members reveals redundant and nonredundant roles for ICAM-1, ICAM-2, and VCAM-1 in lymphocyte homing., in Blood, 116(6), 915-25.
Critical roles for Rac GTPases in T-cell migration to and within lymph nodes.
Faroudi Mustapha, Hons Miroslav, Zachacz Agnieszka, Dumont Celine, Lyck Ruth, Stein Jens V, Tybulewicz Victor L J (2010), Critical roles for Rac GTPases in T-cell migration to and within lymph nodes., in Blood, 116(25), 5536-47.
Global lymphoid tissue remodeling during a viral infection is orchestrated by a B cell-lymphotoxin-dependent pathway.
Kumar Varsha, Scandella Elke, Danuser Renzo, Onder Lucas, Nitschké Maximilian, Fukui Yoshinori, Halin Cornelia, Ludewig Burkhard, Stein Jens V (2010), Global lymphoid tissue remodeling during a viral infection is orchestrated by a B cell-lymphotoxin-dependent pathway., in Blood, 115(23), 4725-33.
IL-7-producing stromal cells are critical for lymph node remodeling.
Onder Lucas, Narang Priyanka, Scandella Elke, Chai Qian, Iolyeva Maria, Hoorweg Kerim, Halin Cornelia, Richie Ellen, Kaye Paul, Westermann Jürgen, Cupedo Tom, Coles Mark, Ludewig Burkhard, IL-7-producing stromal cells are critical for lymph node remodeling., in Blood.

Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions Biologie heute German-speaking Switzerland 2011

Associated projects

Number Title Start Funding scheme
128786 Laser scanning confocal microscope for in vivo and in situ imaging 01.12.2009 R'EQUIP
137127 ProDoc Cell Migration Research Module I: Immune cell migration in immunosurveillance and inflammation 01.10.2011 ProDoc
177208 Defining the identity and differentiation pathways of the immune-stimulating fibroblastic tumor stroma 01.01.2018 Sinergia
139232 3D Ultramicroscopy for biological imaging 01.12.2011 R'EQUIP
120640 In vivo analysis of intracellular signaling pathways during lymphocyte trafficking and activation 01.04.2008 Project funding (Div. I-III)
135649 Investigating the molecular factors controlling lymphocyte motility and activation by in vivo imaging 01.04.2011 Project funding (Div. I-III)
141918 Imaging-based Systems Biology Analysis of Lymph Node Structure and Function in Viral Infection 01.08.2012 Sinergia

Abstract

The adaptive immune system protects us from infectious viruses and other microbes. Cells of the adaptive immune system - lymphocytes and antigen-presenting cells (APC) - are highly organized in strategic tissue locations throughout the body, the secondary lymphoid organs (SLO), such as peripheral lymph nodes (PLN) and spleen. The roles of SLO are i) to filter pathogens from blood and lymph, thereby retaining their spread through the body; ii) to present pathogens or derived peptides to antigen-specific lymphocytes; and iii) to create microenvironments, which regulate the differentiation and survival of antigen-specific lymphocytes. Inside SLO, T and B lymphocytes separate into particular microenvironments created by stromal cells, where they scan APC with high motility. When specific antigens are encountered, the trafficking patterns of lymphocytes change, accompanied by a stromal reorganization of the entire SLO structure. T and B cells start interacting for efficient immune responses, while affected SLO can increase several fold in size. This may ultimately result in a temporary loss of defined T and B cell zones and immune responsiveness.The stromal cells which form the scaffold of T and B cell zones, the fibroblastic reticular cells (FRC) and follicular dendritic cells (FDC), respectively, are now recognized to participate in all of these processes. Novel imaging techniques such as twophoton microscopy (2PM), which allows to directly observe the dynamic behavior of lymphocytes and APC in PLN of live, anesthetized mice, have uncovered a role of FRC as contact guidance cues for migrating lymphocytes. Furthermore, FRC secrete promigratory and prosurvival factors and may be involved in antigen presentation. Nonetheless, there are significant unmet needs in this research field, encompassing i) lack of scientific knowledge on the role of stromal cells in homeostasis and antiviral immunity, ii) lack of research tools which permit in vivo analysis of stromal cell biology, and iii) lack of researchers with expertise in the field to address these and future open questions. Here, we propose to develop new transgenic mouse models to study the role of T cell zone stromal cells during homeostasis and antiviral immune responses. These studies will include intense collaboration between the groups of Dr. Stein (Subproject A - 2PM and lymphocyte trafficking) and Prof. Ludewig (Subproject B - viral immunology and transgenic mouse development). By specifically depleting FRC, we will examine their role during lymphocyte migration and cellular interactions during homeostasis, antiviral immunity, and for T cell - APC interactions. Furthermore, the role of FRC for the decision-making between tolerance and immunity will be investigated.SLO are dynamic structures which expand and decrease in size in the course of an immune response. As the mechanisms of stromal restructuring have remained elusive, a comprehensive and quantitative analysis of the global 3D SLO anatomy using mesoscopic imaging techniques, such as Optical Projection Tomography (OPT) and selective plane illumination microscopy (SPIM) is highly desirable. In yet unpublished experiments, Dr. Stein’s group, in collaboration with Prof. Ludewig, adapted OPT to the analysis of entire PLN during homeostasis and antiviral immune responses. These OPT-generated data provide unprecedented insights into the dynamic regulation of vascular networks and B cell zones during adaptive immunity. However, intrinsic limitations of the commercial OPT scanner imposes restrictions on resolution and available detection channels, such that visualization of smaller structures, including the stromal scaffold, has could not be achieved yet. We therefore propose to overcome these restrictions through the development of a hybrid OPT/SPIM scanner in collaboration with Dr. Sharpe, who had previously developed OPT. This project will entail a close collaboration with Dr. Stein (Subproject A) and Dr. Sharpe (Subproject C), with an emphasis on the global visualization of stromal cells in homeostasis and antiviral immune responses in transgenic mouse models developed by Prof. Ludewig (Subproject B). Using the newly developed OPT/SPIM scanner, these studies will also include a comprehensive analysis of morphogenetic factors governing vascular remodeling in antiviral immunity. In summary, our proposal aims to address the immunological relevance of stromal elements of PLN with microscopic (2PM) and mesoscopic (OPT/SPIM) imaging. As SLO are the prime site for therapeutic and prophylactic immune interventions, elucidating the cellular and molecular mechanisms of SLO plasticity may allow to identify new avenues for therapeutic and preventive immunotherapy.
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