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Helmholtz Center for Infection Research

6 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Direct Observation of Phagocytosis and NET-formation by Neutrophils in Infected Lungs using 2-photon Microscopy

Mike Hasenberg¹, Anja Köhler¹, Susanne Bonifatius¹, Andreas Jeron², Matthias Gunzer¹

¹Institute for Molecular and Clinical Immunology, Otto-von-Guericke University Magdeburg, ²Department of Immunoregulation, Helmholtz Center for Infection Research

We show, how to use 2-photon microscopy for the observation of the dynamics of neutrophil granulocytes in infected lungs while they phagocytose pathogens or produce neutrophil extracellular traps (NETs).

Immunology and Infection

Flow Cytometric Isolation of Primary Murine Type II Alveolar Epithelial Cells for Functional and Molecular Studies

Marcus Gereke^1,2, Andrea Autengruber^1,2, Lothar Gröbe³, Andreas Jeron², Dunja Bruder^1,2, Sabine Stegemann-Koniszewski¹

¹Research Group Immune Regulation, Helmholtz Centre for Infection Research, ²Research Group Infection Immunology, Institute of Medical Microbiology, Otto-von-Guericke University , ³Department of Experimental Immunology, Helmholtz Centre for Infection Research

We describe the rapid isolation of primary murine type II alveolar epithelial cells (AECII) by flow cytometric negative selection. These AECII show high viability and purity and are suitable for a wide range of functional and molecular studies regarding their role in respiratory conditions such as autoimmune or infectious diseases.

JoVE Journal

Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange

Tim Pieters^1,2,3,4, Lieven Haenebalcke^1,2, Kenneth Bruneel^1,2,4, Niels Vandamme^1,2,4, Tino Hochepied^1,2, Jolanda van Hengel⁵, Dagmar Wirth⁶, Geert Berx^1,2,4, Jody J. Haigh⁷, Frans van Roy^1,2,4, Steven Goossens^1,2,3,4

¹Department of Biomedical Molecular Biology, Ghent University, ²Inflammation Research Center, VIB, ³Center for Medical Genetics, Ghent University Hospital, ⁴Cancer Research Institute Ghent (CRIG), ⁵Department of Basic Medical Sciences, Faculty of Medicine and Health Sciences, Ghent University, ⁶Helmholtz Center for Infection Research, ⁷Mammalian Functional Genetics Laboratory, Division of Blood Cancers, Australian Centre for Blood Diseases, Department of Clinical Haematology, Monash University and Alfred Health Alfred Centre

Proteins often contain multiple domains that can exert different cellular functions. Gene knock-outs (KO) do not consider this functional diversity. Here, we report a recombination-mediated cassette exchange (RMCE)-based structure-function approach in KO embryonic stem cells that allows for the molecular dissection of various functional domains or variants of a protein.

Neuroscience

Optimized Management of Endovascular Treatment for Acute Ischemic Stroke

Katharina Schregel^1,2, Daniel Behme¹, Ioannis Tsogkas¹, Michael Knauth¹, Ilko Maier³, André Karch⁴, Rafael Mikolajczyk^4,5, Mathias Bähr³, Jörn Schäper⁶, José Hinz⁶, Jan Liman³, Marios-Nikos Psychogios¹

¹Institute of Neuroradiology, University Medical Center Goettingen, ²Department of Radiology, Brigham and Women's Hospital and Harvard Medical School, ³Department of Neurology, University Medical Center Goettingen, ⁴Department of Epidemiology, Helmholtz Center for Infection Research, ⁵Institute of Medical Epidemiology, Biostatistics and Informatics, Martin-Luther-University Halle-Wittenberg, ⁶Department of Anesthesiology, University Medical Center Goettingen

The outcome of patients with acute ischemic stroke depends on swift restoration of cerebral blood flow. This protocol aims at optimizing the management of such patients by minimizing peri-procedural timings and rendering the time from hospital admission to reperfusion as short as possible.

Immunology and Infection

Pneumococcus Infection of Primary Human Endothelial Cells in Constant Flow

Hilger Jagau^1,2, Ina-Kristin Behrens^1,3, Michael Steinert^1,4, Simone Bergmann¹

¹Institut für Mikrobiologie, Technische Universität Braunschweig, ²Devision of Infection Medicine, Department of Clinical Science, Lund University, ³Medical Microbiology and Hospital Epidemiology, Max von Pettenkofer Institute, Ludwig Maximilians University, ⁴Department of Molecular Infection Biology, Helmholtz Center for Infection Research

This study describes the microscopic monitoring of pneumococcus adherence to von Willebrand factor strings produced on the surface of differentiated human primary endothelial cells under shear stress in defined flow conditions. This protocol can be extended to detailed visualization of specific cell structures and quantification of bacteria by applying differential immunostaining procedures.

Developmental Biology

Development of a Larval Zebrafish Infection Model for Clostridioides difficile

Junkai Li¹, Can M. Ünal², Kazuhiko Namikawa¹, Michael Steinert^3,4,5, Reinhard W. Köster¹

¹Division of Cellular and Molecular Neurobiology, Zoological Institute, Technische Universität Braunschweig, ²Department of Molecular Biotechnology, Turkish-German University, ³Institut für Mikrobiologie, Technische Universität Braunschweig, ⁴Braunschweig Integrated Centre of Systems Biology, ⁵Helmholtz Centre for Infection Research

Presented here is a safe and effective method to infect zebrafish larvae with fluorescently labeled anaerobic C. difficile by microinjection and noninvasive microgavage.