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Helmholtz Centre for Infection Research

9 ARTICLES PUBLISHED IN JoVE

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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 Gröbe 3, Andreas Jeron 2, Dunja Bruder 1,2, Sabine Stegemann-Koniszewski 1
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.

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Immunology and Infection

Study of Phagolysosome Biogenesis in Live Macrophages
Marc Bronietzki 1, Bahram Kasmapour 1, Maximiliano Gabriel Gutierrez 1,2
1Research Group Phagosome Biology, Helmholtz Centre for Infection Research, 2Division of Mycobacterial Research, National Institute for Medical Research

Study of Phagolysosome Biogenesis in Live Macrophages

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Medicine

Rapid In Vivo Assessment of Adjuvant's Cytotoxic T Lymphocytes Generation Capabilities for Vaccine Development
Darío Lirussi 1, Thomas Ebensen 1, Kai Schulze 1, Elena Reinhard 1, Stephanie Trittel 1, Peggy Riese 1, Blair Prochnow 1, Carlos A. Guzmán 1
1Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research

We present here an application for a standard immunological technique (CFSE stained OT-I proliferation) intended to rapidly monitor adjuvant-mediated cytotoxic T lymphocyte (CTL) generation in vivo. This fast estimation of CTL capacities is useful for the development of prophylactic vaccines against intracellular pathogens as well as therapeutic cancer vaccines.

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Biology

Micromanipulation Techniques Allowing Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators
Georgi Dimchev 1,2, Klemens Rottner 1,2
1Division of Molecular Cell Biology, Zoological Institute, Technische Universität Braunschweig, 2Department of Cell Biology, Helmholtz Centre for Infection Research

We describe how micro- and photomanipulation techniques such as FRAP and photoactivation enable the determination of motility parameters and the spatiotemporal dynamics of proteins within migrating cells. Experimental readouts include subcellular dynamics and turnover of motility regulators or of the underlying actin cytoskeleton.

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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 Bergmann 1
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.

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Developmental Biology

Development of a Larval Zebrafish Infection Model for Clostridioides difficile
Junkai Li 1, Can M. Ünal 2, Kazuhiko Namikawa 1, Michael Steinert 3,4,5, Reinhard W. Köster 1
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.

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Immunology and Infection

P. aeruginosa Infected 3D Co-Culture of Bronchial Epithelial Cells and Macrophages at Air-Liquid Interface for Preclinical Evaluation of Anti-Infectives
Carlos Victor Montefusco-Pereira *1,2, Justus C. Horstmann *1,2, Thomas Ebensen 3, Christoph Beisswenger 4, Robert Bals 4, Carlos A. Guzmán 3, Nicole Schneider-Daum 1, Cristiane de Souza Carvalho-Wodarz 1, Claus-Michael Lehr 1,2
1Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), 2Department of Pharmacy, Saarland University, 3Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, 4Department of Internal Medicine V - Pulmonology, Allergology, Respiratory Intensive Care Medicine, Saarland University Hospital

We describe a protocol for a three-dimensional co-culture model of infected airways, using CFBE41o- cells, THP-1 macrophages, and Pseudomonas aeruginosa, established at the air-liquid interface. This model provides a new platform to simultaneously test antibiotic efficacy, epithelial barrier function, and inflammatory markers.

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Cancer Research

Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo
Julia Volckmar 1,2, Laura Knop 1,2,3, Tatjana Hirsch 1, Sarah Frentzel 1,2, Christian Erck 4, Marco van Ham 4, Sabine Stegemann-Koniszewski *1,2,5, Dunja Bruder *1,2
1Immune Regulation Group, Helmholtz Centre for Infection Research, 2Infection Immunology Group, Institute of Medical Microbiology, Infection Prevention and Control, Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke University Magdeburg, 3Institute for Molecular and Clinical Immunology, Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke University Magdeburg, 4Cellular Proteome Research, Helmholtz Centre for Infection Research, 5Experimental Pneumology, University Hospital of Pneumology, Health Campus Immunology, Infectiology and Inflammation, Otto-von-Guericke University Magdeburg

We describe a protocol for the chemical conjugation of the model antigen ovalbumin to an endocytosis receptor-specific antibody for in vivo dendritic cell targeting. The protocol includes purification of the antibody, chemical conjugation of the antigen, as well as purification of the conjugate and the verification of efficient conjugation.

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Immunology and Infection

Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test
Julia Häring *1, Tanja Michel *1, Matthias Becker 1, Daniel Junker 1, Tatia Tchitchagua 2, Olaf Leschnik 2, Berit Lange 3,4, Stefanie Castell 3,4, Gérard Krause 3,4, Monika Strengert 3, Alex Dulovic 1, Nicole Schneiderhan-Marra 1
1NMI Natural and Medical Sciences Institute at the University of Tübingen, 2Department of Neurology, Sächsisches Krankenhaus Rodewisch, 3Department of Epidemiology, Helmholtz Centre for Infection Research, 4German Centre for Infection Research (DZIF)

A three-channel dual-reporter fluorescence flow analysis system was used to develop a bead-based multiplex immunoassay that simultaneously evaluates serum samples for IgG and IgM elicited against multiple antigens of different Borrelia species that cause Lyme borreliosis in Europe and North America.

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