German Centre for Infection Research (DZIF) 6 articles published in JoVE Immunology and Infection Simultaneous Detection of Different Antibody Classes in a Multiplexed Serological Test Julia Häring*1, Tanja Michel*1, Matthias Becker1, Daniel Junker1, Tatia Tchitchagua2, Olaf Leschnik2, Berit Lange3,4, Stefanie Castell3,4, Gérard Krause3,4, Monika Strengert3, Alex Dulovic1, Nicole Schneiderhan-Marra1 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. Bioengineering Metabolic Profiling to Determine Bactericidal or Bacteriostatic Effects of New Natural Products using Isothermal Microcalorimetry Katarina Cirnski*1,2, Janetta Coetzee*1,2, Jennifer Herrmann1,2, Rolf Müller1,2 1Helmholtz Institute for Pharmaceutical Research Saarland, Department of Microbial Natural Products, Helmholtz Centre for Infection Research and Department of Pharmacy, Saarland University, 2Partner Site Hannover-Braunschweig, German Centre for Infection Research (DZIF) The elucidation of the mode-of-action of a novel antibiotic is a challenging task in the drug discovery process. The goal of the method described here is the application of isothermal microcalorimetry using calScreener in antibacterial profiling to provide additional insight into drug-microbe interactions. Immunology and Infection CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites Julia K. Bialek*1,2, Thomas Walther*1, Joachim Hauber1,3, Ulrike C. Lange1,2,3 1Heinrich Pette Institute, Leibniz Institute for Experimental Virology, 2Department of Anesthesiology, University Medical Center Hamburg-Eppendorf, 3German Center for Infection Research (DZIF) We present a genome engineering workflow for the generation of new in vitro models for HIV-1 infection that recapitulate proviral integration at selected genomic sites. Targeting of HIV-derived reporters is facilitated by CRISPR-Cas9-mediated, site-specific genome manipulation. Detailed protocols for single-cell clone generation, screening, and correct targeting verification are provided. Immunology and Infection Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection Thomas Tu1,2, Stephan Urban1,3 1Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 2Gastroenterology and Liver Research Laboratory, Ingham Institute, 3German Center for Infection Research (DZIF) We describe here the in vitro generation of HBV DNA via a Hepatitis B virus infection system and the highly sensitive detection of its (1–2 copies) integration using inverse nested PCR. Biology Forced Salivation As a Method to Analyze Vector Competence of Mosquitoes Anna Heitmann*1, Stephanie Jansen*1,2, Renke Lühken1, Mayke Leggewie1,2, Jonas Schmidt-Chanasit1,2, Egbert Tannich1,2 1Bernhard Nocht Institute for Tropical Medicine, 2German Centre for Infection Research (DZIF) For efficient control of mosquito borne virus transmission, the knowledge of the vector potential of respective mosquitoes is of particular interest. We describe forced salivation as a method to analyze vector competence of Aedes albopictus and three different Culex taxa for the transmission of Zika virus. Medicine In Vivo Tracking of Edema Development and Microvascular Pathology in a Model of Experimental Cerebral Malaria Using Magnetic Resonance Imaging Angelika Hoffmann1,2, Xavier Helluy2,3, Manuel Fischer2, Ann-Kristin Mueller4,5, Sabine Heiland2, Mirko Pham1,6, Martin Bendszus1, Johannes Pfeil4,5,7 1Department of Neuroradiology, Heidelberg University Hospital, 2Division of Experimental Radiology, Department of Neuroradiology, Heidelberg University Hospital, 3NeuroImaging Centre Research, Department of Neuroscience, Ruhr-University Bochum, 4Centre for Infectious Diseases, Parasitology Unit, Heidelberg University Hospital, 5German Centre for Infection Research (DZIF), 6Department of Neuroradiology, University of Würzburg, 7Center for Childhood and Adolescent Medicine, General Pediatrics, Heidelberg University Hospital We describe a mouse model of experimental cerebral malaria and show how inflammatory and microvascular pathology can be tracked in vivo using magnetic resonance imaging.
