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2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Imaging InlC Secretion to Investigate Cellular Infection by the Bacterial Pathogen Listeria monocytogenes

Andreas Kühbacher^1,2,3, Edith Gouin^1,2,3, Jason Mercer⁴, Mario Emmenlauer⁵, Christoph Dehio⁵, Pascale Cossart^1,2,3, Javier Pizarro-Cerdá^1,2,3

¹Unité des Interactions Bactéries Cellules, Pasteur Institute, ²INSERM U604, ³Institut National de la Recherche Agronomique (INRA), USC2020, ⁴Institute of Biochemistry, ETH Zürich, ⁵Focal Area Infection Biology, Biozentrum, University of Basel

Listeria monocytogenes is a Gram positive bacterial pathogen frequently used as a major model for the study of intracellular parasitism. Imaging late L. monocytogenes infection stages within the context of small-interfering RNA screens allows for the global study of cellular pathways required for bacterial infection of target host cells.

Immunology and Infection

Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells

Alain Casanova^*1, Shyan H. Low^*1, Mario Emmenlauer^*1,4, Raquel Conde-Alvarez^*1,3, Suzana P. Salcedo², Jean-Pierre Gorvel², Christoph Dehio¹

¹Focal Area Infection Biology, Biozentrum, University of Basel, ²Centre d’Immunologie de Marseille-Luminy, Université de la Méditérannée UM2, INSERM U1104 CNRS UM7280, ³Departmento de Microbiologìa and Instituto de Salud Tropical, Universidad de Navarra, ⁴BioDataAnalysis GmbH

Two assays for microscopy-based high-throughput screening of host factors involved in Brucella infection are described. The entry assay detects host factors required for Brucella entry and the endpoint assay those required for intracellular replication. While applicable for alternative approaches, siRNA screening in HeLa cells is used to illustrate the protocols.