Centre de Recherche du Centre Hospitalier de l'Universite de Montreal (CRCHUM) 7 articles published in JoVE Biology Assessment of Gut Barrier Integrity in Mice Using Fluorescein-Isothiocyanate-Labeled Dextran Claire Gerkins1, Roy Hajjar1,2, Manon Oliero1, Manuela M. Santos1,3 1 In the present study, fluorescein-isothiocyanate-labeled (FITC) dextran is administered to mice via oral gavage to evaluate intestinal permeability both in vivo and in plasma and fecal samples. As gut barrier function is affected in many disease processes, this direct and quantitative assay can be used in diverse research areas. Medicine Large-Animal Model of Donation after Circulatory Death and Normothermic Regional Perfusion for Cardiac Assessment Khalil Khalil*1,2, Roberto V. P. Ribeiro*3, Julgans S. Alvarez4,5, Mitesh V. Badiwala4,5, Shant Der Sarkissian1,2, Nicolas Noiseux1,2 1 The protocol describes a large-animal (porcine) model of donation after circulatory death, followed by thoracoabdominal normothermic regional perfusion that closely simulates the clinical scenario in heart transplantation, and has the potential to facilitate therapeutic studies and strategies. Immunology and Infection Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment Manuel Flores Molina1,2, Thomas Fabre1,2, Aurélie Cleret-Buhot1, Geneviève Soucy1,3, Liliane Meunier1,4, Mohamed N. Abdelnabi1,2, Nicolas Belforte1,5, Simon Turcotte1,4,6, Naglaa H. Shoukry1,4,7 1 Here, we describe a simple and accessible strategy for visualizing, quantifying, and mapping immune cells in formalin-fixed paraffin-embedded tumor tissue sections. This methodology combines existing imaging and digital analysis techniques with the purpose of expanding the multiplexing capability and multiparameter analysis of imaging assays. Genetics Efficient Transcriptionally Controlled Plasmid Expression System for Investigation of the Stability of mRNA Transcripts in Primary Alveolar Epithelial Cells Francis Migneault1,2,3, Frédéric Gagnon2,3, Emmanuelle Brochiero1,3, Yves Berthiaume2,3, André Dagenais2,3 1 Here, we present a tool that can be used to study the posttranscriptional modulation of a transcript in primary alveolar epithelial cells by using an inducible expression system coupled to a pipette electroporation technique. Medicine Pre-clinical Model of Cardiac Donation after Circulatory Death Henry Aceros*1, Leyla Joulali*2, Mélanie Borie1, Roberto Vanin Pinto Ribeiro3, Mitesh Vallabh Badiwala3, Shant Der Sarkissian1,4, Nicolas Noiseux1,4 1 This protocol shows a simple and flexible approach for the evaluation of new conditioning agents or strategies to increase the feasibility of cardiac donation after circulatory death. Immunology and Infection Processing of Bronchoalveolar Lavage Fluid and Matched Blood for Alveolar Macrophage and CD4+ T-cell Immunophenotyping and HIV Reservoir Assessment Syim Salahuddin*1,2, Elaine Thomson*1,2,3, Oussama Méziane1,2, Omar Farnos2, Amélie Pagliuzza5, Nicolas Chomont5,6, Ron Olivenstein1, Cecilia Costiniuk1,3,4, Mohammad-Ali Jenabian2,3,6 1Research Institute McGill University Health Centre, 2Department of Biological Sciences, Université de Québec à Montréal, 3Department of Microbiology & Immunology, McGill University, 4Department of Medicine, Division of Infectious Diseases, McGill University, 5 We describe a method for processing bronchoalveolar lavage fluid and matched peripheral blood from chronically HIV-infected individuals on antiretroviral therapy to assess pulmonary HIV reservoirs. These methods result in the acquisition of highly pure CD4 T cells and alveolar macrophages that may subsequently be used for immunophenotyping and HIV DNA/RNA quantifications by ultrasensitive polymerase chain reaction. Immunology and Infection In Vitro Assay to Evaluate the Impact of Immunoregulatory Pathways on HIV-specific CD4 T Cell Effector Function Filippos Porichis*1, Meghan G. Hart*1, Jennifer Zupkosky1, Lucie Barblu1,2, Daniel E. Kaufmann1,2 1The Ragon Institute of MGH, MIT and Harvard, 2 We developed an in vitro assay to investigate the role of immunoregulatory pathways in the regulation of cytokine secretion by HIV-specific CD4 T cells.