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J Vis Exp. 2019 Jun 23;(148). doi: 10.3791/59427.

Processing of Bronchoalveolar Lavage Fluid and Matched Blood for Alveolar Macrophage and CD4+ T-cell Immunophenotyping and HIV Reservoir Assessment.

Author information

1
Research Institute McGill University Health Centre; Department of Biological Sciences, Université de Québec à Montréal.
2
Research Institute McGill University Health Centre; Department of Biological Sciences, Université de Québec à Montréal; Department of Microbiology & Immunology, McGill University.
3
Department of Biological Sciences, Université de Québec à Montréal.
4
Centre de Recherche du Centre Hospitalier de l'Université de Montréal.
5
Centre de Recherche du Centre Hospitalier de l'Université de Montréal; Département de microbiologie, infectiologie et immunologie, Université de Montréal.
6
Research Institute McGill University Health Centre.
7
Research Institute McGill University Health Centre; Department of Microbiology & Immunology, McGill University; Department of Medicine, Division of Infectious Diseases, McGill University.
8
Department of Biological Sciences, Université de Québec à Montréal; Department of Microbiology & Immunology, McGill University; Département de microbiologie, infectiologie et immunologie, Université de Montréal; jenabian.mohammad-ali@uqam.ca.
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Contributed equally

Abstract

Bronchoscopy is a medical procedure whereby normal saline is injected into the lungs via a bronchoscope and then suction is applied, removing bronchoalveolar lavage (BAL) fluid. The BAL fluid is rich in cells and can thus provide a 'snapshot' of the pulmonary immune milieu. CD4 T cells are the best characterized HIV reservoirs, while there is strong evidence to suggest that tissue macrophages, including alveolar macrophages (AMs), also serve as viral reservoirs. However, much is still unknown about the role of AMs in the context of HIV reservoir establishment and maintenance. Therefore, developing a protocol for processing BAL fluid to obtain cells that may be used in virological and immunological assays to characterize and evaluate the cell populations and subsets within the lung is relevant for understanding the role of the lungs as HIV reservoirs. Herein, we describe such a protocol, employing standard techniques such as simple centrifugation and flow cytometry. The CD4 T cells and AMs may then be used for subsequent applications, including immunophenotyping and HIV DNA and RNA quantification.

PMID:
31282892
DOI:
10.3791/59427

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