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Processing of Bronchoalveolar Lavage Fluid and Matched Blood for Alveolar Macrophage and CD4+ T-cell Immunophenotyping and HIV Reservoir Assessment
- Source :
- Journal of Visualized Experiments.
- Publication Year :
- 2019
- Publisher :
- MyJove Corporation, 2019.
-
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.
- Subjects :
- 0301 basic medicine
Disease reservoir
Lung
General Immunology and Microbiology
medicine.diagnostic_test
business.industry
General Chemical Engineering
General Neuroscience
Context (language use)
respiratory system
General Biochemistry, Genetics and Molecular Biology
Flow cytometry
03 medical and health sciences
030104 developmental biology
0302 clinical medicine
Bronchoalveolar lavage
Immunophenotyping
Immune system
medicine.anatomical_structure
Immunology
medicine
Alveolar macrophage
business
030215 immunology
Subjects
Details
- ISSN :
- 1940087X
- Database :
- OpenAIRE
- Journal :
- Journal of Visualized Experiments
- Accession number :
- edsair.doi...........cb7d00a2750c6a95e2ce3b6589476fc0