Protocol to phenotype and quantify mycobacteria-specific myeloid cells from human airways by mass cytometry
Alveolar macrophages and other myeloid cells in the human airways are the primary cell types responding to respiratory pathogens. Here, we present a protocol for in vitro stimulation of cryopreserved human bronchoalveolar lavage (BAL) cells with mycobacterial antigens for phenotyping and quantifying proinflammatory cytokine responses in myeloid cells by mass cytometry. We demonstrate that the measure of markers of myeloid lineage and function is stable after freezing stained cells thereby allowing for batched analyses and/or machine downtime.
Authors
- Andrea Gutschmidt (ORCID: https://orcid.org/0000-0001-9231-8304)
- Elisa Nemes (ORCID: https://orcid.org/0000-0003-1662-4961)
- Virgine Rozot
- Agano Kiravu (ORCID: https://orcid.org/0000-0003-2510-2479)
- Lauren Cruywagen
- Nelita DuPlessis
Institutions
- South African Medical Research Council (ZA)
- University of Cape Town (ZA)
- Stellenbosch University (ZA)
- South African Tuberculosis Vaccine Initiative (ZA)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-12-01
- DOI
- https://doi.org/10.1016/j.xpro.2024.103463
- Primary Topic
- Mycobacterium research and diagnosis
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Israel Cancer Research Fund
- Universiteit Stellenbosch
- University of Cape Town
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases