Protocol for Analyzing Mycobacteria-Specific Cells in Human Airways
Author Information
Author(s): Kiravu Agano, Rozot Virgine, Cruywagen Lauren, Gutschmidt Andrea, DuPlessis Nelita, Nemes Elisa, Tameris Michele, Scriba Thomas, Conradie Arina, Kafaar Fazlin, van Rensburg Ilana C., Walzl Gerhard, Malherbe Stephanus, Shabangu Ayanda, Middelkoop Keren
Primary Institution: University of Cape Town
Conclusion
The protocol allows for stable measurement of myeloid cell markers after freezing, enabling efficient analysis of human airway responses to mycobacterial antigens.
Supporting Evidence
- The protocol includes steps for thawing and stimulating cryopreserved bronchoalveolar lavage cells.
- Markers of myeloid lineage and function remain stable after freezing.
- An optimized antibody panel for mass cytometry is provided.
Takeaway
This study shows how to prepare and analyze immune cells from the lungs to see how they react to certain bacteria, using a special method that keeps the cells stable even after freezing.
Methodology
The study outlines a protocol for in vitro stimulation of cryopreserved bronchoalveolar lavage cells with mycobacterial antigens and subsequent analysis using mass cytometry.
Potential Biases
There may be risks of bias due to variations in sample handling and processing conditions.
Limitations
The protocol is limited by the number of samples that can be processed and the potential for contamination affecting results.
Participant Demographics
Participants were healthy individuals who tested negative for HIV, SARS-CoV2, and Mycobacterium tuberculosis.
Digital Object Identifier (DOI)
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