Quantification of Inflammatory Mediators in Infected Human Tissue-Derived Cells
Published: September 29, 2023
Abstract
Source: Dousha, L. et al., Assessing Respiratory Immune Responses to Haemophilus Influenzae. J. Vis. Exp. (2021)
This video demonstrates the quantification of inflammatory mediators during infection by incubating human lung-tissue-derived immune cells with Haemophilus influenzae. Flow cytometry is used to measure cytokine fluorescence in labeled lymphocytes, correlating with the production of pro-inflammatory cytokines in response to bacterial infection.
Protocol
All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Antigenic preparation Live bacteria First, characterize live bacteria. Obtain the bacteria from the appropriate sample (e.g., sputum or bronchoscopy). Confirm the strain as Haemophilus influenzae using an appropr…
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The authors have nothing to disclose.
Materials
| Ammonium chloride | Sigma Aldrich | 213330 | |
| Brefeldin | Sigma Aldrich | B6542 | |
| Filcon sterile nylon mesh | Becton Dickinson | 340606 | |
| Gelatin substrate, Enzchek | Molecular probes | E12055 | |
| MACS mix tube rotater | Miltenyi Biotec | 130-090-753 | |
| Medimachine | Becton Dickinson | Catalogue number not available | |
| Saponin | Sigma Aldrich | 8047152 |
