Does anyone have a protocol for measuring T lymphocyte activation, either by flow cytometry or by measuring cytokine production? I'm working with CAR-T lymphocytes and I want to see if incubating them with patient sera will activate them in a specific manner. Can you suggest different cytokines and activation markers?
Hi Sergi Betriu Méndez
I briefly wanted to reach out concerning your inquiry.
For a detailed list of flow cytometry agents for the characterization of activation markers on the cell surface please consider the following paper from my old boss.
Article Advanced Flow Cytometry Assays for Immune Monitoring of CAR-...
For quick insights into activation markers for cytotoxic T cells and a basic flow cytometry protocol, please consider:
Article A role for the histone H2A deubiquitinase MYSM1 in maintenan...
For a comprehensive cytokine analysis (48plex with absolute quantification) use:
https://olink.com/products-services/target/48-cytokine-panel/
And for a detailed proteomic analysis and possible targets (using cell culture supernatants or serum (prior to incubation)) please consider the T96 INF, IR and IO panels (albeit with relative quantification of expression) as per:
https://olink.com/products-services/target/inflammation/
and more specifically used in the following papers:
Article Interaction kinetics with transcriptomic and secretory respo...
Article A phase I/IIa trial using CD19-targeted third-generation CAR...
I hope that helps.
All the best & good luck with your experiments,
Michael
I think this is all I need for now. I'll go through it thoroughly. Thanks a lot Michael Förster
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