May I measure the CD107a marker on NK cells derived from patients treated with immunotherapy directly without in vitro stimulation with target cells or interleukins? I suppose that expression of CD107a was stimulated by immunotherapy in patient blood, and it does not require another stimulation. Am I right?
Thank you in advance!
Yes, you can, and you can see cd107a expression either without stimulation.
Please look at this reference: https://cancerimmunolres.aacrjournals.org/content/7/5/841
Good luck!
Yes, you can if you are interested in determining the frequency of degranulation immediately ex vivo. I have looked at NK cells immediately ex vivo from the blood and tumor tissue of patients with solid tumors, and the NK cells from the tumor have highly variable levels of CD107a (as you might expect), while NK cells from the blood generally have a low level, similar to what is seen from healthy donors. Hope that helps. I would add that you should stain for CD107a immediately after obtaining your sample of interest, as it is constantly recycled from the membrane.
Hi Adam M. Farkas :)
In up to how many minutes/hours the staining with anti-CD107a can be performed (before the decrease in CD107a expression on the cell surface)?
Would this apply to gamma-delta T cells? Thanks a lot! :)
Mateus Souza-Barros Hi Mateus!
Good question, I do not know exactly. However, I when I stain freshly-isolated PBMC or freshly-dissociated tumor tissue with CD107a (in the presence of Monensin), I can see different levels of ex vivo degranulation from various patients and healthy controls.
The monensin is very important as it should prevent recycling of CD107a back into the cytosol. For sure this method may not be 100% reflective of what's happening in vivo, but if samples are processed as consistently as possible after collection from the patient I think it's ok.
The patient samples I do this on are approximately 2h after collection I think, and it still works so a bit more time is likely ok as well (as long as kept constant).
I think you should be able to do this on gamma deltas as well, but I have not tried that personall. Best of luck.
Adam
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