Dear Colleagues
We have been looking at the expression of NKR; CD44, CD46, and NKG2A receptors on NK cells in PBMCs from normal donors both unstimulated and stimulated with IL-2 +IL-15, but unfortunately no increase in expression of these markers was measured! Could you please help us with the right protocol.
Secondly, we also measured very little intracellular expression of granzyme B and perforin in stimulated NK cells from PBMCs; is that what usually seen by others and how to increase the production of these molecules? What is the kinetics and the optimum time of incubation ?.
Hi Andre, Thanks for your advice ; yes we used IL-2 alone but not IL-15. but what is the incubation period?
I believe these articles may help you. Take a look!
http://online.liebertpub.com/doi/abs/10.1089/107999004323065093
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3085179/
Hi
What are your working concentrations?
I've been getting good results with 300U/ml
For how long have you cultured the cells?
I've been observing phenotypic chances after 36h or so. (haven't checked earlier).
have you tried enriching primary NK cells from whole blood?
I find negative selection methods to be more effective in the aspect of receptor modulation.
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