Hey guys/girls, has anyone had experiencing using PMA/ionomycin to trigger Nk cell degranulation without using a target cell? such as a positive control for maximum degranulation or for a forced cytotoxic response from Nk cells?
Thank you all
Cord blood (CB) is increasingly used as a source of stem cells for hematopoietic stem cell transplantation, and natural killer (NK) cells may be the effectors of the antileukemic response observed after CB transplantation. Both phenotype and functions of CB NK cell subsets are determined by the percentage of NK cells which may be higher in CB compared with peripheral blood (PB). Furthermore, there occurs a higher percentage of the CD56bright subset in CB. CB NK cells reached a late stage of differentiation, but exhibited higher expression of NKG2A and expressed fewer killer-cell immunoglobulin-like receptors, suggesting an incomplete maturation. CB NK cells are highly expressed CXCR4, but did not express L-selectin, highlighting unique homing properties of CB NK cells. CB NK cells proliferated in response to interleukin-2 and degranulated in response to stimulation with tumor cells, but failed to lyse K562 cells in 51Cr-release assay. CB NK cells exhibited a lower interferon- production in comparison with PB NK cells. Culture with IL-2 increased CB NK cell functions.
Hi Lebaron,
yes, we use PMA/Iona as a positive control (for degranulation as well as cytokine production=IFNy/TNfa).
50 ng/mL PMA + 0,5 µM Ionomycin and 4h incubation should give you nice signals.
Good luck,
Markus
Agreed - PMA/Iono is our positive control of choice for both cytotoxicity and cytokine release.
-Andrew
Markus Granzin Do you used PMA only for nk cell degranulation? Do you have the mechanism? I am looking for a stimulus to generate expression of granzymes and perforins at the transcriptional and protein level in rainbow trout nk cells
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology