Background LAIR-1 can be triggered by collagen and its intracellular ITIMs are crucial for the inhibitory signal to NK cells. In order to investigate the regulation mechanisms of LAIR-1/collagen on NK cells, we aimed to construct LAIR-1 over-expressed NK-92 cells with which STATs related analysis was performed.
Methods Lentivirus-mediated transfection was used to construct LAIR-1 over-expressed NK-92 cells. The expression of LAIR-1 in the constructed NK-92 cells was identified by QPCR and Western blot analysis. Immunofluorescence test was also used to observe the efficiency of lentivirus-mediated expression of LAIR-1 in NK-92 cells. The LAIR-1 over-expressed NK-92 cells were cultured in the IL-2 free medium for 48 hours and then different concentrations of IL-2 were added into the culture system and the cells were collected after 30 mins. The expression of STAT1, p-STAT1, STAT4, and p-STAT4 were analysed by Western blot analysis.
Results QPCR and Western blot analysis results showed the over-expressed LAIR-1 in NK-92 cells. Immunofluorescence result showed high efficiency of LAIR-1 lentiviral infection. Western blot results showed IL-2 could increase the phosphorylation of STAT1 in a dose-dependent manner, but not STAT4.
Conclusion LAIR-1 over-expressed NK-92 cells were constructed successfully and the IL-2 treatment experiment showed different roles of STAT1 and STAT4 in the regulation of NK cells.
Acknowledgements Supported by a project grant from the Natural Science Foundation of Shandong Province (ZR2015JL027) and National Natural Science Foundation of China (81370730, 81571512, 81273200, and 31300751).
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.