Influence of extracellular signal-regulated kinase 1/2 pathway inhibitor PD98059 on proliferation and killing function of γδT cells cultured in vitro
10.3969/j.issn.1001-5256.2016.07.038
- VernacularTitle:细胞外信号调节激酶1/2通路抑制剂PD98059对体外培养γδT细胞增殖和杀伤功能的影响
- Author:
Leiqing SUN
1
;
Jing XU
;
Yongqiang CHEN
Author Information
1. Department of Central Laboratory, The 97th Hospital of PLA, Xuzhou, Jiangsu 221004, China
- Publication Type:Research Article
- Keywords:
extracellular signal-regulated MAP kinases;
protein kinase inhibitors;
receptors, antigen, T-cell, gamma-delta;
cell proliferation
- From:
Journal of Clinical Hepatology
2016;32(7):1388-1391
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of extracellular signal-regulated kinase 1/2 (ERK1/2) pathway inhibitor PD98059 on the proliferation and killing function of γδT cells cultured in vitro. MethodsMononuclear cells were separated from peripheral blood in healthy subjects and placed in RPMI 1640 complete medium containing zoledronic acid and interleukin-2 to obtain γδT cells through induction and culture. The ERK1/2 specific inhibitor PD98059 was used to block the ERK1/2 signaling pathway in γδT cells, and the proliferation of γδT cells was measured by cell counting, and the killing function of γδT cells was measured by CCK-8 method. Flow cytometry was used to measure the expression of granzyme B and perforin in γδT cells. The t-test was used for comparison of continuous data between groups. ResultsAfter 10 days of culture, the purity of γδT cells reached 87.94%±2.36%. The number of γδT cells treated with PD98059 was significantly lower than that of control cells ［(6.74±0.36)×105/ml vs (9.42±0.31)×105/ml, t=-12.708, P＜0.001］. Compared with the control cells, those treated with PD98059 had significantly higher positive expression rates of granzyme B and perforin (granzyme B: 48.89%±1.31% vs 41.58%±1.58%, t=7.582, P＜0.001; perforin: 65.92%±3.29% vs 33.49%±2.83%, t=15.478, P＜0001) and a significantly higher killing rate of HepG2 cells (69.28%±4.96% vs 48.34%±3.01%, t=11.201, P＜0.001). ConclusionThe ERK1/2 specific inhibitor PD98059 can inhibit the proliferation of γδT cells, but it can enhance the in vitro killing function of γδT cells.
