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ISSN 1001-5256 (Print)
ISSN 2097-3497 (Online)
CN 22-1108/R
Issue 7
Jul.  2016
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Influence of extracellular signal-regulated kinase 1/2 pathway inhibitor PD98059 on proliferation and killing function of γδT cells cultured in vitro

DOI: 10.3969/j.issn.1001-5256.2016.07.038
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  • Published Date: 2016-07-20
  • Objective To 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. Methods Mononuclear 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. Results After 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 < 0. 001) and a significantly higher killing rate of Hep G2 cells( 69. 28% ± 4. 96% vs 48. 34% ± 3. 01%,t = 11. 201,P < 0. 001). Conclusion The ERK1/2specific inhibitor PD98059 can inhibit the proliferation of γδT cells,but it can enhance the in vitro killing function of γδT cells.

     

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