病毒灭活血浆对人CIK细胞功能影响的体外实验研究

In vitro study of the effect of virus inactivated plasma on the function of human CIK cells

目的利用CIK细胞(细胞因子诱导的杀伤细胞)来研究亚甲蓝光化学法病毒灭活血浆是否会对免疫细胞产生影响。方法对10名健康献血者外周血单个核细胞分别采用病毒灭活血浆和新鲜冰冻血浆培养人CIK细胞,观察2组培养体系中CIK细胞的扩增情况;用流式细胞仪(FCM)检测CIK细胞CD3+CD56+表达;检测经白藜芦醇终浓度为0.8μmol/L诱导48 h后的CIK细胞穿孔素和颗粒酶B的含量变化;以及用乳酸脱氢酶法测定CIK细胞杀伤SGC-7901细胞活性。结果新鲜冰冻血浆与病毒灭活血浆比较培养5、10、15 d CIK细胞的增殖倍数,分别为17.62±1.88、26.31±1.95、46.05±2.86与18.10±1.73、25.97±1.55、45.82±1.15,CD3+CD56+的表达分别为(12.37±1.38)%、(17.39±2.81)%、(24.3±1.72)%与(11.46±1.35)%、(18.36±1.96)%、(24.08±2.21)%,在促进CIK细胞的增殖以及CD3+CD56+的表达上,2组无统计学意义(P>0.05);经白藜芦醇终浓度为0.8μmol/L诱导培养48 h后,新鲜冰冻血浆与病毒灭活血浆培养的CIK细胞穿孔素、颗粒酶B、杀伤活性分别为(37.17±1.95)%、(38.79±1.91)%、(46.05±2.86)%和(32.04±1.92)%、(33.50±1.17)%、(45.82±1.15)%,2组无统计学意义(P>0.05)。结论病毒灭活血浆对人CIK细胞的增殖、细胞穿孔素和颗粒酶B含量变化以及体外杀伤功能均无明显影响。

Objective To study whether the immune cells could be affected by the virus inactivated plasma,which u-sing methylene blue hght chemical method,via CIK ceUs. Methods The peripheral mononuelear cells of 10 healthy blood donors were used to culture human CIK by using virus inactivated plasma and fresh frozen plasma. And the amplification of CIK cells was observed in the two groups. The CD3 ^＋ CD56^＋ expression on CIK ceils was detected via flow cytometry（FCM）.The variation of perforin and granzyme B content, which were induced by Resveratrol with a final concentration of 0.8 μmol/L for 48 hours were also detected. LDH assay was used to measure the CIK cell killing activity of SCG7901 cells. Re-suits The proliferation multiples of CIK cell, cultured by fresh frozen plasma and virus inactivated plasma at 5,10 and 15 days, were 17.62 ± 1.88,26.31 ± 1.95,46.05 ± 2.86, and 18.10 ± 1.73,25.97 ± 1.55,45.82 ± 1.15, respectively. The expression of CD3^＋ CD56^＋ and the promotion of CIK cell increment showed no significant difference between the two groups （ P 〉 0.05）. The perforin, granzyme B and killing activity of CIK cells in the fresh frozen plasma group and virus inactivated plasma group were（37.17 ± 1.95）%, （38.79 ± 1.91 ）%, （46.05 ± 2.86）% ,and （32.04 ± 1.92）%, （33.50 ± 1. 17 ） %, （45.82 ± 1.15 ） %, respectively. There was also no significant difference between 2 groups （ P 〉 0.05 ）. Conclusion Vires inactivated plasma had no effect on CIK cell proliferation, perforin, Granzyme B content and killing activity in vitro.