乙型肝炎病毒表面抗原对实体肿瘤患者CIK细胞诱导培养及表型分析的影响

Effects of hepatis B vines on the growth and cell subsets analysis of cytokine-induced killer cells(CIK) in vitro incubation from tumor

目的探讨乙型肝炎病毒表面抗原阳性患者与阴性患者细胞因子诱导的杀伤细胞(cytokine induced killer cells,CIK)体外培养过程中个细胞增殖情况和淋巴细胞亚群的变化。方法选取海军总医院2012年10月-2013年6月住院的肿瘤患者,共50例.根据乙型肝炎病毒表面抗原表达情况分为阴性组和阳性组。分别抽取患者外周血进行CIK细胞诱导培养,在第1、5、7、9、11、13、15天进行细胞活力检测并计数,第5、7、10、13、15天进行各细胞亚群的表达分析。结果两组患者CIK增殖情况单个时间点比较无显著性差异,但是在第15天的增殖倍数阳性组显著高于阴性组(280倍比180倍);细胞亚群分析中CD3^+T细胞、CD8^+T细胞、CD3^+CD8^+T细胞、CD3^+CD56^+T细胞比例均随培养时间延长而升高,CD4^+T细胞、CD3^+CD4^+T细胞随培养时间延长而降低,阳性组更显著;其中CD8~6T细胞和CD3^+CD4^+T细胞在两组间比较差异有统计学意义(P<0.05)。结论乙型肝炎病毒表面抗原阳性患者CIK细胞较阴性患者扩增能力更强,效应细胞表型表达更有利于杀伤作用的发挥。

Objective To explore the difference of the immune cell subsets during the process of cytokine-induced kill cells （CIK） incubation from tumor patients with and without hepati- tis B virus. Methods The peripheral blood was extracted from 50 tumor patients from Oct 2012 to Jun 2013 who were divided into two groups by with or without hepatitis B virus. The proliferate ve- loeity and activity of CIK cells were detected by count on 1,5,7,9,11,13 and 15 days of the CIK culture. The expressions of CD3＋, CD4＋, CD8＋, CD3＋ CD8＋, CD3＋ CD4＋, CD3＋ CD56＋ T cells were analyzed by flow cytometry on 5,7, 10,13 and 15 days of the culture. Results The proliferative multiple at 15 days in HBV positive group was higher than that of negative group （280 vs. 180）, al- though there was no significant difference between the two groups at the same culturing time points. The frequencies of CD3＋T cells, CDS＋T cells, CD3＋CD8＋T cells and CD3＋CD56＋T cells increased by the time, whilethe CD4＋T cells and CD3＋CD4＋T cells decreased by the time,the positive group was more than the negative. Among them, the difference of CD8＋T cells and CD3＋CD4＋T cells in two groups was significant （P〈0. 05）. Conclusion The proliferate multiple capacity of CIK cells from patients with HBV positive group was better than that of negative group, and the immune cell subsets states of HBV positive group was better than that of negative group.