摘要
目的 研究人单核细胞来源的树突状细胞 (DC)激活的HBsAg特异性细胞毒性T淋巴细胞 (CTL)对表达HBsAg的HepG2 /S靶细胞的杀伤效应 ,以探索DC HBsAg疫苗在抗乙型肝炎病毒 (HBV)中的作用。方法 从健康外周血中分离出单核细胞 ,在粒细胞 巨噬细胞集落刺激因子(GM CSF)和白细胞介素 4(IL 4)的作用下培养 7d诱导出DC ,然后以DC为刺激细胞在体外诱导出HBsAg特异性CTL ;将携有HBV S基因的pLXSN/S重组质粒电击导入肝癌细胞系 (HepG2 ) ,建立表达HBsAg的靶细胞模型HepG2 /S ;用染色法检测HBsAg特异性CTL对HepG2 /S靶细胞的杀伤效应。结果 DC激活的HBsAg特异性CTL对HepG2 /S靶细胞具有较强的杀伤效应 ,不同浓度HBsAg(0 μg/L ,5 0 μg/L和 10 0 μg/L)诱导的CTL的杀伤率分别为 3 .8%、69.5 %和85 .1% ,而CTL对HepG2细胞无明显杀伤效应。
Objective To study the HBsAg specific CTL activities activated by dendritic cells derived from human monocytes on the HepG2/S target cells, and further to probe into the anti HBV effect of HBsAg DC (dendritic cell) vaccine. Methods DCs are proliferated from human peripheral blood monocytes by adding GM CSF and IL 4 and then HBsAg specific CTL are activated by DCs pulsed by HBsAg; The target cell line (HepG2/S) expressing HBsAg was set up by transfecting recombinated plasmid with HBV/S gene (PLXSN/S) into HepG2/S cell line; HBsAg specific CTL and HepG2/S target cells were cocultured in 96 well flat bottomed microtiter plates for 48 hours at 37 °C in 5% CO 2, and then HBsAg specific CTL activities activated by DCs pulsed by HBsAg were detected by counting the number of killed target cells. Results HBsAg specific CTL activated by dendritic cells derived from human monocytes could produce strong killing effect on the target cells HepG2/S cells. It's specific CTL activities were 3.8%, 69.5% and 85.1% in different concentration (0 μg/L,50 μg/L and 100 μg/L) respectively. While, it had no killing effect on the HepG2 cells, so the HBsAg specific killing effect was specific. Conclusions The result shows that HBsAg specific CTL activated by dendritic cells derived from human monocytes has strong killing effect on the HBV.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2002年第6期333-336,共4页
Chinese Journal of Infectious Diseases
基金
国家自然科学基金资助项目 ( 39970 6 73)
