异戊烯焦磷酸刺激γδT细胞增殖及体外抗HBV作用  

Stimulation of gamma delta T cell by isopentenyl pyrophosphate and its effect of anti-HBV in vitro

作　　者：桂万羊[1,2] 马勇 陈复兴[3] 贺潇[1,2] 

机构地区：[1]蚌埠医学院医学检验系,安徽蚌埠233030 [2]解放军第一二三医院肝病中心,安徽蚌埠233015 [3]解放军第九七医院实验科,江苏徐州221004

出　　处：《临床肝胆病杂志》2008年第5期334-337,共4页Journal of Clinical Hepatology

摘　　要：目的采用异戊烯焦磷酸(isopentenyl pyrophosphate,IPP)刺激人外周血单个核细胞(peripheral bloodmononuclear cells,PBMCs)中γδT细胞特异性增殖,观察其体外对HBV复制和表达的抑制作用。方法体外用IPP和rhIL-2刺激人PBMCs培养10天,流式细胞术检测培养前后γδT细胞含量;将γδT细胞与HepG2.2.15细胞按一定比例共孵育培养后,ELISA法测定上清中HBsAg,HBeAg的表达,荧光定量PCR检测HBVDNA的含量。结果采用IPP和rhIL-2刺激人PBMCs,可以使人外周血γδT细胞选择性激活和扩增,由4.34%±1.79%。增加至55.65%±6.88%。所扩增的γδT细胞,能够部分抑制HepG2.2.15细胞中HBVDNA复制和HBeAg的表达,对HB-sAg的表达没有明显抑制作用。结论采用IPP刺激PBMCs,能使γδT细胞选择性增殖,增殖的细胞能够降低HepG2.2.15细胞中HBV的复制和HBeAg表达,在乙型肝炎的过继性免疫治疗中具有潜在的临床应用价值。Objective To investigate the inhibitory effect of peripheral blood mononuclear cells （PBMCs） stimulated by IPP on hepatitis B virus （HBV） in vitro. Methods The peripheral blood mononuclear cells were co - cultured with isopentenyl pyrophosphate （IPP） and recombinate human IL- 2 （rhlL- 2） as stimulating reagents in vitro. The expansion of gamma delta T cells was analysed by flowcytometry. PBMCs stimulated by IPP were added to HepG2. 2. 15 ceils in special ratio. HBsAg and HBeAg was measured by ELISA and HBV DNA was detected by fluorogenic quantitative PCR method in the supernatant. Results Using IPP stimulating PBMCs, gamma delta T cells were selected expansion from 4. 34% ± 1.79% to 55.65% ±6. 88%. Activated gamma delta T cells significantly reduced HBeAg secretion and HBV DNA replication of HepG2. 2. 15 cells. But having no remarkable inhibitory effects on HBsAg secretion of HepG2. 2. 15 cells. Conclusions PBMCs were stimulated by IPP and gamma delta T cells were selected proliferation, which can reduce the duplicate of HBV and express of HBeAg in HepG2. 2. 15 cells. Gamma delta T cells are of value in HBV adoptive immunotherapy.

关 键 词：异戊烯焦磷酸单酯 乙型肝炎 ΓΔT细胞 HEPG2.2.15细胞 

分 类 号：R512.62[医药卫生—内科学]