人巨细胞病毒蛋白pp65诱导特异性免疫应答的研究

目的研究人巨细胞病毒(human cytomegalovirus,HCMV)蛋白pp65(HCMV pp65)蛋白对特异性免疫应答的诱导作用。方法采用T细胞体外激活试验检测重组HCMVpp65蛋白对人外周血单个核细胞(perpheral blood mononuclear cells,PBMC)的刺激作用,并对激活的淋巴细胞进行传代培养,观察细胞的增殖;收集传代增殖的淋巴细胞,检测其对受HCMV感染的人胚肺成纤维细胞(human embryo lung fibro-blasts,HEL)的杀伤作用和对受感染后不同时段的HEL的杀伤功能。结果重组HCMV pp65蛋白在体外能激活PBMC,并使激活的细胞增殖;增殖的淋巴细胞可特异性地杀伤受HCMV感染的HEL细胞,但对未受感染的HEL细胞无杀伤作用,且其对受染2h后的HEL即有杀伤作用。结论重组HCMV pp65蛋白可诱导CD8 T细胞的激活与增殖,对受HCMV感染细胞产生特异性的杀伤作用,此作用不需要病毒转录和蛋白合成,对于HCMV的免疫治疗将有重要的作用。

Human Cytomegalovirus pp65 Protein Induces Specific Immune Response

Objective To investigate if recombinant HCMVpp65 can induce specific immune response in T cells.Methods The stimulating effect of recombinant HCMV pp65 protein on PBMC were observed in vitro. Cytotoxicity of PBMC (stimulated by rhHCMVpp65 protein) was detected by using PBMCs incubated with K562 cells or HCMV-infected human embryo lung fibroblasts (HEL) respectively, as well as HEL cells infected at various time points.Results rhHCMV pp65 protein could activate PBMCs and cause PBMC proliferation in vitro. The proliferating PBMCs had specific cytotoxicity to HEL cells infected by HCMV, but not to K562 cells and uninfected HEL cells. CTL could kill the HEL cells 2 h after it was infected with HCMV, and its cytotoxicity persisted for 68 h.Conclusion Recombinant HCMV pp65 protein could induce specific immune response in T cells in vitro. CTL could kill the HCMV-infected HEL cells, and the cytotoxicity of CTL is independent of viral gene expression and requires no protein synthesis. It is important to the immunotherapy of HCMV infection.