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目的 构建人巨细胞病毒 pp6 5原核表达载体 ,研究纯化后的重组 pp6 5蛋白诱导特异性免疫应答的作用及其特异性。 方法 采用体外 DNA重组技术 ,构建 pp6 5原核表达载体并诱导其表达 ,通过亲和层析对表达产物进行纯化和经 Westen blot鉴定 ;用 MTT法检测重组蛋白对人外周血单个核细胞 (peripheral blood m ononuclear cell,PBMC)的激活、增殖作用 ,检测激活淋巴细胞对受 HCMV感染的人胚肺成纤维细胞 (human em bryo lung fibroblast,HEL )的杀伤作用。 结果 成功构建 pp6 5原核表达载体并诱导其高效表达 ,纯化后的 pp6 5蛋白能促进 PBMC激活和增殖 ,增殖的细胞可特异性地杀伤受 HCMV感染的 HEL细胞。 结论 经重组 pp6 5蛋白激活增殖的淋巴细胞 ,可特异性杀伤受 HCMV感染细胞 ,对 HCMV感染的免疫治疗具有十分重要的作用。
Objective To construct the prokaryotic expression vector of human cytomegalovirus (pp6 5) and study the role and specificity of the purified recombinant pp6 5 protein in inducing specific immune response. Methods The prokaryotic expression vector pp6 5 was constructed and its expression was induced by DNA recombination in vitro. The expressed product was purified by affinity chromatography and identified by Westen blot. The effect of recombinant protein on the expression of peripheral blood mononuclear cells (peripheral blood To investigate the cytotoxic effect of activated lymphocytes on HCMV-infected human embryonic lung fibroblasts (HEL). Results The prokaryotic expression vector pp6 5 was successfully constructed and induced to be highly expressed. The purified pp6 5 protein could promote the activation and proliferation of PBMC. The proliferating cells could specifically kill HEL cells infected with HCMV. Conclusion The lymphocytes activated by recombinant pp6 5 protein can specifically kill cells infected by HCMV and play an important role in the immunotherapy of HCMV infection.