单纯疱疹病毒2型gD蛋白-结核杆菌热休克蛋白70 DNA疫苗的免疫效应研究  

Immune response induced by Hsp70-HSV2gD DNA vaccine

作　　者：樊建勇 田晨 王兴旺 张功恺 高睿 籍丽玥 陈韦君 杨慧兰 FAN Jian-yong;TIAN Chen;WANG Xing-wang(Department of Dermatology,General Hospital of Southern Theatre Command of PLA,Guangzhou 510010,China)

机构地区：[1]南部战区总医院皮肤科,广州510010

出　　处：《实用皮肤病学杂志》2021年第1期8-11,共4页Journal of Practical Dermatology

基　　金：国家自然科学基金资助项目(81071401)。

摘　　要：目的研究单纯疱疹病毒2型gD蛋白-结核杆菌热休克蛋白70(Hsp70-HSV2gD)DNA疫苗诱导BALB/c小鼠出现的特异性免疫反应.方法用制备的Hsp70-HSV2gD DNA疫苗免疫小鼠3次,最后1次免疫后第4周测定小鼠血中特异性gD IgG抗体;分离小鼠脾淋巴细胞,用HSV2gD蛋白刺激后用MTT法测定小鼠脾淋巴细胞增殖水平,用酶联免疫吸附试验(ELISA)测定脾淋巴细胞培养液中干扰素(IFN)-γ、白细胞介素(IL)-4浓度,用流式细胞仪检测小鼠脾淋巴细胞CD4+、CD8+T细胞亚群.结果gD、gD+Hsp70和Hsp70-HSV2gD 3组均可刺激小鼠产生靶向HSV2gD蛋白抗体,诱导CD4+T细胞与CD8+T细胞增殖,刺激脾淋巴细胞增殖并分泌IFN-γ、IL-4.Hsp70-HSV2gD组在抗体水平、诱导CD4+T细胞与CD8+T细胞增殖、刺激脾淋巴细胞增殖及分泌IFN-γ、IL-4均明显高于gD+Hsp70组、gD组,差异有统计学意义(P<0.05).结论Hsp70-HSV2gD DNA疫苗可以刺激小鼠产生比gD、gD+Hsp70组更强的特异性免疫反应,Hsp70-HSV2gD DNA疫苗中的Hsp70对HSV2gD抗原诱导的特异性免疫应答可能有促进作用.Objective To evaluated the specific humoral and cellular immune responses induced by Hsp70-HSV2gD DNA vaccine in BALB/c mice.Methods The Hsp70-HSV2gD DNA vaccine was used to inoculate BALB/c mice for three times.Four weeks after the last vaccination,the level of anti-HSV-2gD IgG antibody was detected.At the same time,the spleen cells of the vaccinated mice were separated and stimulated with HSV-2gD protein and the proliferation of the spleen lym phocytes was detected by MTT assay;the percentage of CD4^(+) T cells and CD8^(+)T cells was detected by flow cytometry(FCM)and the level of IFN-γ and IL-4 in culture was detected by ELISA assay.Results After induced by Hsp70-HSV2gD,gD and gD+Hsp70 DNA vaccine respectively,the BALB/c mice could produce the antibody against HSV-2gD.the percentage of CD4^(+)T cells and CD8^(+)T cells increased,and the splenocytes proliferated and secreted IFN-γ and IL-4.All the testing indexes mentioned above of Hsp70-HSV2gD DNA vaccine group were obviously higher than the other control groups(P<0.05).Conclusion The Hsp70-HSV2gD DNA vaccine could induce higher immune response than that of gD and gD+Hsp70 DNA vaccine,which suggests that the adjuvant Hsp70 of Hsp70-HSV2gD DNA vaccine maybe promote specific immune response.

关 键 词：病毒 单纯疱疹 热休克蛋白70 DNA疫苗 

分 类 号：R392.7[医药卫生—免疫学] R752.11[医药卫生—基础医学]