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目的研究白介素2(IL-2)cDNA协同单纯疱疹病毒1型(HSV-1)gD核酸疫苗免疫对机体体液免疫和细胞免疫应答的影响,以及在HSV-1病毒角膜攻击时的保护效果。方法利用pcDNA3.1载体分别构建HSV-1糖蛋白D和IL-2的真核表达质粒pgD和pIL-2,体外鉴定其表达。动物实验分为pcDNA3.1空载体对照组、pgD单独免疫组和pgD+pIL-2联合免疫组3组,具体为通过肌肉免疫接种BALB/c小鼠3次,间隔2周,每次接种质粒100μg,第3次免疫后2周,用酶联免疫吸附试验(ELISA)检测抗体滴度并做亚型分析,利用3H-TdR掺入法进行Th细胞增殖实验,ELISA试剂盒检测Th细胞分泌的IL-2、IFN-γ和IL-10水平,耳廓肿胀实验检测迟发型超敏反应(DTH)反应;HSV-1病毒角膜攻击后,裂隙灯显微镜观察角膜上皮病变。结果与pgD单独免疫组相比,pIL-2的协同免疫可以显著提高IgG2a抗体滴度、Th细胞增殖反应和DTH反应,Th细胞分泌IL-2和IFN-γ水平也显著提高,IL-10水平明显下降。在动物保护实验中,pIL-2的协同免疫明显增强了pgD疫苗在病毒攻击时对角膜的保护效果。结论IL-2cDNA的协同免疫可以显著提高pgD核酸疫苗诱导的体液免疫和细胞免疫应答水平,增加核酸疫苗的免疫保护效果。
Objective To investigate the effects of interleukin 2 (IL-2) cDNA in combination with herpes simplex virus 1 (HSV-1) gD DNA vaccine on humoral and cellular immune responses and the protective effect of HSV-1 virus corneal challenge. Methods The eukaryotic expression plasmids pgD and pIL-2 of HSV-1 glycoprotein D and IL-2 were constructed respectively by pcDNA3.1 vector and identified in vitro. Animal experiments were divided into pcDNA3.1 empty vector control group, pgD single immunization group and pgD + pIL-2 combined immunization group three groups, specifically by intramuscular immunization of BALB / c mice 3 times at intervals of 2 weeks, each inoculation of plasmid 100μg, 2 weeks after the third immunization, the antibody titers were detected by enzyme-linked immunosorbent assay (ELISA) and subtypes were analyzed. Th cell proliferation assay was performed by 3H-TdR incorporation method. ELISA kit was used to detect the secretion of Th cells IL-2, IFN-|Ã and IL-10 levels were measured. The auricle swelling test was used to detect delayed-type hypersensitivity (DTH) response. After HSV-1 virus cornea attack, corneal epithelial lesions were observed with slit lamp microscope. Results Compared with pgD alone group, synergistic immunization with pIL-2 significantly increased IgG2a antibody titer, Th cell proliferative response and DTH response, and the levels of IL-2 and IFN-γ secreted by Th cells were also significantly increased. IL-10 level Significant decline. In animal protection experiments, pIL-2 co-immunization significantly enhanced the protection of the pgD vaccine against corneas during virus challenge. Conclusion The synergistic immunization with IL-2 cDNA can significantly increase the level of humoral and cellular immune response induced by pgD DNA vaccine and increase the immunoprotection effect of nucleic acid vaccine.