单纯疱疹病毒I型糖蛋白B基因疫苗免疫效果的初步研究
发布时间：2018-11-24 10:50
【摘要】： 目的:通过观察三种不同浓度的单纯疱疹病毒-I型糖蛋白B基因疫苗的免疫效果,探讨该疫苗在小鼠体内最佳免疫接种剂量,为核酸疫苗的应用奠定理论基础。 方法:昆明小鼠随机分为4组,分别接种pcDNA_3-gB_1 (200μg/0.1ml), pcDNA_3-gB_2(100μg/0.1ml), pcDNA_3-gB_3 (50μg/0.1ml)和pcDNA_3 (200μg/ 0.1ml),每次间隔2周,共免疫3次。在初次免疫前及免疫后的第7d, 21d, 35d,42d,49d,56d收集免疫鼠的血清,用间接ELISA法检测血清特异性抗体的变化;收集初次免疫后42天的脾细胞,用3H-TdR掺入法进行T细胞特异性抗原刺激实验;在第3次免疫后2周,用HSV-1SM44株病毒攻击小鼠角膜,检测疫苗接种对小鼠的保护效果。 结果:血清特异性抗体含量及T细胞刺激指数在各实验组(DNA疫苗接种组)中均高于空载体质粒pcDNA_3接种组,差异有统计学意义(P0.05);尤其是pcDNA_3-gB_1和pcDNA_3-gB_2组升高最为明显,但二者之间的免疫效果比较差异不明显,统计学上无显著性(P0.05)。在动物保护实验中,pcDNA_3-gB_1和pcDNA_3-gB_2组在病毒攻击时表现出明显的保护效果。 结论:pcDNA_3-gB基因疫苗免疫接种后可同时诱导机体产生特异性的体液免疫和细胞免疫,并具有免疫保护作用;不同浓度的核酸疫苗免疫效果不同,以高浓度效果为好;100μg/0.1ml的浓度为本疫苗在小鼠体内接种较为理想的剂量。
[Abstract]:Objective: to investigate the optimal dose of herpes simplex virus type I glycoprotein B gene vaccine in mice by observing the immune effect of three kinds of herpes simplex virus type I glycoprotein B gene vaccine in order to lay a theoretical foundation for the application of nucleic acid vaccine. Methods: Kunming mice were randomly divided into four groups: pcDNA_3-gB_1 (200 渭 g/0.1ml), pcDNA_3-gB_2 (100 渭 g/0.1ml), pcDNA_3-gB_3 (50 渭 g/0.1ml) and pcDNA_3 (200 渭 g / 0.1ml). Each time interval 2 weeks, a total of 3 times. The serum of immunized mice was collected on the 7th, 21st, 35d, 42nd, 49d and 56d before and after the first immunization, and the changes of serum specific antibodies were detected by indirect ELISA assay. Spleen cells were collected 42 days after the first immunization, T cell specific antigen stimulation test was carried out by 3H-TdR incorporation method, and HSV-1SM44 strain virus was used to attack the cornea of mice 2 weeks after the third immunization, and the protective effect of vaccine inoculation on mice was tested. Results: the content of serum specific antibody and T cell stimulating index in each experimental group (DNA vaccination group) were higher than those in pcDNA_3 inoculation group (P0.05). Especially in pcDNA_3-gB_1 and pcDNA_3-gB_2 group, the increase was most obvious, but there was no significant difference in the immune effect between the two groups (P0.05). In animal protection experiment, pcDNA_3-gB_1 and pcDNA_3-gB_2 groups showed obvious protective effect during virus attack. Conclusion: pcDNA_3-gB gene vaccine can induce specific humoral and cellular immunity at the same time. The concentration of 100 渭 g/0.1ml was the optimal dose of the vaccine in mice.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392
本文编号：2353404
[Abstract]:Objective: to investigate the optimal dose of herpes simplex virus type I glycoprotein B gene vaccine in mice by observing the immune effect of three kinds of herpes simplex virus type I glycoprotein B gene vaccine in order to lay a theoretical foundation for the application of nucleic acid vaccine. Methods: Kunming mice were randomly divided into four groups: pcDNA_3-gB_1 (200 渭 g/0.1ml), pcDNA_3-gB_2 (100 渭 g/0.1ml), pcDNA_3-gB_3 (50 渭 g/0.1ml) and pcDNA_3 (200 渭 g / 0.1ml). Each time interval 2 weeks, a total of 3 times. The serum of immunized mice was collected on the 7th, 21st, 35d, 42nd, 49d and 56d before and after the first immunization, and the changes of serum specific antibodies were detected by indirect ELISA assay. Spleen cells were collected 42 days after the first immunization, T cell specific antigen stimulation test was carried out by 3H-TdR incorporation method, and HSV-1SM44 strain virus was used to attack the cornea of mice 2 weeks after the third immunization, and the protective effect of vaccine inoculation on mice was tested. Results: the content of serum specific antibody and T cell stimulating index in each experimental group (DNA vaccination group) were higher than those in pcDNA_3 inoculation group (P0.05). Especially in pcDNA_3-gB_1 and pcDNA_3-gB_2 group, the increase was most obvious, but there was no significant difference in the immune effect between the two groups (P0.05). In animal protection experiment, pcDNA_3-gB_1 and pcDNA_3-gB_2 groups showed obvious protective effect during virus attack. Conclusion: pcDNA_3-gB gene vaccine can induce specific humoral and cellular immunity at the same time. The concentration of 100 渭 g/0.1ml was the optimal dose of the vaccine in mice.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392
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