【摘要】： 目的:通過觀察三種不同濃度的單純皰疹病毒-I型糖蛋白B基因疫苗的免疫效果,探討該疫苗在小鼠體內(nèi)最佳免疫接種劑量,為核酸疫苗的應(yīng)用奠定理論基礎(chǔ)。 方法:昆明小鼠隨機(jī)分為4組,分別接種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),每次間隔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,用間接ELISA法檢測血清特異性抗體的變化;收集初次免疫后42天的脾細(xì)胞,用3H-TdR摻入法進(jìn)行T細(xì)胞特異性抗原刺激實驗;在第3次免疫后2周,用HSV-1SM44株病毒攻擊小鼠角膜,檢測疫苗接種對小鼠的保護(hù)效果。 結(jié)果:血清特異性抗體含量及T細(xì)胞刺激指數(shù)在各實驗組(DNA疫苗接種組)中均高于空載體質(zhì)粒pcDNA_3接種組,差異有統(tǒng)計學(xué)意義(P0.05);尤其是pcDNA_3-gB_1和pcDNA_3-gB_2組升高最為明顯,但二者之間的免疫效果比較差異不明顯,統(tǒng)計學(xué)上無顯著性(P0.05)。在動物保護(hù)實驗中,pcDNA_3-gB_1和pcDNA_3-gB_2組在病毒攻擊時表現(xiàn)出明顯的保護(hù)效果。 結(jié)論:pcDNA_3-gB基因疫苗免疫接種后可同時誘導(dǎo)機(jī)體產(chǎn)生特異性的體液免疫和細(xì)胞免疫,并具有免疫保護(hù)作用;不同濃度的核酸疫苗免疫效果不同,以高濃度效果為好;100μg/0.1ml的濃度為本疫苗在小鼠體內(nèi)接種較為理想的劑量。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R392

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