B型流感病毒核酸疫苗和CpG基序免疫增强作用研究

发布时间：2018-01-09 20:21

本文关键词：B型流感病毒核酸疫苗和CpG基序免疫增强作用研究　出处：《湖南师范大学》2005年硕士论文　论文类型：学位论文

【摘要】：本论文介绍了B型流感病毒DNA疫苗以及CpG基序作为佐剂对其免疫增强效应的研究。血凝素(Hemagglutinin,HA)和神经氨酸酶(neuraminidase,NA)是B型流感病毒两种最重要的抗原蛋白,本文首先探讨了一次接种B型流感病毒(B/Ibaraki/2/85)DNA疫苗对BALB/c小鼠的免疫保护作用。分别采用不同剂量的HA DNA疫苗(100μg、50μg、10μg、5μg、1μg),NA DNA疫苗(50μg、10μg、5μg、1μg),以及HA和NA联合DNA疫苗(10μg、5μg、1μg),一次接种BALB/c小鼠。接种后四周,用致死量流感病毒(B/Ibaraki/2/85)攻击小鼠,病毒攻击后三天,取血清检测抗-HA和抗-NA抗体。同时测定小鼠肺部病毒含量,观察记录小鼠存活率和体重丢失情况。结果发现:100μg HA DNA疫苗,10μg NA DNA疫苗以及5μg联合DNA疫苗(HA+NA)接种的小鼠全部存活。结合小鼠肺部病毒含量和体重变化数据,得出以下结论:一次接种DNA疫苗可以为小鼠提供完全有效的保护,联合DNA疫苗免疫保护作用强于单一DNA疫苗,NA DNA疫苗免疫保护作用略强于HA DNA疫苗。为了探讨CpG基序作为免疫佐剂的效应,本研究构建了包含CpG基序的质粒:pcD3d(+CpG)HA和不包含CpG基序的质粒pcD3d(-)HA。并采用不同的剂量,不同的免疫次数(一次或两次),分别免疫BALB/c小鼠,初免后四周(或加强免疫后一周),用致死量流感病毒(B/Ibaraki/2/85)攻击小鼠,病毒攻击后三天,取血清检测抗-HA IgG抗体,并观察小鼠存活率和体重变化情况。结果显示,含CpG基序的DNA疫苗能够进一步提高抗体水平,改善小鼠病毒攻击后的临床体症。实验表明CpG基序能有效提高小鼠抗B型流感病毒攻击的能力。
[Abstract]:In this paper, the immune enhancement effects of influenza B virus DNA vaccine and CpG motif as adjuvant were introduced. Hemagglutinin (Hemagglutinine HA) and neuraminidase (NAA) are two of the most important antigen proteins of influenza B virus. In this paper, we first looked at a single vaccination against influenza B virus B / Ibaraki / 2 / 85. The protective effect of DNA vaccine on BALB/c mice was studied. Different doses of HA DNA vaccine were used to protect BALB/c mice. 50 渭 g / g 10 渭 g / g 10 渭 g / g 5 渭 g / g 1 渭 g / g DNA vaccine with 50 渭 g / g 10 渭 g / g 10 渭 g / g DNA vaccine and 10 渭 g / g HA / na / DNA vaccine respectively. The mice were inoculated with 5 渭 g / 1 渭 g of BALB/c. Four weeks after inoculation, the mice were attacked with a lethal dose of influenza virus B / Ibaraki / 2 / 85, and the virus was attacked three days after the attack. Serum samples were taken to detect anti-HA and anti-na antibodies, and lung virus contents were determined to observe survival rate and weight loss of mice. The results showed that the vaccine was 100 渭 g HA DNA vaccine. The mice inoculated with 10 渭 g na DNA vaccine and 5 渭 g DNA vaccine were all alive. The following conclusions are drawn: one dose of DNA vaccine can provide a complete and effective protection for mice, and the combined DNA vaccine has a stronger protective effect than a single DNA vaccine. Na DNA vaccine was a little stronger than HA DNA vaccine. In order to study the effect of CpG motif as immune adjuvant. In this study, we constructed plasmid: pcD3dcontaining CpG motif (CpG)HA) and plasmid pcD3d-HA-HA without CpG motif and used different dosages. BALB/c mice were immunized with different immunization times (once or twice), four weeks after first immunization (or one week after booster immunization). Mice were attacked with a lethal dose of influenza virus B / Ibaraki / 2 / 85. Three days after the attack, serum samples were taken to detect anti-HA IgG antibodies. The survival rate and body weight of mice were observed. The results showed that the DNA vaccine containing CpG motif could further improve the antibody level. Experimental results show that CpG motif can effectively improve the ability of resisting influenza B virus attack in mice.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1

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