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B型流感病毒核酸疫苗和CpG基序免疫增強作用研究

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  本文關(guān)鍵詞:B型流感病毒核酸疫苗和CpG基序免疫增強作用研究 出處:《湖南師范大學》2005年碩士論文 論文類型:學位論文


  更多相關(guān)文章: 流感病毒 DNA疫苗 CpG基序 血凝素 神經(jīng)氨酸酶


【摘要】:本論文介紹了B型流感病毒DNA疫苗以及CpG基序作為佐劑對其免疫增強效應的研究。 血凝素(Hemagglutinin,HA)和神經(jīng)氨酸酶(neuraminidase,NA)是B型流感病毒兩種最重要的抗原蛋白,本文首先探討了一次接種B型流感病毒(B/Ibaraki/2/85)DNA疫苗對BALB/c小鼠的免疫保護作用。分別采用不同劑量的HA DNA疫苗(100μg、50μg、10μg、5μg、1μg),NA DNA疫苗(50μg、10μg、5μg、1μg),以及HA和NA聯(lián)合DNA疫苗(10μg、5μg、1μg),一次接種BALB/c小鼠。接種后四周,用致死量流感病毒(B/Ibaraki/2/85)攻擊小鼠,病毒攻擊后三天,取血清檢測抗-HA和抗-NA抗體。同時測定小鼠肺部病毒含量,觀察記錄小鼠存活率和體重丟失情況。結(jié)果發(fā)現(xiàn):100μg HA DNA疫苗,10μg NA DNA疫苗以及5μg聯(lián)合DNA疫苗(HA+NA)接種的小鼠全部存活。結(jié)合小鼠肺部病毒含量和體重變化數(shù)據(jù),得出以下結(jié)論:一次接種DNA疫苗可以為小鼠提供完全有效的保護,聯(lián)合DNA疫苗免疫保護作用強于單一DNA疫苗,NA DNA疫苗免疫保護作用略強于HA DNA疫苗。 為了探討CpG基序作為免疫佐劑的效應,本研究構(gòu)建了包含CpG基序的質(zhì)粒:pcD3d(+CpG)HA和不包含CpG基序的質(zhì)粒pcD3d(-)HA。并采用不同的劑量,不同的免疫次數(shù)(一次或兩次),分別免疫BALB/c小鼠,初免后四周(或加強免疫后一周),用致死量流感病毒(B/Ibaraki/2/85)攻擊小鼠,病毒攻擊后三天,取血清檢測抗-HA IgG抗體,并觀察小鼠存活率和體重變化情況。結(jié)果顯示,含CpG基序的DNA疫苗能夠進一步提高抗體水平,改善小鼠病毒攻擊后的臨床體癥。實驗表明CpG基序能有效提高小鼠抗B型流感病毒攻擊的能力。
[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.
【學位授予單位】:湖南師范大學
【學位級別】:碩士
【學位授予年份】:2005
【分類號】:R392.1

【參考文獻】

相關(guān)期刊論文 前2條

1 戚鳳春,盛軍;流感疫苗的研究進展[J];中國生物制品學雜志;2004年03期

2 張燁,溫樂英,王敏,郭俊峰,李梓,郭元吉;2001年中國新分離維多利亞系乙型流感病毒的抗原性及基因特性分析[J];中華實驗和臨床病毒學雜志;2003年01期



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