H5、H7亚型流感多表位核酸疫苗的分子设计、构建及实验免疫研究

发布时间：2018-06-16 19:44

本文选题：流感 + 多表位　；参考：《吉林大学》2010年博士论文

【摘要】： 流感是一种由流感病毒引发的传染病,患者以发热、粘膜充血、急性呼吸道症状为主。该病发病率高,破坏性强,从二十世纪三十年代人类发明流感疫苗至今,每年仍有25～50万人死于流感。季节性流感以H1、H3亚型为主,而禽流感H5、H7亚型也不断出现人类感染病例。流感病毒不断变异,随时都有可能进化成为大流行流感毒株,世界上很多国家都在研发新型流感疫苗以应对多种亚型流感大流行。本研究利用生物信息学方法、免疫学知识,对H1、H3亚型流感病毒的T、B细胞表位进行预测与筛选,获得CTL表位3条:H1HA545-553、H3HA546-554、H3NP189-197,B和Th细胞表位4条:H1HA142~156、H1HA212~226、H3HA247~261、H3HA287~301。结合已发表的表位,设计、合成CTL、B/Th多表位表达盒。以H5HA、H7HA、H1NP、多表位表达盒为主要目的基因,pVAX1为载体构建用来预防H5、H7、H1、H3亚型流感的多表位核酸疫苗。将所构建的疫苗载体转染BHK细胞,通过RT-PCR和IFA验证核酸疫苗抗原成分可以在真核细胞中成功表达。利用小鼠作为哺乳动物模型,对所构建的复合多表位核酸疫苗免疫原性进行研究,表明构建的核酸疫苗在诱导细胞免疫及体液免疫方面具有显著优势。本实验的创新之处在于:1)依靠H5、H7亚型流感主要抗原成分,结合H1、H3抗原的功能表位构建了一种新型复合多表位核酸疫苗,目的是通过疫苗接种来预防H5、H7、H1、H3多种亚型流感病毒;2)预测多条H1、H3亚型流感功能表位,并将表位设计成能够发挥功能的多表位表达盒;3)通过小鼠实验免疫评价构建核酸疫苗的免疫原性。
[Abstract]:Influenza is an infectious disease caused by influenza virus, with fever, mucosal congestion and acute respiratory symptoms. The incidence of the disease is high and destructive. Since the invention of influenza vaccine in 1930s, 25 ~ 500000 people still die from influenza every year. Seasonal influenza mainly consists of H _ 1 H _ 3 subtype, and avian influenza H _ 5 H _ 5 H _ 7 subtype also presents human infection cases. Influenza viruses are mutating and could evolve into pandemic strains at any time. Many countries around the world are developing new influenza vaccines to cope with multiple subtypes of influenza pandemics. In this study, using bioinformatics and immunological knowledge, the T cell epitopes of H1H3 subtype influenza virus were predicted and screened. Three CTL epitopes: H3HA545-553H3HA546-554H3NP189-1979B and Th cell epitopes were obtained. Combined with the published epitopes, CTL B / T h multiepitope expression cassette was designed and synthesized. The multiepitope nucleic acid vaccine against H5, H7, H1, H1 and H3 subtype influenza was constructed by using H5 H7 H7 HAH1 NP1 and multiepitope expression cassette as main target gene pVAX1 as vector for the prevention of H5, H7, H1, H1, H1, H1, H3 subtype influenza. The constructed vaccine vector was transfected into BHK cells. RT-PCR and IFA were used to verify that the antigen components of nucleic acid vaccine could be successfully expressed in eukaryotic cells. The immunogenicity of the complex polyepitope nucleic acid vaccine was studied by using mouse as a mammalian model. The results showed that the constructed nucleic acid vaccine had significant advantages in inducing cellular and humoral immunity. The innovation of this experiment is that a novel compound polyepitope nucleic acid vaccine was constructed based on the main antigen components of H5 and H7 subtype influenza and the functional epitopes of H1H3 antigen. The aim of this study was to predict multiple functional epitopes of H1 / H3 subtype influenza virus by vaccinating against H5 / H7 / H1 / H1 / H3 subtype influenza viruses. The immunogenicity of nucleic acid vaccine was evaluated by mouse experimental immunization.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R392

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