嗜肺军团菌免疫原蛋白核酸疫苗诱导的小鼠免疫应答及其保护力论文.docVIP

　　嗜肺军团菌免疫原蛋白核酸疫苗诱导的小鼠免疫应答及其保护力论文 .freelmune response and protective efficacy in BALB/c mice induced by DNA vaccine encoding immunogenic protein of Legionella pneumophila Abstract AIM: To study the immune response in mice induced by DNA vaccine encoding immunogenic protein of Lgeionella pneumophila (LP) and to examine its protective efficacy against LP in mice. METHODS: The eukaryotic expression rebinant plasmid of LP immunogenic protein gene ( pcDNA3.1ip) munize BALB/c female mice intramuscularly, and antigen specific antibodies, lymphocyte proliferative response, IFNγ production and cytotoxic Tlymphocyte response of the immunized mice ice immunized by the pcDNA3.1ip repeatedly ber of survivors in each group ice oral and cellular immunity in the immunized mice. The results indicated that the immunogenicity of pcDNA3.1ip ice against infection after LP (10LD50) challenge. CONCLUSION: The DNA vaccine encoding immunogenic protein of LP can be used as a candidate for novel vaccine design. Keyophila; immunogenic protein; immunogenicity; DNA vaccine 摘 要 目的: 研究嗜肺军团菌免疫原蛋白核酸疫苗诱导的小鼠免疫原性以及对LP感染小鼠的保护能力。方法: 用嗜肺军团菌免疫原蛋白基因真核表达重组质粒pcDNA31ip作为DNA疫苗免疫BALB/c小鼠, 检测免疫小鼠体内抗原特异性抗体水平、 脾淋巴细胞增殖活性、 IFNγ产生水平和CTL特异杀伤活性等指标, 以评价疫苗的免疫原性。真核表达重组质粒pcDNA31ip DNA疫苗重复免疫BALB/c小鼠2次, 末次免疫2周后,.freelophila, Lp）, 有15个血清型, 其中以血清Ⅰ型最为重要。军团菌广泛存在于自然环境水和供水系统中, 人因吸入含有军团菌的气溶胶而感染, 并在肺泡上皮细胞和单核巨噬细胞内增殖。该病目前尚无有效的疫苗用于预防。军团菌的外膜蛋白与其致病密切相关。相对分子质量（Mr）为2900的外膜蛋白（29 kD immunogenic protein, IP）为1～10血清型所共有, 具有种的特异性, 是机体免疫应答反应的主要免疫原, 由嗜肺军团菌主要免疫原基因ip编码产生1。Mr2900外膜蛋白也与细菌的毒力密切相关, 具有黏附作用、 抗吞噬作用等。我们将已构建好的嗜肺军团菌ip基因真核重组质粒pcDNA31ip作为DNA疫苗免疫BALB/c小鼠, 观察其免疫原性和免疫保护性。 1 材料和方法 1.1 材料 嗜肺军团菌菌株Lp1型标准株及军团菌兔抗血清由中国疾病预防控制中心传染病控制所惠赠, 本室保存。质粒pcDNA31(+)由本室保存, 重组质粒pcDNA31ip由作者在以往的研究中构建。Omega无内毒素质粒大提取试剂盒购自天泰试剂公司, BPB Biomedicals.Inc(USA)小鼠IFNγ检测试剂盒购自豪亿试剂公司, 辣根过氧化物酶标记的羊抗兔抗体及其他试剂均购自博瑞克试剂公司。BALB/c 小鼠, 雌性, 6~8周龄, 质量18 g左右, 由四川大学华西实验动物中心提供。 1.2 方法 1.2.1 动物免疫 大量抽取无内毒素质粒pcDNA31(+)和重组质粒pcDNA31ip, 在紫外分光光度计上测量吸收值, 计算DNA含量并分析其纯度。BALB/c雌性小鼠18只, 随机分为2组, 每组9只。每只小