本文主要研究内容
作者张涵(2019)在《美洲型PRRSV二价核酸疫苗的制备及小鼠的免疫研究》一文中研究指出:猪繁殖与呼吸综合征(Procine Reproductive and Respiratory Syndrome,PRRS 最早于1987年在美国报道,自爆发以来,给养猪业造成了严重的经济损失。在1995年左右传入中国,引起该病的的病原为猪繁殖与呼吸综合征病毒(Procine Reproductive and Respiratory Syndrome Virus,PRRSV)。我国流行的主要是美洲型PRRSV及其变异株,近些年来我国PRRSV谱系呈多样化趋势,增加了我国对PRRS的预防和治疗的难度。JXA1-like谱系毒株是PRRS爆发以来比较流行的高致病性PRRSV(HP-PRRSV),而NADC30-like谱系毒株是近些年流行的HP-PRRSV。通过这两个谱系构建的二价核酸疫苗将是一种非常理想的防控PRRSV的新型疫苗。从某发病猪场采集样本采集到一株PRRSV将其命名为YN-L株。对其进行全基因扩增,然后对其Nsp2、GP5基因和全基因组进行遗传进化分析及同源性比对,并对其Nsp2、GP5基因编码的氨基酸的序列进行比对,证实该YN-L株为JX41-like谱系毒株。对YN-L毒株的理化特性进行研究,结果表明,YN-LQ毒株不耐热、不耐酸和碱,对乙醚、氯仿和紫外线敏感,对胰蛋白酶和超声波不敏感。将YN-LQ株的ORF5基因与NADC30-Lie的ORF5基因通过自裂解Linker进行连接,再与pVAX1载体连接构建二价核酸疫苗,转染至BHK细胞,用蛋白免疫印迹(Western Blot)等方法进行验证。结果显示,二价核酸疫苗可表达GP5(JXA1-Like)-Linker-GP5(NADC30-Like)目的蛋白。将所构建的二价核酸疫苗进行小鼠免疫研究,实验结果显示,二价核酸疫苗加佐剂组可提高小鼠的体液免疫和细胞免疫,具有较好的免疫原性。IL-4和IFN-γ细胞因子检测显示免疫35d时,二价核酸疫苗加佐剂组均高于二价核酸疫苗组。中和抗体检测显示免疫35d时,二价核酸疫苗加佐剂组高于其它组,中和抗体滴度最高达1:17.77。CD3~+CD4~+和CD3~+CD8~+血清细胞因子检测结果显示35d时,二价核酸疫苗加佐剂组均高于二价核酸疫苗组。T淋巴细胞增殖实验结果显示:特异性刺激组的刺激指数(SI)高于非特异性刺激组,二价核酸疫苗加佐剂组和二价核酸疫苗组高于其它组。在特异性刺激中,二价核酸苗加佐剂组高于二价核酸疫苗组;在非特异性刺激中,二价核酸疫苗加佐剂组也高于二价核酸疫苗组。综上,联合佐剂可提高二价核酸疫苗的免疫效果,为美洲型PRRSV疫苗的进一步研发奠定基础。
Abstract
zhu fan shi yu hu xi zeng ge zheng (Procine Reproductive and Respiratory Syndrome,PRRS zui zao yu 1987nian zai mei guo bao dao ,zi bao fa yi lai ,gei yang zhu ye zao cheng le yan chong de jing ji sun shi 。zai 1995nian zuo you chuan ru zhong guo ,yin qi gai bing de de bing yuan wei zhu fan shi yu hu xi zeng ge zheng bing du (Procine Reproductive and Respiratory Syndrome Virus,PRRSV)。wo guo liu hang de zhu yao shi mei zhou xing PRRSVji ji bian yi zhu ,jin xie nian lai wo guo PRRSVpu ji cheng duo yang hua qu shi ,zeng jia le wo guo dui PRRSde yu fang he zhi liao de nan du 。JXA1-likepu ji du zhu shi PRRSbao fa yi lai bi jiao liu hang de gao zhi bing xing PRRSV(HP-PRRSV),er NADC30-likepu ji du zhu shi jin xie nian liu hang de HP-PRRSV。tong guo zhe liang ge pu ji gou jian de er jia he suan yi miao jiang shi yi chong fei chang li xiang de fang kong PRRSVde xin xing yi miao 。cong mou fa bing zhu chang cai ji yang ben cai ji dao yi zhu PRRSVjiang ji ming ming wei YN-Lzhu 。dui ji jin hang quan ji yin kuo zeng ,ran hou dui ji Nsp2、GP5ji yin he quan ji yin zu jin hang wei chuan jin hua fen xi ji tong yuan xing bi dui ,bing dui ji Nsp2、GP5ji yin bian ma de an ji suan de xu lie jin hang bi dui ,zheng shi gai YN-Lzhu wei JX41-likepu ji du zhu 。dui YN-Ldu zhu de li hua te xing jin hang yan jiu ,jie guo biao ming ,YN-LQdu zhu bu nai re 、bu nai suan he jian ,dui yi mi 、lv fang he zi wai xian min gan ,dui yi dan bai mei he chao sheng bo bu min gan 。jiang YN-LQzhu de ORF5ji yin yu NADC30-Liede ORF5ji yin tong guo zi lie jie Linkerjin hang lian jie ,zai yu pVAX1zai ti lian jie gou jian er jia he suan yi miao ,zhuai ran zhi BHKxi bao ,yong dan bai mian yi yin ji (Western Blot)deng fang fa jin hang yan zheng 。jie guo xian shi ,er jia he suan yi miao ke biao da GP5(JXA1-Like)-Linker-GP5(NADC30-Like)mu de dan bai 。jiang suo gou jian de er jia he suan yi miao jin hang xiao shu mian yi yan jiu ,shi yan jie guo xian shi ,er jia he suan yi miao jia zuo ji zu ke di gao xiao shu de ti ye mian yi he xi bao mian yi ,ju you jiao hao de mian yi yuan xing 。IL-4he IFN-γxi bao yin zi jian ce xian shi mian yi 35dshi ,er jia he suan yi miao jia zuo ji zu jun gao yu er jia he suan yi miao zu 。zhong he kang ti jian ce xian shi mian yi 35dshi ,er jia he suan yi miao jia zuo ji zu gao yu ji ta zu ,zhong he kang ti di du zui gao da 1:17.77。CD3~+CD4~+he CD3~+CD8~+xie qing xi bao yin zi jian ce jie guo xian shi 35dshi ,er jia he suan yi miao jia zuo ji zu jun gao yu er jia he suan yi miao zu 。Tlin ba xi bao zeng shi shi yan jie guo xian shi :te yi xing ci ji zu de ci ji zhi shu (SI)gao yu fei te yi xing ci ji zu ,er jia he suan yi miao jia zuo ji zu he er jia he suan yi miao zu gao yu ji ta zu 。zai te yi xing ci ji zhong ,er jia he suan miao jia zuo ji zu gao yu er jia he suan yi miao zu ;zai fei te yi xing ci ji zhong ,er jia he suan yi miao jia zuo ji zu ye gao yu er jia he suan yi miao zu 。zeng shang ,lian ge zuo ji ke di gao er jia he suan yi miao de mian yi xiao guo ,wei mei zhou xing PRRSVyi miao de jin yi bu yan fa dian ding ji chu 。
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论文作者分别是来自延边大学的张涵,发表于刊物延边大学2019-07-16论文,是一篇关于美洲型论文,二价核酸疫苗论文,联合佐剂论文,免疫原性论文,延边大学2019-07-16论文的文章。本文可供学术参考使用,各位学者可以免费参考阅读下载,文章观点不代表本站观点,资料来自延边大学2019-07-16论文网站,若本站收录的文献无意侵犯了您的著作版权,请联系我们删除。
标签:美洲型论文; 二价核酸疫苗论文; 联合佐剂论文; 免疫原性论文; 延边大学2019-07-16论文;