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Studies on in vitro expression and in vivo immunogenicity of a recombinant MVA HIV vaccine

机译:重组MVA HIV疫苗的体外表达及体内免疫原性研究

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Here we conduct dose-response studies for in vitro expression and in vivo immunogenicity for a recombinant modified vaccinia Ankara (MVA) vaccine that expresses HIV Gag, Pol, and Env proteins. The dose-response studies for in vitro expression used fluorescent-activated cell sorting to score Gag- and Env-expressing cells and showed good increases for antigen expression with increasing MVA dose. In these studies, a 1000-fold increase in the dose of MVA resulted in a 300-fold increase in the frequency of antigen-expressing cells. In contrast, dose-response studies for in vivo immunogenicity showed <10-fold increases in elicited T cells and Ab for 100-1000-fold increases in the dose of inoculated MVA. These studies used intracellular cytokine assays to enumerate responding CD8 and CD4 T cells and an ELISA to score anti-Env Ab. The shallow dose-response curves for immunogenicity were observed post priming as well as post boosting of an MVA or a DNA prime.
机译:在这里,我们进行了表达HIV Gag,Pol和Env蛋白的重组修饰牛痘安卡拉(MVA)疫苗的体外表达和体内免疫原性的剂量反应研究。体外表达的剂量反应研究使用荧光激活细胞分选法对表达Gag和Env的细胞进行评分,并显示随着MVA剂量的增加,抗原表达也有很好的提高。在这些研究中,MVA剂量增加1000倍导致抗原表达细胞频率增加300倍。相比之下,体内免疫原性的剂量反应研究表明,所诱发的T细胞增加了<10倍,Ab接种的MVA的剂量增加了100-1000倍。这些研究使用细胞内细胞因子测定法来计数响应的CD8和CD4 T细胞,并使用ELISA来对抗Env Ab进行评分。初免以及加强MVA或DNA初免后观察到了较浅的免疫原性剂量反应曲线。

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