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首页> 外文学位 >Characterization of viral envelopes and cytokine responses among rhesus macaques immunized with vesicular stomatitis virus-based simian immunodeficiency virus (SIV) vaccines and challenged with pathogenic SIV.
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Characterization of viral envelopes and cytokine responses among rhesus macaques immunized with vesicular stomatitis virus-based simian immunodeficiency virus (SIV) vaccines and challenged with pathogenic SIV.

机译：基于水泡性口炎病毒的猿猴免疫缺陷病毒（SIV）疫苗免疫并受到病原性SIV攻击的恒河猴的病毒包膜和细胞因子应答的特征。

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摘要

Precise identification of the transmitted/founder virus envelope sequences responsible for infection is critical for understanding the viral transmission and designing effective vaccines. Single Genome Amplification (SGA) was utilized to identify unambiguously the transmitted viruses. In the SGA technique, viral RNA is isolated from plasma; cDNA is made and amplified by nested PCR. The amplicons showing <30% of the total number of reactions (representing single virus particle as per Poisson distribution) were sequenced and further analyzed using phylogenetic tools. In the current study, vaccinated and immunized macaques were challenged intra-rectally (IR) with 4000 TCID50 of SIVsmE660. The macaques were primed and boosted with VSV (vesicular stomatitis virus) expressing SIVsmE660 Gag, Env proteins (Group I), VSV expressing SIVsmE660 Gag, Env proteins and GM-CSF (Group II) or with VSV expressing an influenza HA (hemagglutinin) protein (Group III) as the control group. After priming with the VSV vaccine vectors, the macaques were boosted with VSV/SFV (Semiliki Forest virus) replicon particles expressing SIVsm E660 Env and Gag proteins. Four of the six animals in the VSV-Gag-Env immunized group were protected from infection while 2 had a transient viral load. Only1/6 was protected from infection among VSV-GM-CSF-Gag-Env immunized animals suggesting that GM-CSF abrogated protection. All animals from the irrelevant antigen vaccinated group showed high viral loads. We first characterized the challenge SIVsmE660 stock virus. In total, 58 full-length Env clones were analyzed and the diversity of the stock was calculated as 1.4%. We obtained full-length envelope sequences from each animal that belongs to Group I (2/2) Group II (4/6) and Group III (6/6). Our preliminary data suggests that few founder viruses (1 founder lineage) were found in the protected animals from Group I as compared to 1 or more founder lineages in Group II / III animals. We also analyzed the cytokines from the plasma of the above vaccinated and challenged macaques pre and post-challenge using non-human primate 23-plex cytokine assay kit. Among the animals that were infected and had high viral loads exhibited elevated levels of IFN-gamma, TNF-alpha, MCP-1 and IL-18.
机译：准确识别负责感染的传播/基础病毒包膜序列对于理解病毒传播和设计有效疫苗至关重要。单基因组扩增（SGA）被用来明确地确定传播的病毒。在SGA技术中，病毒RNA是从血浆中分离出来的。通过巢式PCR制备并扩增cDNA。对显示<30％反应总数的扩增子（按照泊松分布表示单个病毒颗粒）进行测序，并使用系统发育工具进行进一步分析。在当前的研究中，接种和免疫的猕猴接受4000 TCID50的SIVsmE660直肠内（IR）攻击。用表达SIVsmE660 Gag的VSV（水泡性口炎病毒），Env蛋白（I组），表达SIVsmE660 Gag的VSV，Env蛋白和GM-CSF（组II）或用表达流感HA（血凝素）蛋白的VSV引发和加强猕猴。 （第三组）为对照组。用VSV疫苗载体启动后，猕猴用表达SIVsm E660 Env和Gag蛋白的VSV / SFV（Semiliki Forest病毒）复制子颗粒加强免疫。 VSV-Gag-Env免疫组的六只动物中有四只受到了保护，免受感染，而其中两只具有短暂的病毒载量。在受VSV-GM-CSF-Gag-Env免疫的动物中，只有1/6受到保护免于感染。这表明GM-CSF取消了保护。来自无关抗原疫苗接种组的所有动物均显示高病毒载量。我们首先表征了挑战SIVsmE660股票病毒。总共分析了58个全长Env克隆，种群的多样性被计算为1.4％。我们从属于组I（2/2）组II（4/6）和组III（6/6）的每只动物中获得了全长包膜序列。我们的初步数据表明，与II / III组动物中的1个或多个Founder世系相比，在I组的受保护动物中几乎没有发现Founder病毒（1个Founder世系）。我们还使用非人类灵长类动物23-plex细胞因子检测试剂盒分析了攻击前和攻击后上述免疫和攻击过的猕猴血浆中的细胞因子。在被感染并具有高病毒载量的动物中，IFN-γ，TNF-α，MCP-1和IL-18的水平升高。

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  • 作者

    Gambhira, Ratish.;

  • 作者单位

    Tulane University.;

  • 授予单位 Tulane University.;
  • 学科 Biology Microbiology.;Biology Virology.
  • 学位 Ph.D.
  • 年度 2012
  • 页码 176 p.
  • 总页数 176
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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