ANIMAL MODELS TO DESIGN AND EVALUATE IMPROVED VZV VACCINES

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 开发有效的艾滋病疫苗仍然是艾滋病毒研究的最优先事项之一。减毒的水痘-带状疱疹病毒(VZV)OKA活疫苗对预防水痘和带状疱疹安全有效，也有可能作为针对其他病原体的重组疫苗，包括人类免疫缺陷病毒(HIV)。猴水痘模型利用猴水痘病毒(SVV)，提供了一种评价重组水痘疫苗候选的方法。构建了表达猴免疫缺陷病毒(SIV)env和Gag抗原的重组SVV(RSVV)疫苗病毒。被测试的假设是，减毒的rSVV-SIV活疫苗将在恒河猴体内诱导针对SIV的免疫反应，并提供对SIV攻击的保护。结果表明，rSVV-SIV疫苗可诱导免疫猕猴产生较低水平的SIV中和抗体和细胞免疫应答，并能显著降低病原SIVmac251-CX-1静脉攻击后的病毒载量。 作为先前研究的继续，这项研究评估了额外的免疫学参数，以进一步确定这些动物的保护相关性。流式细胞仪检测CD3、CD4、CD8、CD28、CD95和Ki67抗体在刺激记忆淋巴细胞亚群中的差异。细胞内细胞因子分析检测免疫和攻击后PBMCs的功能特征。对免疫后第14天、SIV攻击当天和SIV攻击后第231天的冻存样本进行评估。标本用SIV多肽刺激，CD3、CD4、CD8表面标志及IL-2、肿瘤坏死因子-A、干扰素-g染色。结果显示，总体上，与SIV环境特异性反应相比，实验免疫动物具有更多多功能的CD4+和CD8+T细胞SIVgag特异性反应。CD4T辅助细胞中细胞增殖和抗原特异性多功能细胞因子反应的增加可能是控制疫苗接种和SIV攻击猕猴病毒载量的关键。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The development of an effective AIDS vaccine remains one of the highest priorities in HIV research. The live, attenuated varicella-zoster virus (VZV) Oka vaccine, safe and effective for prevention of chickenpox and zoster, also has potential as a recombinant vaccine against other pathogens, including human immunodeficiency virus (HIV). The simian varicella model, utilizing simian varicella virus (SVV), offers an approach to evaluate recombinant varicella vaccine candidates. Recombinant SVV (rSVV) vaccine viruses expressing simian immunodeficiency virus (SIV) env and gag antigens were constructed. The hypothesis tested was that a live, attenuated rSVV-SIV vaccine will induce immune responses against SIV in the rhesus macaques and provide protection against SIV challenge. The results demonstrated that rSVV-SIV vaccination induced low levels of neutralizing antibodies and cellular immune responses to SIV in immunized rhesus macaques and significantly reduced viral loads following intravenous challenge with pathogenic SIVmac251-CX-1. As a continuation of the previous study, this study evaluated additional immunological parameters to further define correlates of protection in these animals. Flow cytometry was conducted to show differences in stimulated memory lymphocyte subpopulations using CD3, CD4, CD8, CD28, CD95 and KI67 antibodies. Intracellular cellular cytokine assays tested functional characteristics of PBMCs following vaccination and challenge. Cryopreserved samples harvested 14 days following immunization, day of SIV challenge, and day 231 post SIV challenge were evaluated. Samples were stimulated with SIV peptides, stained with CD3, CD4, and CD8 surface markers and IL-2, TNF-A, and IFN-g. Results showed overall that experimental vaccinated animals have more polyfunctional CD4+ and CD8+ T cell SIVgag-specific responses compared with SIV env-specific responses. Increases in cellular proliferation and antigen specific polyfunctional cytokine responses in CD4 T helper cells may be crucial to control viral loads in vaccinated and SIV challenged macaques.