Functional Analysis of NSV-based HIV vectors

描述（由申请人提供）：负链RNA病毒（NSV）载体已显示出作为多种传染病疫苗的前景。虽然它们具有相似的生命周期，但NSV载体通过非常不同的机制与先天免疫（如I型干扰素反应）相互作用并调节。VSV是一种高度细胞病变病毒，通过其M蛋白关闭细胞基因表达。狂犬病毒（RV）是一种非细胞病变病毒，通过其P蛋白下调STAT功能。NDV是一种禽副粘病毒，在哺乳动物细胞中似乎没有下调先天免疫的特定机制。所有这些病毒在体内都有不同程度的复制，并在树突状细胞中引发不同的宿主信号程序。由于尚不清楚什么构成艾滋病毒疫苗的理想载体，我们现在将比较这些病毒并提取有关它们的特定特征如何调节免疫原性的信息。具有相似主干和复制周期但具有非常不同宿主-病毒相互作用的载体诱导的免疫原性参数尚未进行全面分析。该应用的总体假设是，NSV载体可以通过改变其与先天免疫系统的相互作用而得到改善。我们假设有效的HIV-1疫苗必须激活树突状细胞（dc），树突状细胞是先天免疫和适应性免疫之间的重要联系。我们将首先研究这些表达HIV-1抗原的载体对体外dc介导的免疫的影响，并确定有望在体内诱导有效适应性免疫的参数，如有效的抗原呈递和激活。这些初步研究将使我们能够提高载体的效率，并使我们对诱导hiv -1特异性免疫反应的最佳载体的构成有更好的基本了解。一旦这些参数确定，我们将在小鼠中研究异源启动-增强方法的致病性和免疫原性载体。在小鼠身上发现的最有希望的方法将在恒河猴身上进一步分析。

DESCRIPTION (provided by applicant): Negative-stranded RNA virus (NSV) vectors have shown promise as vaccines against a variety of infectious diseases. While they have similar life-cycles, NSV vectors interact with and modulate innate immunity, such as type I interferon responses, by very different mechanisms. VSV is a highly cytopathic virus that shuts down cellular gene expression by virtue of its M protein. Rabies virus (RV) is a noncytopathic virus that down-modulates STAT function by virtue of its P protein. NDV is an avian paramyxoviruses that appears not to have specific mechanisms of down-modulating innate immunity in mammalian cells. All of these viruses replicate to different extents in vivo and elicit different host signaling programs in dendritic cells. Since it is unclear what constitutes an ideal vector for HIV vaccines, we will now compare these viruses and extract information on how their specific characteristics modulate immunogenicity. A comprehensive analysis of immunogenicity parameters induced by vectors with a similar backbone and replication cycle but with very diverse set of host-virus interactions has not been performed. The overall hypothesis for this application is that NSV vectors can be improved by altering their interactions with the innate immune system. We hypothesize that an effective HIV-1 vaccine must activate dendritic cells (DCs), which are an important link between innate and adaptive immunity. We will first study the impact of these vectors expressing HIV-1 antigens on DC-mediated immunity in-vitro, and identify parameters that would be expected to induce effective adaptive immunity in-vivo, such as efficient antigen presentation and activation. These initial studies will allow us improve the efficiency of our vectors, and provide us with a better basic understanding of what constitutes an optimal vector for the induction of HIV-1-specific immune responses. Once these parameters are determined, we will study the vectors for pathogenicity and immunogenicity in heterologous prime-boost approaches in mice. The most promising approaches identified in mice will be further analyzed in rhesus macaques.