Coronavirus-based multigene HIV vaccine vectors

基于冠状病毒的多基因艾滋病毒疫苗载体

基本信息

批准号：
7140580
负责人：
Volker Thiel
金额：
$ 12.21万
依托单位：
CANTONAL HOSPITAL ST GALLEN
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-05 至 2007-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Coronavirus-based vectors are a promising system to genetically deliver multiple heterologous genes to specific target cells. First, coronaviruses are positive-stranded RNA viruses replicating in the cytoplasm without a DNA intermediary, making insertion of viral sequences into the host cell genome unlikely. Second, coronaviruses have the largest RNA genome known so far and a cloning capacity of more than 6 kb is expected. Third, coronaviruses display a unique transcription process resulting in the synthesis of 6-8 subgenomic mRNAs, encoding mainly the structural genes which can be replaced by multiple genes-of-interest. Fourth, the receptor of human coronavirus HCoV 229E is expressed on dendritic cells. Fifth, the natural route of coronavirus infection is the mucosal route. The long term objective of our study is to generate a novel HIV vaccine vector that (i) targets and activates DC (ii) displays sufficient cloning capacity for insertion of several HIV antigens and immunostimulatory cytokines, (iii) encodes for antigenic determinants that induce broadly neutralizing antibody responses, and (iv) can be applied via mucosal surfaces. The specific aim of this application is to provide the proof-of-principle that coronavirus vectors can induce protective antiviral humoral and cellular immune responses in vivo. To this end, we will generate mouse hepatitis virus (MHV) vectors carrying various antigens (fusion protein of the LCMV GP33 epitope KAVYNFATC and green fluorescent protein, vesicular stomatitis virus glycoprotein (VSV-G), and HIV-p24) and immunostimulatory cytokine GM-CSF. Mice will be immunized with recombinant MHV virus-like particles via different routes and the induction of antiviral CTL and Th cell responses and the production of neutralizing antibodies will be tested using well-established read-out systems. Since MHV infects mucosal tissues and is able to infect DC, recombinant MHV vectors in the context of a murine model can serve as a paradigm for the development and evaluation of coronavirus vaccine vectors in humans.

描述（由申请人提供）：基于冠状病毒的载体是一种很有前途的系统，可以将多个异源基因遗传地传递到特定的靶细胞。首先，冠状病毒是正链RNA病毒，在细胞质中复制，没有DNA中介，因此不可能将病毒序列插入宿主细胞基因组中。其次，冠状病毒拥有迄今为止已知最大的RNA基因组，预计克隆能力将超过6 kb。第三，冠状病毒表现出独特的转录过程，导致 6-8 个亚基因组 mRNA 的合成，主要编码可被多个感兴趣基因替换的结构基因。第四，人类冠状病毒HCoV 229E的受体在树突状细胞上表达。第五，冠状病毒感染的自然途径是粘膜途径。我们研究的长期目标是产生一种新型 HIV 疫苗载体，该载体（i）靶向并激活 DC（ii）表现出足够的克隆能力以插入多种 HIV 抗原和免疫刺激细胞因子，（iii）编码广泛诱导的抗原决定簇中和抗体反应，并且（iv）可以通过粘膜表面施用。该应用的具体目的是提供冠状病毒载体可以在体内诱导保护性抗病毒体液和细胞免疫反应的原理证明。为此，我们将生成携带各种抗原（LCMV GP33表位KAVYNFATC和绿色荧光蛋白的融合蛋白、水泡性口炎病毒糖蛋白（VSV-G）和HIV-p24）和免疫刺激细胞因子GM的小鼠肝炎病毒（MHV）载体-脑脊液。将通过不同途径用重组 MHV 病毒样颗粒对小鼠进行免疫，并使用完善的读出系统测试抗病毒 CTL 和 Th 细胞反应的诱导以及中和抗体的产生。由于 MHV 感染粘膜组织并能够感染 DC，因此小鼠模型中的重组 MHV 载体可以作为人类冠状病毒疫苗载体开发和评估的范例。