PANCREATITIS INDUCED BY COXSACKIE VIRUS B4

该实验室的长期目标是确定病毒如何 引发疾病。选择柯萨奇病毒B4(CB4)进行这些研究 因为这种病毒已经牵连到人类疾病，如1型 胰岛素依赖型糖尿病和心肌炎。使用鼠标模型系统 对于CB4诱导的胰腺炎，这项建议解决了两个主要问题。 前两个具体目标是直截了当的，重点是 致病性的分子基础： 1.决定毒力表型的病毒基因(S)的定位 利用重组、嵌合病毒产生的一种新的 无毒(CB4-P)和强毒亲本病毒(CB4-V)； 2.确定毒力的核苷酸序列的鉴定 通过对合适的cdna克隆进行DNA序列分析来进行表型分析。自.以来 可能存在多个突变，将通过以下方式分析单个基因座 将单、双等突变切除回无毒， 亲代病毒克隆。 第三个具体目标是开始探索更复杂的问题，即如何 特定突变通过检查病毒是否有毒力来决定毒力 功能和免疫原性改变。 3A。CB4-V和CB4-P是否感染胰腺内不同类型的细胞？ 这个问题将通过使用原位杂交来检测 感染组织中的病毒RNA。 3B。这些改变的基因座编码抗原决定簇吗？适当的 将对多肽进行测试，以确定它们是否能激发细胞毒性T细胞 在受感染的小鼠中的反应或抗体反应。 第四个具体目标是作为第三个具体目标的替代方案提出的。 4.通过A.作图来表征CTL对病毒感染的反应 CB4-P和CB4-V、B的CTL识别决定因素 两种病毒对CB4-V的CTL应答是否有差异 不同H-2单倍型的小鼠，C.确定组织损伤 在CB4-V感染的小鼠中观察到的是免疫介导的。 这些研究应该会在分子发病机制上产生新的数据 柯萨奇病毒感染与感染的关系 毒力和病毒功能以及毒力和免疫原性之间的关系。

The long-term objective of this laboratory is to determine how viruses cause disease. Coxsackievirus B4 (CB4) was chosen for these studies since this virus has been implicated in human diseases such as type 1 insulin-dependent diabetes and myocarditis. Using a mouse model system of CB4-induced pancreatitis, this proposal addresses two main issues. The first two specific aims are straightforward and focus on the molecular basis of pathogenicity: 1. Mapping of the viral gene(s) that determine the virulent phenotype using recombinant, chimeric viruses generated from cDNA clones of a non-virulent (CB4-P) and a highly virulent parental virus (CB4-V); 2. Identification of the nucleotide sequences that determine the virulent phenotype by DNA sequence analysis of appropriate cDNA clones. Since multiple mutations are likely, individual loci will be analyzed by resection of single, double, etc. mutations back into the avirulent, parental virus clone. The third specific aim begins to explore the more complex question of how specific mutations determine virulence by examining whether viral function and immunogenicity are altered. 3A. Do CB4-V and CB4-P infect different cell types within the pancreas? This question will be addressed by using in situ hybridization to detect viral RNA in infected tissue. 3B. Do the altered loci encode antigenic determinants? The appropriate peptides will be tested to determine if they can evoke a cytotoxic T cell response or an antibody response in infected mice. The fourth specific aim is proposed as an alternative to specific aim #3. 4. Characterization of the CTL response to viral infection by: A. Mapping of the CTL recognition determinants of CB4-P and CB4-V, B. Determining whether there is a difference in the CTL response against CB4-V between mice of different H-2 haplotypes, C. Determining if the tissue damage observed in CB4-V-infected mice is immune mediated. These studies should generate new data on the molecular pathogenesis of coxsackievirus infections in addition to information on the relationship between virulence and viral function and virulence and immunogenicity.