ENHANCEMENT OF DNA VACCINE IMMUNOGENICITY

DNA免疫是一种潜在的重要疫苗接种方法。它 已被证明在许多动物模型中起作用，产生免疫反应 它可以对抗病毒、细菌和肿瘤。然而，这些机制 支撑这一方法的因素仍然定义不清。这项建议是 旨在分析这些机制，并利用应计 掌握操纵和优化DNA疫苗的知识。该提案有四个方面 明确的目标。目的1.鉴定哪些细胞能吸收和表达 肌肉注射后的DNA，以及哪些DNA呈递T细胞抗原 细胞。已知DNA在肌肉细胞中表达；APC是否也 被改造了？我们将使用克隆的CTL作为探针来鉴定细胞 实际上是在DNA免疫后呈递抗原。目标2.目标 准确识别哪些细胞诱导免疫，并确定 肌肉细胞很重要。细胞可以被T细胞识别 这并不意味着它们可以引发反应。我们是否可以使用单元格排序& 转移以识别负责诱导免疫的细胞？ 目的3.评价抗原释放在DNA免疫中的作用 以确定潜在的机制。抗体和CD4的诱导 T细胞需要将抗原释放到体液相？如果是这样，会发生什么？ 这个版本背后的机制是什么？T细胞介导的裂解作用是否发挥作用 作用，如果是，穿孔素和Fas通路的作用是什么？会吗？ T细胞裂解会限制免疫反应吗？目标4.使用 积累了优化DNA免疫的知识。来自中国的知识 目标1-3将被结合在一起以优化抗体的诱导， CD4T细胞和CD8T细胞中的每一个都可能具有独特的 对最佳响应的要求。

DNA immunization is a potentially important approach to vaccination. It has been shown to work in many animal models, producing immune responses which can counter viruses, bacteria, and tumors. However the mechanisms which underpin this approach remain poorly defined. This proposal is aimed towards analyzing these mechanisms, and using the accrued knowledge to manipulate and optimize DNA vaccines. The proposal has four specific aims. Aim 1. To identify the cells which take up and express DNA following intramuscular injection, and which present antigen to T cells. It is known that DNA is expressed in muscle cells; are APCs also transfected? We shall use cloned CTL as probes to identify the cells actually presenting antigen following DNA immunization. Aim 2. To precisely identify which cells induce immunity, and to determine whether muscle cells are important. That cells can be recognized by T cells does not imply that they can induce responses. Can we use cell sorting & transfer to identify the cells responsible for induction of immunity? Aim 3. To evaluate the role of antigen release in DNA immunization, and to identify the underlying mechanisms. Does induction of antibody & CD4+ T cells require antigen release into the humoral phase? If so, what mechanisms underlie this release? Does T cell mediated lysis play a role, and if so, what are the roles of the perforin & fas pathways? Does T cell lysis subsequently limit the immune response? Aim 4. To use the accumulated knowledge to optimize DNA immunization. The knowledge from aims 1-3 will be drawn together to optimize induction of antibodies, CD4+ T cells, and CD8+ T cells each of which probably will have unique requirements for optimal responses.