ENHANCEMENT OF DNA VACCINE IMMUNOGENICITY

DNA免疫是一种潜在的重要疫苗接种方法。它 在许多动物模型中显示有效， 可以对抗病毒细菌和肿瘤然而，机制 这一方法的基础仍然没有明确界定。这项建议是 旨在分析这些机制，并使用累积的 操纵和优化DNA疫苗的知识。该提案有四个 具体目标。目标1。来识别哪些细胞摄取并表达 肌肉注射后，DNA，并提出抗原T 细胞已知DNA在肌肉细胞中表达; APC也 被感染了我们将使用克隆的CTL作为探针来鉴定细胞 在DNA免疫后呈递抗原。目标二。到 精确识别哪些细胞诱导免疫，并确定是否 肌肉细胞很重要。T细胞可以识别细胞， 这并不意味着它们可以引起反应。我们能不能用细胞分类和 转移来识别负责诱导免疫的细胞？ 目标3。评价抗原释放在DNA免疫中的作用， 来识别潜在的机制。诱导抗体和CD 4 + T细胞需要抗原释放进入体液相？如果是的话， 这种释放的机制是什么T细胞介导的溶解作用是否 作用，如果是这样，穿孔素和Fas通路的作用是什么？并 T细胞溶解随后限制免疫反应？目标4。使用 积累了优化DNA免疫的知识。上的知识 目标1-3将被集中在一起以优化抗体的诱导， CD 4 + T细胞和CD 8 + T细胞中的每一种可能都具有独特的 最佳反应的要求。

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.