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有效，产生免疫反应 它可以对抗病毒、细菌和肿瘤。然而机制 支持这种方法的因素仍然不明确。这个提议是 旨在分析这些机制，并利用累积的 操纵和优化 DNA 疫苗的知识。该提案有四个 具体目标。目标 1. 鉴定摄取和表达的细胞 肌肉注射后的 DNA，将抗原呈递给 T 细胞。众所周知，DNA 在肌肉细胞中表达； APC 也是吗 转染？我们将使用克隆的CTL作为探针来识别细胞 DNA 免疫后实际上呈递抗原。目标 2. 至 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 并不意味着它们可以引起反应。我们可以使用细胞分选吗？ 转移以鉴定负责诱导免疫的细胞？ 目标 3. 评估抗原释放在 DNA 免疫中的作用，以及 以确定潜在的机制。是否诱导抗体和CD4 T细胞需要释放抗原进入体液相吗？如果是这样，什么 此版本的机制是什么？ T 细胞介导的裂解是否发挥作用 如果是的话，穿孔素和 fas 通路的作用是什么？是否 T 细胞裂解随后会限制免疫反应吗？目标 4. 使用 积累了优化 DNA 免疫的知识。知识来自 目标 1-3 将汇集在一起以优化抗体的诱导， CD4 T 细胞和 CD8 T 细胞可能各自具有独特的 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.