Retroviral Antisense Genes as Future DNA Vaccines.

描述：（由申请人提供）我们发现了一种新的基因， 在HIV-1疾病进展中发挥作用。这一点是通过删除或 许多HIV-1感染者长期存活者（LTS）中的突变 多年未出现艾滋病症状， 一种减毒病毒的证据我们建议在这笔拨款中， 鉴定该基因编码的蛋白质（HAP），以开发DNA载体 能够有效表达蛋白质（HAP-DNA），并靶向HAP-DNA 递送至树突状细胞（DC）。我们还将检查猿类免疫缺陷 病毒（SIV）序列作为类似基因的证据，因为SIV 恒河猴的感染代表了AIDS发病机理的模型。我们 进一步确定HAP的免疫原性。为了确定这一点，我们计划 研究外周血淋巴细胞（PBL）或纯化的T细胞，从 HIV-1感染或非感染个体，在体外刺激后， 用HAP-DNA或对照DNA脉冲的自体DC。 增殖以及活化状态和细胞因子的定量 T细胞亚群的特征（IFN-g、IL-4、TNF-a）将在以下测定： 通过HAP-DNA脉冲的自体树突状细胞（DC）的HAP肽呈递 使用流式细胞术和酶联免疫斑点（ELISPOT）测定， IFN-g释放研究以确定DC分化的状态， 促进诱导细胞免疫和TH 1极化，将是 探索作为扩大体外初级免疫应答的机制， HAP蛋白。这些研究将有助于确定 HAP-DNA作为未来HIV-1 DNA疫苗候选者

DESCRIPTION: (Provided by Applicant) We have discovered a novel gene that may play a role in HIV-1 disease progression. This is suggested by its deletion or mutation in many long-term survivors (LTS), who are HIV-1 infected individuals who have not progressed to symptoms of AIDS for many years and who have evidence of an attenuated virus. We propose in this grant to further characterize the protein (HAP) encoded in this gene, to develop a DNA vector capable of efficiently expressing the protein (HAP-DNA), and targeting HAP-DNA delivery to dendritic cells (DCs). We will also examine simian immunodeficiency virus (SIV) sequences for evidence of an analogous gene, inasmuch as SIV infection of rhesus macaques represents a model for AIDS pathogenesis. We further want to determine HAP's immunogenicity. To determine this, we plan to study peripheral blood lymphocytes (PBL) or purified T cells, obtained from HIV-1 infected or non-infected individuals, following stimulation in vitro with autologous DCs pulsed with HAP-DNA or control-DNA(s). Quantification of proliferation, as well as the activation state and cytokine profile (IFN-g, IL-4, TNF-a), of T cell subsets, will be determined following HAP peptide presentation by HAP-DNA pulsed, autologous dendritic cells (DCs) using both flow cytometry, and an enzyme-linked immunospot (ELISPOT) assay for IFN-g release. Studies to determine the state of DC differentiation that will facilitate induction of cellular immunity and TH1 polarization, will be explored as a mechanism of expanding ex vivo primary immune responses to the HAP protein. These studies will help determine the potential and viability of HAP-DNA as a future HIV-1 DNA vaccine candidate.