CORE--VIROLOGY

核心--病毒学

• 批准号: 6302604
• 负责人: STEPHEN A SPECTOR
• 金额: $ 17.13万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-12-01 至 2000-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6302604
• 关键词: AIDS dementia complex; HIV infections; biomarker; biomedical facility; cerebrospinal fluid; cytomegalovirus; drug resistance; genetic strain; genotype; human immunodeficiency virus 1; human tissue; infectious encephalitis; microorganism culture; neurotropic virus; phenotype; polymerase chain reaction; psychoneuroimmunology; tissue resource /registry; virulence; virus RNA; virus genetics; virus replication; zidovudine

The objectives of the Virology Core are to elucidate the role of HIV- 1 in the development of central nervous system (CNS) impairment, to identify HIV-1-specific determinants predictive of CNS dysfunction, and to determine the importance of human cytomegalovirus (CMV) on the development of HIV-1-related CNS disease. The findings of the Virology Core will be integrated in the HNRC data base, which will enable correlations with findings of the other research cores and specific projects. Additionally, cerebrospinal fluid (CSF), peripheral blood mononuclear cells and plasma specimens, viral isolates, and nucleic acids (including PCR amplified products) will be stored in the HNRC specimen bank and made available to other investigators for further virologic investigations. The specific aims of the Virology Core are: (1) To identify virologic markers and kinetics of HIV-1 replication which are predictive of HIV-1-related CNS disease. We hypothesize that HIV-1 load as determined by quantitative RNA, and decreased antiretroviral sensitivity as determined by phenotypic and genotypic analyses of HIV-1 isolates and nucleic acid from CSF will correlate with the presence and development of neurocognitive impairment. Additionally, we will examine the kinetics of HIV-1 replication within the CSF, and correlate these findings with replication kinetics in plasma and the presence of neurocognitive impairment: (2) To determine the pathogenetic significance of enhanced HIV-1 replication in macrophages cocultured with endothelial cells in the development of CNS impairment. The hypothesis to be tested is that HIV-1 strains with the greatest enhanced replication in macrophage-endothelial cell cultures will be most likely to enter the CNS and be correlated with the development of neurocognitive impairment. Additionally, we propose to develop strategies to block macrophage/binding to brain microvascular endothelial cells such that HIV-1 replication in the brain will be decreased, and thus, reduce the incidence of HIV-1-related CNS impairment; (3) To determine the role of CMV in the development of CNS disease in HIV-1-infected persons and to define the importance of antiviral resistance in persons who develop CMV- related CNS disease. We hypothesize that as HIV-1-infected persons survive for prolonged periods with low CD4+ lymphocyte counts that viruses other than HIV-1, particularly CMV, will become increasingly important copathogens contributing to the development of neurocognitive impairment. Additionally, we hypothesize that as oral ganciclovir prophylaxis is used to prevent CMV disease that resistant strains of CMV will become an important cause of CNS disease. For these studies, we will perform qualitative and quantitative PCR assays to monitor patients participating in the proposed CMV study. Additionally, clinical isolates and DNA obtained from the CSF of these study participants will be evaluated for phenotypic and genotypic resistance to antivirals (ganciclovir, foscamet and HPMPC) most often used to treat HIV-1-infected persons With CMV disease. The findings of the Virology Core will improve the understanding of HIV-1 pathogenesis and enable the design of treatment studies directed toward the prevention of CNS dysfunction in persons identified to be at highest risk for developing abnormalities.

病毒学核心的目标是阐明 HIV-1 在 中枢神经系统（CNS）损伤的发展，以确定 HIV-1特异性决定因素可预测中枢神经系统功能障碍，并 确定人类巨细胞病毒 (CMV) 对发育的重要性 HIV-1相关的中枢神经系统疾病。病毒学核心的研究结果将是 集成到 HNRC 数据库中，这将能够与 其他研究核心和具体项目的发现。此外， 脑脊液 (CSF)、外周血单核细胞和血浆 样本、病毒分离物和核酸（包括 PCR 扩增 产品）将存储在 HNRC 样本库中并可供 其他研究人员进行进一步的病毒学研究。具体的 病毒学核心的目标是： (1) 识别病毒学标记并 HIV-1复制动力学可预测HIV-1相关中枢神经系统 疾病。我们假设 HIV-1 载量由定量 RNA 确定， 并根据表型和抗逆转录病毒敏感性降低 HIV-1 分离株和脑脊液核酸的基因型分析将 与神经认知障碍的存在和发展相关。 此外，我们将检查 HIV-1 复制的动力学 CSF，并将这些发现与血浆中的复制动力学相关联 是否存在神经认知障碍： (2) 确定 巨噬细胞中 HIV-1 复制增强的发病意义 在中枢神经系统损伤的发展中与内皮细胞共培养。 要检验的假设是 HIV-1 毒株具有最大的 巨噬细胞-内皮细胞培养物中复制的增强将是最重要的 可能进入中枢神经系统并与发育相关 神经认知障碍。此外，我们建议制定战略 阻止巨噬细胞/与脑微血管内皮细胞结合，例如 HIV-1在大脑中的复制将会减少，从而减少 HIV-1 相关中枢神经系统损伤的发生率； (3)确定角色 CMV 在 HIV-1 感染者中枢神经系统疾病发展中的作用 定义抗病毒耐药性对于 CMV 感染者的重要性 相关中枢神经系统疾病。我们假设 HIV-1 感染者能够存活下来 长时间处于低 CD4+ 淋巴细胞计数状态 比 HIV-1，特别是 CMV，将变得越来越重要 导致神经认知障碍发展的共同病原体。 此外，我们假设使用口服更昔洛韦进行预防 为了预防 CMV 疾病，CMV 耐药株将成为 中枢神经系统疾病的重要病因。对于这些研究，我们将执行 定性和定量 PCR 检测来监测参与的患者 在拟议的 CMV 研究中。此外，临床分离株和 DNA 从这些研究参与者的 CSF 中获得的信息将被评估 对抗病毒药物（更昔洛韦、福卡美）的表型和基因型耐药性 和 HPMPC）最常用于治疗感染 CMV 的 HIV-1 感染者 疾病。病毒学核心的发现将增进对病毒学的理解 HIV-1 发病机制并能够设计针对性的治疗研究 旨在预防被确定患有中枢神经系统功能障碍的人 发生异常的风险最高。