TRANSTHYRETIN AND HIV-RELATED NEUROLOGIC DISEASE

转甲状腺素蛋白和 HIV 相关神经系统疾病

• 批准号: 2379701
• 负责人: WALTER ROYAL
• 金额: $ 10.72万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-04-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2379701
• 关键词: AIDS dementia complex; HIV infections; antiviral antibody; chemical binding; cross immunity; enzyme linked immunosorbent assay; high performance liquid chromatography; human immunodeficiency virus 1; human tissue; immunocytochemistry; in situ hybridization; laboratory rabbit; messenger RNA; polymerase chain reaction; retinoid binding proteins; thyroid hormone binding protein; thyroid hormones; virus antigen

HIV-1 p24 antigen can be detected in the CSF of seropositive individuals with dementia more frequently than in non-demented subjects. A high degree of amino acid sequence homology has been demonstrated to exist between HIV p24 and transthyretin (TTR), a molecule which binds T4, T3 and the retinol/retinol-binding protein complex (holo-RBP). The long term goals of this proposal are to determine if the structural similarity between TTR and HIV-1 p24 can be associated with antigenic cross- reactivity between TTR and HIV p24, impaired TTR function, and altered localization and synthesis of TTR in individuals with HIV-related neurologic disease. The aims of the proposed project are (1) to characterize HPLC protein fractions in CSF samples and to quantitate TTR an dp24 antigen levels in the protein fraction and in blood of seropositive individuals with and without dementia and in individuals are cross-reactive with TTR and to develop sensitive assays for detecting p24 antigens in samples that are negative for p24 antigen using currently available assays (3) to determine if specific binding occurs between TTR and HIV-1 p24 antigen and whether HIV-1 p24 antigen competes with the binding of TTE to T4 and holo-RBP; and (4) to quantitate TTR, p24 antigens, and messenger RNA (mRNA) levels for these molecules in liver and nervous system tissue lysates from seropositive individuals and seronegative controls will be separated using HPLC ion exchange chromatography followed by identification of protein species in the fractions on electrophoretic gel and immunoblots and by quantitation of protein species in the fractions on electrophoretic gels and immunoblots and by quantitation of p24 antigen and TTR in immunoassays. Antigenic cross-reactivity between p24 antigen and serum, human TTR, and recombinant HIV-1 p24 and p24 peptides. The ability of p24 antigen to inhibit TTR binding to thyroid hormone and RBP will be assayed in competitive binding studies. Tissue levels of TTR, p24 antigen, and specific mRNA will be measure in antigen capture ELISA assays and using the polymerase chain reaction assays, respectively. The morphologic localization of antigen and mRNA will be determined using immunocytochemical and in situ hybridization techniques.

在血清阳性个体的CSF中可以检测到HIV-1 p24抗原 痴呆症的频率比非痴呆受试者更频繁。 高 已证明氨基酸序列同源性的程度存在 在HIV p24和经甲状腺素素（TTR）之间，一种结合T4，T3的分子 以及视黄醇/视黄醇结合蛋白复合物（Holo-RBP）。 长 该提案的术语目标是确定结构相似性是否 TTR和HIV-1之间的P24可以与抗原交叉有关 TTR和HIV P24之间的反应性，TTR功能受损并改变 与HIV相关的个体中TTR的定位和合成 神经疾病。 拟议项目的目的是（1） 表征CSF样品中的HPLC蛋白分数，并定量TTR 蛋白质分数和血液中的DP24抗原水平 具有和没有痴呆症的血清阳性个体是 与TTR进行交叉反应，并开发用于检测P24的敏感测定法 样品中使用当前对P24抗原阴性的抗原 可用测定（3）以确定TTR之间是否发生特定结合 HIV-1 p24抗原以及HIV-1 p24抗原是否与 TTE与T4和Holo-RBP的结合； （4）定量TTR，P24 这些分子的抗原和信使RNA（mRNA）水平 以及来自血清阳性个体的神经系统组织裂解物， 血清调节控制将使用HPLC离子交换分离 色谱法随后鉴定了蛋白质物种 电泳凝胶和免疫印迹的分数以及通过定量 电泳凝胶和免疫印迹的馏分中的蛋白质物种 并通过在免疫测定中定量p24抗原和TTR。 抗原 p24抗原和血清，人类TTR和 重组HIV-1 P24和P24肽。 p24抗原的能力 抑制与甲状腺激素和RBP的TTR结合将在 竞争性结合研究。 TTR，p24抗原和组织水平 特定的mRNA将在抗原捕获ELISA分析中进行测量并使用 聚合酶链反应分别分析。 形态学 抗原和mRNA的定位将使用 免疫细胞化学和原位杂交技术。