Replication and Pathogenic Potential of XMRV in Humans

XMRV 在人类中的复制和致病潜力

• 批准号: 8349482
• 负责人: VINAY K. PATHAK
• 金额: $ 26.75万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8349482
• 关键词: Biological Assay; Cell Line; Cells; Chronic Fatigue Syndrome; Disease; Enzyme-Linked Immunosorbent Assay; Gammaretrovirus; HIV-1; Human; Incidence; Individual; Infection; Integrase; Integrase Inhibitors; Lead; Lymphoma; Malignant Neoplasms; Malignant neoplasm of prostate; Murine leukemia virus; Mus; Mutation; Patients; Population; Prevalence; Prostate; Prostate carcinoma; Prostatic Neoplasms; Proteins; Resistance; Testing; Tissues; Variant; Virus; Virus Diseases; Virus Replication; Western Blotting; protein expression; viral detection

We have recently shown that XMRV replication is inhibited by human A3G, human A3F, and murine APOBEC3, suggesting that XMRV does not efficiently replicate and spread in cells that express these host proteins, and may depend on infection of cells that express low levels of these factors. To determine whether XMRV replicates in human PBMCs, we will quantify XMRV replication in PBMCs. We will also quantify the extent of A3 protein expression in normal prostate tissues and prostate carcinomas. To determine the incidence of XMRV in prostate cancer, we will obtain prostate tumors and carry out sensitive PCR assays to determine the extent of XMRV infection. We will also develop ELISA and western blot assays for XMRV detection. To test the hypothesis that co-infection with HIV-1 will lead to the degradation of A3 proteins and allow XMRV to replicate more efficiently, we will determine the incidence of XMRV co-infection in HIV-1 infected individuals with lymphomas and other malignancies. We previously showed that XMRV replication is sensitive to the HIV-1 integrase inhibitor raltegravir. We will carry out mutational analysis of XMRV integrase to identify mutations that confer raltegravir resistance. We will also select for raltegravir-resistant XMRV variants in cell lines and compare the mutations to those selected in HIV-1.

我们最近表明，XMRV 复制受到人 A3G、人 A3F 和鼠 APOBEC3 的抑制，这表明 XMRV 不能在表达这些宿主蛋白的细胞中有效复制和传播，并且可能依赖于表达低水平这些因子的细胞的感染。为了确定 XMRV 是否在人类 PBMC 中复制，我们将量化 PBMC 中的 XMRV 复制。我们还将量化正常前列腺组织和前列腺癌中 A3 蛋白表达的程度。为了确定 XMRV 在前列腺癌中的发生率，我们将获取前列腺肿瘤并进行敏感的 PCR 检测以确定 XMRV 感染的程度。我们还将开发用于 XMRV 检测的 ELISA 和蛋白质印迹测定法。为了检验 HIV-1 合并感染将导致 A3 蛋白降解并使 XMRV 更有效复制的假设，我们将确定患有淋巴瘤和其他恶性肿瘤的 HIV-1 感染个体中 XMRV 合并感染的发生率。我们之前表明 XMRV 复制对 HIV-1 整合酶抑制剂拉替拉韦敏感。我们将对 XMRV 整合酶进行突变分析，以确定导致拉替拉韦耐药的突变。我们还将在细胞系中选择拉替拉韦耐药的 XMRV 变异体，并将这些突变与 HIV-1 中选择的突变进行比较。