DIRECT MUTAGENICITY TESTING IN MAN

人类直接突变性测试

• 批准号: 3169367
• 负责人: RICHARD J ALBERTINI
• 金额: $ 17.82万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-09-01 至 1988-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3169367
• 关键词: 6 thioguanine; T lymphocyte; alveolar macrophages; autoradiography; biopsy; blood /lymphatic neoplasm; breast neoplasm /cancer diagnosis; breast neoplasms; cancer risk; carcinogen testing; cell transformation; clone cells; congenital disorders; drug related neoplasm /cancer; drug resistance; early diagnosis; environment related neoplasm /cancer; enzyme linked immunosorbent assay; gene mutation; genetic markers; human population genetics; human subject; hypoxanthine phosphoribosyltransferase; immunogenetics; immunosuppression; longitudinal human study; lymphocyte; macrophage; multiple primary neoplasia; mutagen testing; mutagens; mutant; neoplasm /cancer chemotherapy; neoplasm /cancer diagnosis; neoplasm /cancer genetics; neoplasm /cancer immunotherapy; neoplasm /cancer radiation therapy; phototherapy; radiation genetics; radiotracer; skin neoplasms; sperm; ultraviolet radiation

This research project will investigate and improve the recently described 6-thioguanine resistant (TGr) T-lymphocyte (T-Ly) clonal assay for detecting somatic gene mutations occurring in vivo in humans in order to: (i) provide assays for human in vivo mutagenicity monitoring and (ii) define the structural and molecular bases of these mutations. The clonal assay will be made more precisely quantitative and then be used to define the range of TGr T-Ly mutant frequency values for normal individuals and for persons exposed to mutagens. Wild type and TGr T-Ly colonies, recovered from clonal assays, will be characterized as to their phenotypic stability, gene product alterations, T-Ly surface antigen subclass, chromosome change, DNA structural alteration in the hprt gene, alterations in specific messenger RNA and specific configeration of the gene for the T-cell receptor. These characteristics will allow definition of the spectrum of hprt gene mutations occurring in vivo in humans, will provide minimal estimates as to the number of mutations responsible for an observed collection of mutants, and will determine if selected mutant characteristics differentiate between "spontaneous" and "induced" in vivo somatic gene mutations in humans. The older autoradiographic assay for quantitating TGr T-Ly variant frequencies in human peripheral blood will be refined, used for human studies, and results obtained using it will be compared with results obtained for similar blood samples using the clonal assay. The T-Ly clonal assay will then be used as the basis for developing a multi-locus (i.e. HLA-loss) somatic mutation detection system that will be capable of detecting alteration of autosomal genes in vivo in humans. Wild type and HLA mutant T-Ly colonies recovered from these assays will be characterized in a manner analogous to the characterization of hprt mutants.

这项研究项目将调查和改进最近描述的 抗6-硫鸟嘌呤(TGR)T淋巴细胞(T-Ly)克隆性试验 检测人体体内发生的体细胞基因突变，以便： (I)提供人体体内诱变性监测的化验方法；及(Ii) 确定这些突变的结构和分子基础。克隆人 化验将更精确地定量，然后用于定义 正常人TGR T-Ly突变频率值范围 对于暴露在诱变剂中的人。野生型和TGR T-Ly菌落， 从克隆分析中恢复的，将被表征为它们的表型 稳定性，基因产物改变，T-Ly表面抗原亚类， 染色体改变，HPRT基因的DNA结构改变，改变 在特定的信使RNA和基因的特定配置中 T细胞受体。这些特征将允许定义 在人体内发生的HPRT基因突变谱，将提供 对导致观察到的基因突变数量的最小估计 突变的集合，并将确定所选的突变 体内“自发”和“诱导”的特征区别 人类的体细胞基因突变。较旧的放射自显影分析 定量检测人外周血中TGR T-Ly变异频率将是 精炼，用于人体研究，使用它获得的结果将被 与使用克隆血样获得的类似血液样本的结果进行比较 化验。然后，T-Ly克隆试验将作为开发的基础 一种多位点(即HLA缺失)体细胞突变检测系统，将 能够检测到人体内常染色体基因的变化。 从这些检测中回收的野生型和人类白细胞抗原突变T-Ly克隆将是 其特征类似于HPRT突变体的特征。