DIRECT MUTAGENICITY TESTING IN MAN

该研究计划的目标是开发和验证 人体体内体细胞突变的研究方法。是这样的 化验对于遗传风险评估以及 回答有关分子性质的基本问题 人类的突变。对突变机制的了解有 这对理解人类疾病具有重要意义。这些 研究的目的是充分描述“自发”基因突变的特征 在活体内发生在人类淋巴细胞中，并将其与自发性进行比较 用模型烷化剂在体内诱发基因突变 在人类淋巴细胞中。这些目标将通过以下方式实现 实现以下具体目标：(1)完成人口 次黄嘌呤-鸟嘌呤磷酸核糖转移酶的体内研究 正常人外周血T淋巴细胞HPRT基因突变研究 人类，以及定义T细胞克隆的体内扩增 在正常人中，被确定为关于以下方面的“异常值” HPRT的突变频率值(&gt；50×10-6)和克隆性 变种人。(2)野生型和HPRT突变型T细胞的鉴定 从正常人外周血中恢复的克隆 表面标志表型，特异性T细胞受体(TRC)基因 重排模式和HPRT基因改变。(3)量化 L-苯丙氨酸芥末诱变hprt的特征 多发性骨髓瘤患者外周血中淋巴细胞的体内研究 骨髓瘤接受这种治疗的骨髓瘤(4)量化和 人外周血中淋巴细胞HPRT突变的体内特征 乳腺癌或骨髓瘤患者的淋巴引流和 将结果与从外周血中获得的结果进行比较 以比较分裂细胞和非分裂细胞中的突变。 (5)建立一种用于定量和表征的克隆检测方法 人类常染色体人类白细胞抗原基因在活体内的突变 淋巴细胞，用于比较和与X-连锁的HPRT基因座 突变频率的结果。(6)建立克隆性检测方法 体内发生的基因突变的量化和表征 人类B淋巴细胞，包括对克隆性的定义 通过免疫球蛋白(Ig)基因重排模式进行比较 与T淋巴细胞HPRT基因座检测结果一致。

The objective of this research program is to develop and validate methods for studying in vivo somatic cell mutation in humans. Such assays are necessary for genetic risk estimates, as well as answering basic questions concerning the molecular nature of mutation in humans. Knowledge of the mechanism of mutation has important implications for understanding human disease. These studies aim to fully characterize "spontaneous" gene mutation occurring in vivo in human lymphocytes, and compare "spontaneous" with model alkylating agent induced gene mutation occurring in vivo in human lymphocytes. These objectives will be attained by achieving the following specific aim: (1) Completing population studies of in vivo hypoxanthine-guanine phosphoribosyltransferase gene (hprt) mutation in peripheral blood T-lymphocytes in normal humans, as well as defining in vivo amplifications of T-cell clones in normal individuals, ascertained as "outliers" with respect to mutant frequency values (> 50 x 10-6) and clonality of hprt mutants. (2) Characterizing wild type and hprt mutant T-cell clones recovered from the peripheral blood of normal individuals of surface marker phenotypes, specific T-cell receptor (TRC) gene rearrangement patterns, and hprt gene alterations. (3) Quantifying and characterizing L-phenylalanine mustard induced hprt mutation in vivo in peripheral blood lymphocytes of patients with multiple myeloma receiving such treatment. (4) Quantifying and characterizing in vivo hprt mutation in lymphocytes obtained from draining lymph nodes from breast cancer or myeloma patients and comparing findings with those obtained from peripheral blood in order to compare mutation in dividing versus non-dividing cells. (5) Developing a clonal assay for quantifying and characterizing gene mutations arising in vivo at the autosomal HLA locus in human lymphocytes, for comparison and with the X-linked hprt locus mutation frequency results. (6) Developing a clonal assay for quantifying and characterizing gene mutations arising in vivo in human B-lymphocytes, including definition of clonality as reflected by immunoglobulin (Ig) gene rearrangement patterns, for comparison with the T-lymphocyte hprt locus results.