Page T M, Broock R L, Nyhan W L, Nieto L H
Clin Chim Acta. 1986 Feb 15;154(3):195-201. doi: 10.1016/0009-8981(86)90031-8.
Growth properties of cultured fibroblasts in selective media were used to characterize the HPRT enzyme of a patient with a new variant of hypoxanthine phosphoribosyl transferase (HPRT) with deficient activity. The clinical phenotype of the patient was typical of the Lesch-Nyhan syndrome. However, cells of the patient were not selected for by growth in either 8-azaguanine or 6-thioguanine. Assay of the activity of the enzyme in erythrocyte lysates revealed values of approximately zero, while in the intact fibroblast assay the level of activity was 1.4% of normal. The heterozygous mother of the patient, unlike heterozygotes for the classic Lesch-Nyhan enzyme, had a level of activity in erythrocyte lysates that was 45% of control. In the presence of selective agents in vitro the cells of the patient retained sufficient HPRT activity to permit a degree of toxicity indistinguishable from that observed in normal cells although the degree of the deficiency was so great that it led to the complete Lesch-Nyhan phenotype. These findings call into question the use of selective agents for the identification of HPRT- cells in the detection of heterozygosity.
利用培养的成纤维细胞在选择性培养基中的生长特性来表征一名患有次黄嘌呤磷酸核糖基转移酶(HPRT)新变体且活性不足的患者的HPRT酶。该患者的临床表型为典型的莱施-奈恩综合征。然而,患者的细胞在8-氮杂鸟嘌呤或6-硫鸟嘌呤中均未通过生长筛选。红细胞裂解物中该酶活性的测定显示值约为零,而在完整成纤维细胞测定中,活性水平为正常水平的1.4%。患者的杂合子母亲与经典莱施-奈恩酶的杂合子不同,其红细胞裂解物中的活性水平为对照的45%。在体外存在选择性试剂的情况下,患者的细胞保留了足够的HPRT活性,以产生与正常细胞中观察到的毒性程度无法区分的毒性程度,尽管缺陷程度很大,导致了完全的莱施-奈恩表型。这些发现对在检测杂合性时使用选择性试剂来鉴定HPRT缺陷细胞提出了质疑。