title: The human T-cell cloning assay: identifying genotypes susceptible to drug toxicity and somatic mutation.
abstract: Humans exhibit marked genetic polymorphisms in drug metabolism that contribute to high incidence of adverse effects in susceptible individuals due to altered balance between metabolic activation and detoxification. The T-cell cloning assay, which detects mutations in the gene for hypoxanthine-guanine phosphoribosyl transferase (HPRT), is the most well-developed reporter system for studying specific locus mutation in human somatic cells. The assay is based on a mitogen- and growth factor-dependent clonal expansion of peripheral T-lymphocytes in which the 6-thioguanine-resistant HPRT mutants can be selected, enumerated, and collected for molecular analysis of the mutational nature. The assay provides a unique tool for studying in vivo and in vitro mutagenesis, for investigating the functional impact of common polymorphism in metabolism and repair genes, and for identifying risk genotypes for drug-induced toxicity and mutagenicity. This chapter presents a simple and reliable method for the enumeration of HPRT mutant frequency induced in vitro without using any source of recombinant interleukin-2. The other main feature is that only truly induced and unique mutants are collected for further analysis.
authors.affiliation: Department of Biosciences, Karolinska Institute, CNT/NOVUM, 141 57, Huddinge, Sweden, saimei.hou@cnt.ki.se.
authors.forename: Sai-Mei
authors.initials: SM
authors.lastname: Hou
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D004252:DNA Mutational Analysis; D017930:Genes, Reporter; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D009153:Mutagens / Q000633:toxicity; D009154:Mutation; D012306:Risk; D013601:T-Lymphocytes / Q000502:physiology*
chemical_list: D009153:Mutagens; D007041:Hypoxanthine Phosphoribosyltransferase