Measurement of in vivo mutations in human lymphocytes

A means of measuring in vivo mutations in man would be valuable in assessing the importance of mutation in ageing and human disease and for monitoring individuals exposed to environmental mutagens and carcinogens. The hypoxanthine–guanine phosphoribosyl transferase (HGPRT) locus has been used to stu...

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Bibliographic Details
Published in:Nature (London) 1983-03, Vol.302 (5904), p.155-156
Main Authors: Morley, Alexander A, Trainor, Kevin J, Seshadri, Ram, Ryall, Richard G
Format: Article
Language:English
Subjects:
Cells, Cultured
Clone Cells
Drug Resistance
Humanities and Social Sciences
Humans
Hypoxanthine Phosphoribosyltransferase - genetics
letter
Lymphocytes - ultrastructure
multidisciplinary
Mutation
Science
Science (multidisciplinary)
Selection, Genetic
Thioguanine - pharmacology
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Summary:A means of measuring in vivo mutations in man would be valuable in assessing the importance of mutation in ageing and human disease and for monitoring individuals exposed to environmental mutagens and carcinogens. The hypoxanthine–guanine phosphoribosyl transferase (HGPRT) locus has been used to study mutagenesis in cultured mammalian (and other) cells 1 ; clones are selected by their ability to grow in the presence of the purine analogue, 6-thioguanine (6-TG). Such clones are almost always HGPRT − and are therefore probably the progeny of a mutant cell. This system has been applied to human lymphocytes by using autoradiography to detect single mutant cells 2–6 . This approach probably does detect mutant cells, particularly if their number is increased, but the results obtained are variable and rather imprecise 4,5 . Also, the technique does not allow isolation of clones, thus the mutational origin of thioguanine-resistant (TG r ) cells cannot be determined as the gene product, HGPRT, cannot be measured. Previous methods for cloning peripheral blood lymphocytes (PBL) have been so inefficient that detection of TG r lymphocytes has not been practicable. We have cloned PBL by a technique which is highly efficient (20–60% of PBL form a clone) and results in large clones of 10 3 –10 4 cells that are readily scored using an inverted microscope 6,7 . We report here that the in vivo mutation frequency is of the order of that observed for human fibroblast lines cultured in vitro 1 .
ISSN:0028-0836
1476-4687
DOI:10.1038/302155a0