Abstract
In females heterozygous for the Lesch-Nyhan (LN) mutation, there is mosaicism of peripheral blood lymphocytes (PBLs) with regard to sensitivity to 6-thioguanine (TG) inhibition of tritiated thymidine ([3H]Tdr) incorporation following phytohemagglutinin (PHA) stimulation. That there are two populations of PBLs, normal and mutant (LN-like), has been demonstrated by an autoradiographic enumerative assay. A single three-generation family containing six potentially heterozygous females was studied. Five of the six were mosaics with frequencies of TG-resistant (TGr) PBLs ranging from 1.4×10−3 to 4.2×10−3 when tested at 2×10−4 m TG. The median frequency of TGr PBLs in 63 healthy non-LN individuals between the ages of 11 and 75 years was found to be 1.1×10−4 (mean 1.3×10−4) (10th and 90th percentiles—6.1×10−5 and 2.1×10−4) and was not age related. The sixth potentially heterozygous female in the current family had a TGr PBL frequency of 1.9×10−4. In the five females with elevated TGr PBL frequencies, TGr skin fibroblasts with frequencies ranging from 26% to 100% of the sample tested were found; in the female with the normal TGr PBL frequency, no TGr skin fibroblasts were found. The former group was considered to be LN heterozygous. Four of the five had been previously diagnosed as such. The latter individual is considered to be genotypically normal. Females who are heterozygotes for the LN mutation have two populations of PBLs.
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References
Albertini, R. J., and DeMars, R. (1970). Diploid azaguanine-resistant mutants of cultured human fibroblasts. Science 169482.
Albertini, R. J., and DeMars, R. (1973). Detection and quantification of X-ray induced mutation in cultured, diploid human fibroblasts. Mutation Res. 18199.
Albertini, R. J., and DeMars, R. (1974). Mosaicism of peripheral blood lymphocyte populations in females heterozygous for the Lesch-Nyhan mutation. Biochem. Genet. 11397.
Allison, A. C., Hovi, T., Watts, R. W. E., and Webster, A. B. D. (1977). The role of de novo purine synthesis in lymphocyte transformation. In Elliot, K., and Fitzsimons, D. A. (eds.), Purine and Pyrimidine Metabolism, Elsevier/North-Holland Excerpta Medica, Amsterdam, p. 207.
Arley-Nelson, J., Carpenter, J. W., Rose, L. M., and Adamson, D. J. (1975). Mechanisms of action of 6-thioguanine, 6-mercaptopurine and 8-azaguanine. Cancer Res. 353872.
Bakay, B., Nyhan, W. L., Fawcett, N., and Kogut, M. D. (1972). Isoenzymes of hypoxanthineguanine phosphoribosyltransferase in a family with partial deficiency of the enzyme. Biochem. Genet. 773.
Beutler, E., Yeh, M., and Fairbanks, V. F. (1962). The normal human female as a mosaic of X-chromosome activity: Studies using the gene for glucose-6-phosphate dehydrogenase deficiency as a marker. Proc. Natl. Acad. Sci. 489.
Cox, R. P., Krauss, M. R., Balis, M. E., Yip, L. C., Jansen, V., and Dancis, J. (1974). Incorporation of hypoxanthine by PHA-stimulated HG-PRT-deficient lymphocytes. Exp. Cell Res. 88289.
Dancis, J., Berman, P. H., Jensen, V., and Balis, M. E. (1968). Absence of mosaicism in the lymphocyte in X-linked congenital hyperuricosuria. Life Sci. 7587.
Davidson, R. G., Nitowisky, H. M., and Childs, B. (1963). Demonstration of two populations of cells in the human female heterozygous for glucose-6-phosphate dehydrogenase variants. Proc. Natl. Acad. Sci. 50481.
deBruyn, C. H. M. M., Oei, T. L., and terHaar, B. G. A. (1974). Studies on hairroots for carrier detection on hypoxanthine-guanine phosphoribosyltransferase deficiency. Clin. Genet. 5449.
DeMars, R. (1971). Genetic studies of HG-PRT deficiency and the Lesch-Nyhan syndrome with cultured human cells. Fed. Proc. 30944.
DeMars, R., Sarto, G., Felix, J. S., and Benke, P. (1969). LN mutation: Prenatal detection with amniotic fluid cells. Science 1641303.
Elion, G. B. (1967). Biochemistry and pharmacology of purine analogues. Fed. Proc. 26898.
Elion, G. B., and Hitchings, G. H. (1965). Metabolic basis for the actions of analogs of purines and pyrimidines, In Golden, A., Hawkins, F., and Schnitzer, R. J. (eds.), Advances in Chemotherapy 2, Academic Press, New York, p. 91.
Emmerson, B. T., and Wyngaarden, J. B. (1969). Purine metabolism in heterozygous carriers of HG-PRT deficiency. Science 1661533.
Felix, J. S., and DeMars, R. (1971). Detection of females heterozygous for the LN mutation by 8-azaquanine-resistant growth of cultured fibroblasts. J. Lab. Clin. Med. 27596.
Francke, U., Bakay, B., and Nyhan, W. L. (1973). Detection of heterozygous carriers of the Lesch-Nyhan syndrome by electrophoresis of hairroot lysates. J. Pediatr. 82472.
Fujimoto, W. Y., and Seegmiller, J. E. (1970). HG-PRT deficiency: Activity in normal, mutant and heterozygote-cultured human skin fibroblasts. Proc. Natl. Acad. Sci. 65577.
Gallop, P. M., and Paz, M. A. (1978). Protein Modification in Aging Cells. Sci. Birth Defects Orig. Art. Ser. 14(1403.
Gartler, S. M., and Andina, R. J. (1976). Mammalian X-chromosome inactivation. In Harris, H., and Hirschhorn, K. (eds.), Human Genetics, Vol. 7, Plenum Press, New York, p. 99.
Hovi, T., Allison, A. C., Raivio, K. O., and Vaheri, A. (1977). Purine metabolism and control of cell proliferation. In Elliot, K., and Fitzsimons, D. W. (eds.), Purine and Pyramidine Metabolism, Elsevier/North-Holland Excerpta Medica, Amsterdam, p. 207.
Kelley, W. N. (1968). Ezymology and biochemistry A: HG-PRT deficiency in the Lesch-Nyhan syndrome and gout. Fed. Proc. 271047.
Lesch, M., and Nyhan, W. L. (1964). A famial disorder of uric acid metabolism and central nervous system function. Am. J. Med. 36561.
Ling, N. R., and Kay, J. E. (1975). DNA synthesis and the cell cycle. In Lymphocyte Stimulation, Chap. 14, North-Holland, Amsterdam, p. 357.
Lyons, M. G. (1961). Gene action in the X-chromosome of the mouse (Mus musculus L.). Nature 190372.
McDonald, J. A., and Kelley, W. N. (1972). LN syndrome: Absence of the mutant enzyme in erythrocytes of a heterozygote for both normal and mutant HGPRT. Biochem. Genet. 621.
Miech, R. P., Parks, R. E. Jr., Anderson, J. H., Jr., and Sartorelli, A. C. (1967). An hypothesis on the mechanism of action of 6-thioguanine. Biochem. Pharmacol. 162222.
Migeon, B., DerKaloustian, V. M., Nyhan, W. L., Young, W. J., and Childs, B. (1968). X-linked HG-PRT deficiency: Heterozygote has two clonal populations. Science 160425.
Migeon, B. R. (1970). X-linked HG-PRT deficiency: Detection of heterozygotes by selective medium. Biochem. Genet. 4377.
Nyhan, W. L., Bakay, B., Connor, J. D., Marks, J. F., and Keele, D. K. (1970). Hemizygous expression of heterozygotes for the Lesch-Nyhan syndrome. Proc. Natl. Acad. Sci. 65214.
Rosenbloom, F. M., Kelly, W. N., Henderson, J. F., and Seegmiller, J. E. (1967). Lyon hypothesis and X-linked disease. Lancet 2305.
Salzmann, J., DeMars, R., and Benke, P. (1968). Single allele expression at an X-linked hyperuricemia locus in heterozygous human cells. Proc. Natl. Acad. Sci. 60545.
Sanford, K., Earle, W. R., Evans, J., Waltz, H. K., and Shannon, J. E. (1950). The measurement of proliferation in tissue cultures by enumeration of cell nuclei. J. Natl. Cancer Inst. 11773.
Schellekens, P. T. A., and Eijsvoogel, V. P. (1968). Lymphocyte transformation in vitro: Tissue culture conditions in quantitative measurements. Clin. Exp. Immunol. 3571.
Scholar, E. M., and Calabresi, P. (1973). Identification of the enzymatic pathways of nucleotide metabolism in human lymphocytes and leukemia cells. Cancer Res. 8394.
Schwarzmeier, J. D., and Maser, K. (1972). Studies on the synthesis de novo of purines in normal, PHA stimulated and leukemic lymphocytes. In Gerlach, E. (ed.), Erythrocytes, Thrombocytes and Leukocytes, 2nd International Symposium, Thiere-Verlag, Stuttgart, p. 419.
Seegmiller, J. E., Rosenbloom, F. M., and Kelley, W. M. (1967). Enzyme defect associated with a sex-linked human neurological disorder and excessive purine synthesis. Science 1551682.
Soren, L. (1973). Variability of the time at which PHA-stimulated lymphocytes initiate DNA synthesis. Exp. Cell Res. 78201.
Sperling, L., Eliam, G., Schmidt, R., Mundel, G., and deVries, A. (1971). Purine base incorporation into erythrocytes nucleotides in HG-PRT deficiency. Biochem. Med. 5173.
Strauss, G. H., and Albertini, R. M. (1977). 6-Thioguanine resistant lymphocytes in human peripheral blood. In Scott, D., Bridges, B. A., and Sobels, F. H. (eds.), Progress in Genetic Toxicology, Elsevier/North-Holland, Amsterdam, p. 327.
Strauss, G. H., and Albertini, R. J. (1979). Enumeration of 6-thioguanine resistant peripheral blood lymphocytes in man as a potential test for somatic cell mutations arising in vivo. Mutation Res. 61353.
Strauss, G. H., Albertini, R. J., Krusinski, P. A., and Baughman, R. D. (1979). 6-Thioguanine resistant peripheral blood lymphocytes in humans following psoralen, long-wave ultraviolet light (PUVA) therapy. J. Invest. Dermatol. (in press).
Wood, S., and Pinsky, L. (1970). Lesch-Nyhan syndrome, rapid detection of heterozygotes. Clin. Genet. 1216.
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This work was supported by National Institutes of Health Grant No. PHS RO1 15450 and the Surgical Associates Foundation, Inc., University of Vermont College of Medicine. Elizabeth F. Allen was supported by the NIH National Research Service Award No. 5 F32 CA 05918.
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Strauss, G.H., Allen, E.F. & Albertini, R.J. An enumerative assay of purine analogue resistant lymphocytes in women heterozygous for the Lesch-Nyhan mutation. Biochem Genet 18, 529–547 (1980). https://doi.org/10.1007/BF00484400
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DOI: https://doi.org/10.1007/BF00484400