Abstract
Phytohemagglutinin-stimulated thymidine-H3 incorporation into peripheral blood lymphocytes in vitro was inhibited by 8-azaguanine (AG) in cells from normal individuals, but not in cells from boys with the Lesch-Nyhan (L-N) syndrome. Artificial cell mixtures in which the AG-resistant minority L-N population comprised more than 1.0% of the total were clearly differentiated from normals by this technique. The method was used to search for blood cell mosaicism in females heterozygous for the X-chromosomal L-N mutation. In three adolescent girls, a minority population of AG-resistant lymphocytes comprising 5–10% of the total was found, while no such mosaicism was detected in a heterozygous woman of 40. Although blood cell mosaicism had not previously been found in female carriers of the L-N mutation, the sensitivity of the present method exceeds that of previous studies. Our findings suggest that in vivo selection against lymphocytes with the L-N mutation does occur but is not absolute and may be age related. The present method has applications in studying this phenomenon, and, with modification, may provide a direct assay system to detect the in vivo frequencies of AG-resistant lymphocytes.
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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.
Bakay, B., Nyhan, W. L., Fawcett, N., and Kogut, M. D. (1972). Isoenzymes of hypoxanthine-guanine phosphoribosyltransferase in a family with partial deficiency of the enzyme. Biochem. Genet. 773.
Balis, M. E. (1968). Enzymology and biochemistry B. Aspects of purine metabolism. Fed. Proc. 271067.
Dancis, J., Berman, P. H., Jansen, V., and Balis, M. E. (1968). Absence of mosaicism in the lymphocyte in X-linked congenital hyperuricemia. Life Sci. 7587.
Dancis, J., Cox, R. P., Berman, P. H., Jansen, V., and Balis, M. E. (1969). Cell population density and phenotypic expression of tissue culture fibroblasts from heterozygotes of Lesch-Nyhan's disease (inosinate-pyrophosphorylase deficiency). Biochem. Genet. 3609.
Davidson, J. D., and Winter, T. S. (1964). Purine nucleotide pyrophosphorylases in 6-mercaptopurine-sensitive and resistant human leukemias. Cancer Res. 24261.
DeMars, R. (1971). Genetic studies of HG-PRT deficiency and the Lesch-Nyhan syndrome with cultured human cells. Fed. Proc. 30944.
DeMars, R., and Held, K. (1972). The spontaneous azaguanine-resistant mutants of diploid human fibroblasts. Humangenetik 1687.
DeMars, R., and Nance, W. E. (1964). Electrophoretic variants of glucose-6-phosphate dehydrogenase and the single-active-X in cultivated human cells. In Defendi, V. (ed.), Retention of Functional Differentiation in Cultured Cells, Wistar Institute Press, Philadelphia, pp. 35–48.
Emmerson, B. T., and Wyngaarden, J. B. (1969). Purine metabolism in heterozygous carriers of hypoxanthine-guanine phosphoribosyltransferase deficiency. Science 1661533.
Felix, J. S. (1971). Genetic studies on cultured cells bearing the Lesch-Nyhan mutation (hypoxanthine-guanine phosphoribosyltransferase deficiency) with attempts to derepress the inactive X-chromosome in heterozygous female cells. Ph.D. thesis, University of Wisconsin.
Felix, J. S., and DeMars, R. (1971). Detection of females heterozygous for the Lesch-Nyhan mutation by 8-azaguanine-resistant growth of cultured fibroblasts. J. Lab. Clin. Med. 27596.
Fujimoto, W. Y., and Seegmiller, J. E. (1970). Hypoxanthine-guanine phosphoribosyltransferase deficiency: Activity in normal, mutant and heterozygote-cultured human skin fibroblasts. Proc. Natl. Acad. Sci. 65577.
Fujimoto, W. Y., Subak-Sharpe, J. H., and Seegmiller, J. E. (1971). HG-PRT deficiency: Clinical agents selective for mutant or normal cultured fibroblasts in mixed and heterozygous cultures. Proc. Natl. Acad. Sci. 681516.
Goldstein, J. L., Marks, J. F., and Gartler, S. M. (1971). Expression of two X-linked genes in human hair follicles of double heterozygotes. Proc. Natl. Acad. Sci. 681425.
Ham, R. G. (1963). An improved nutrient solution for Chinese hamster and human cell lines. Exptl. Cell Res. 29515.
Henderson, J. F., Kelley, W. N., Rosenbloom, F. M., and Seegmiller, J. E. (1969). Inheritance of purine phosphoribosyltransferase in man. Am. J. Hum. Genet. 2161.
Kelley, W. N. (1968). Enzymology and biochemistry A. HG-PRT deficiency in the Lesch-Nyhan syndrome and gout. Fed. Proc. 271047.
Kelley, W. N., and Wyngaarden, J. B. (1972). In Stanbury, J. B., Wyngaarden, J. B., and Frederickson, D. S. (eds.), The Metabolic Basis of Inherited Disease, McGraw-Hill, New York, pp. 969–991.
Kelley, W. N., Rosenbloom, F. M., Miller, J., and Seegmiller, J. E. (1968). An enzymatic basis for variation in response to allopurinol. New Engl. J. Med. 278287.
Kelley, W. N., Greene, M. L., Rosenbloom, F. M., Henderson, J. F., and Seegmiller, J. E. (1969). Review: HG-PRT-deficiency in gout. Ann. Int. Med. 70155.
Lesch, M., and Nyhan, W. (1964). A familial disorder of uric acid metabolism and central nervous system function. Am. J. Med. 36561.
Luria, S. E., and Delbruck, M. (1943). Mutations of bacteria from virus sensitivity to virus resistance. Genetics 28491.
Lyon, M. F. (1961). Gene action in the X-chromosome of the mouse (Mus musculus L.). Nature 190372.
McDonald, J. A., and Kelley, W. N. (1972). Lesch-Nyhan syndrome: Absence of the mutant enzyme in erythrocytes of a heterozygote for both normal and mutant hypoxanthine-guanine phosphoribosyltransferase. Biochem. Genet. 621.
Migeon, B. R., Der Kaloustian, V. M., Nyhan, W. L., Young, W. J., and Childs, B. (1968). Hypoxanthine-guanine phosphoribosyltransferase deficiency: Heterozygote has two clonal populations. Science 16425.
Nyhan, W. L. (1968a). Clinical features of the Lesch-Nyhan syndrome. Introduction—Clinical and genetic features. Seminars on the Lesch-Nyhan syndrome. Fed. Proc. 271027.
Nyhan, W. L. (1968b). Clinical features of the Lesch-Nyhan syndrome. Summary of clinical features. Seminars on the Lesch-Nyhan syndrome. Fed. Proc. 271033.
Nyhan, W. L., Pesek, J., Sweetman, L., Carpenter, D. G., and Carter, C. H. (1967). Genetics of an X-linked disorder of uric acid metabolism and cerebral function. Pediat. Res. 15.
Nyhan, W. L., Bakay, B., Connor, J. D., Marks, J. F., and Keele, D. K. (1970). Hemizygous expression of glucose-6-phosphate dehydrogenase in erythrocytes of heterozygotes for the Lesch-Nyhan syndrome. Proc. Natl. Acad. Sci. 65214.
Rabinowitz, Y. (1964). Separation of lymphocytes, polymorphonuclear leukocytes and monocytes on glass columns, including tissue culture observations. Blood 23811.
Rosenbloom, F. M., Kelley, W. N., Henderson, J. F., and Seegmiller, J. E. (1967). Lyon hypothesis and X-linked disease. Lancet 2305.
Russell, L. B. (1963). Mammalian X-chromosome action: Inactivation limited in spread and in region of origin. Science 40976.
Salzmann, J., DeMars, R., and Benke, L. (1968). Single allele expression at an X-linked hyperuricemia locus in heterozygous human cells. Proc. Natl. Acad. Sci. 60545.
Seegmiller, J. E., Rosenbloom, F. M., and Kelley, W. N. (1967). Enzyme defect associated with a sex-linked human neurological disorder and excessive purine synthesis. Science 1551682.
Shapiro, S. L., Sheppard, G. L., Dreifuss, F. E., and Newcombe, D. S. (1966). X-linked recessive inheritance of a syndrome of mental retardation with hyperuricemia. Proc. Soc. Exptl. Biol. Med. 122609.
Sørensen, S. F., Anderson, V., and Giese, J. (1969). Studies on the quantitation of the lymphocyte response in vitro. Acta Pathol. Microbiol. Scand. 76259.
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Albertini, R.J., DeMars, R. Mosaicism of peripheral blood lymphocyte populations in females heterozygous for the Lesch-Nyhan mutation. Biochem Genet 11, 397–411 (1974). https://doi.org/10.1007/BF00486413
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DOI: https://doi.org/10.1007/BF00486413