Abstract
Lesch-Nyhan syndrome is caused by a severe genetic deficiency of hypoxanthine phosphoribosyltransferase (HPRT) and is characterized by central nervous system disorders, gout, and in some cases, macrocytic anemia. Women heterozygous for HPRT deficiency are healthy but their somatic cells are mosaic for enzyme deficiency owing to random inactivation of the X chromosome. Frequencies of red blood cells and T cells deficient in HPRT are significantly lower than the expected 50% in heterozygotes, suggesting that HPRT-negative blood cells are selected against in heterozygotes. To determine at which stage of hematopoiesis such selection occurs, we determined the frequencies of HPRT-negative T, B and erythroid precursor cells in three heterozygotes. Since the cloning efficiencies of T and B cells and colony forming efficiency of burst-forming unit erythroid (BFU-E) for sample from Lesch-Nyhan patients were similar to those of normal cells, HPRT deficiency does not seem to render the differentiated cells less efficient for proliferation. However, the frequencies of HPRT-negative T and B cells, and BFU-E were all less than 10% in each of the three heterozygotes. Although the frequencies of HPRT-negative cells showed tenfold variations between the heterozygotes, each heterozygote had similar frequencies of HPRT-negative cells in the three cell types. These results suggest that HPRT is important at early stages of hematopoiesis, but less so after the cells have differentiated into T cells, B cells and erythroid precursor cells.
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Albertini RJ, Castle KL, Borcherding WR (1982) T-cell cloning to detect the mutant 6-thioguanine-resistant lymphocytes present in human peripheral blood. Proc Natl. Acad Sci USA 79:6617–6621
Albertini RJ, O'Neill JP, Nicklas JA, Heintz NH, Kelleher P (1985) Alterations of the hprt gene in human in vivo-derived 6-thioguanine-resistant T lymphocytes. Nature 316:369–371
Allison AC, Watts WE, Hovi T, Webster ADB (1975) Immunological observations on patients with Lesch-Nyhan syndrome, and on the role of de-novo purine synthesis in lymphocyte transformation. Lancet II:1179–1182
Bradley WEC, Gareau JLP, Seifert AM, Messing K (1987) Molecular characterization of 15 rearrangements among 90 human in vivo somatic mutants shows that deletions predominate. Mol Cell Biol 7:956–960
Davies MJ, Lovell DP, Anderson D (1992) Thioguanine-resistant mutant frequency in T-lymphocytes from a healthy human population. Mutat Res 265:165–171
Dempsey JL, Morley AA, Seshadri RS, Emmerson BT, Gordon R, Bhagat CI (1983) Detection of the carrier state for an X-linked disorder, the Lesch-Nyhan syndrome, by the use of lymphocyte cloning. Hum Genet 64:288–290
Dempsey JL, Seshadri RS, Morley AA (1985) Increased mutation frequency following treatment with cancer chemotherapy. Cancer Res 45:2873–2877
Francke U, Bakay B, Nyhan WL (1973) Detection of heterozygous carriers of the Lesch-Nyhan syndrome by electrophoresis of hair root lysates. J Pediatr 82:472–478
Gartler SM, Scott RC, Goldstein JL, Campbell B (1971) Lesch-Nyhan syndrome: rapid detection of heterozygotes by use of hair follicles. Science 172:572–574
Gelfrand EW, Fox IH, Stuckey M, Dosch HM (1978) Normal Blymphocyte function in patients with Lesch-Nyhan syndrome and HGPRT deficiency. Clin Exp Immunol 31:205–208
Hakoda M, Akiyama M, Kyoizumi S, Awa AA, Yamakido M, Otake M (1988a) Increased somatic cell mutant frequency in atomic bomb survivors. Mutat Res 201:39–48
Hakoda M, Akiyama M, Kyoizumi S, Kobuke K, Awa AA, Yamakido M (1988b) Measurement of in vivo HGPRT-deficient mutant cell frequency using a modified method for cloning human peripheral blood T-lymphocytes. Mutat Res 197:161–169
Hakoda M, Hirai Y, Kusunoki Y, Akiyama M (1989a) Cloning of in vivo-derived thioguanine-resistant human B cells. Mutat Res 210:29–34
Hakoda M, Hirai Y, Kyoizumi S, Akiyama M (1989b) Molecular analyses of in vivo hprt mutant T cells from atomic bomb survivors. Environ Mol Mutagen 13:25–33
Hakoda M, Hirai Y, Shimba H, Kusunoki Y, Kyoizumi S, Kodama Y, Akiyama M (1989c) Cloning of phenotypically different human lymphocytes originating from a single stem cell. J Exp Med 169:1265–1276
Hakoda M, Nishioka K, Kamatani N (1990) Homozygous deficiency at autosomal locus aprt in human somatic cells in vivo induced by two different mechanisms. Cancer Res 50:1738–1741
Hirota H, Kubota M, Hashimoto H, Adachi S, Matsubara K, Kuwakado K, Akiyama Y, Tsutsui T, Mikawa H (1993) Analysis of hprt gene mutation following anti-cancer treatment in pediatric patients with acute leukemia. Mutat Res 319:113–120
Johnson LA, Gordon RB, Emmerson BT (1976) Two populations of heterozygote erythrocytes in moderate hypoxanthine guanine phosphoriboxyltransferase deficiency. Nature 264:172–174
Kamatani N, Yamanaka H, Nishioka K, Nakamura T, Nakano K, Tanimoto K, Mizuno T, Nishida Y (1984) A new method for the detection of Lesch-Nyhan heterozygotes by peripheral blood T-cell culture using T-cell growth factor. Blood 63:912–916
Lyon MF (1961) Gene action in the X-chromosome of the mouse (Mus musculus L). Nature 190:372–373
McDonald JA, Kelley WN (1972) LN syndrome: absence of the mutant enzyme in erythrocytes of a heterzygote for both normal and mutant hypoxanthine-guanine phosphoribosyltransferase. Biochem Genet 6:21–26
McKeran RO, Howell A, Andrews TM, Watts RWE, Arlett CF (1974) Observation on the growth in vitro of myeloid progenitor cells and fibroblasts from hemizygotes and heterozygotes for “complete” and “partial” hypoxanthine-guanine phosphoribosyltransferase (HGPRT) deficiency, and their relevance to the pathogenesis of brain damage in the Lesch-Nyhan syndrome. J Neurol Sci 22:183–195
Migeon BR, Der Kaloustian VM, Nyhan WL, Young WJ, Childs B (1968) X-linked hypoxanthine-guanine phosphoribosyltransferase deficiency: heterozygote has two clonal populations. Science 160:425–427
Messing K, Bradley WEC (1985) In vivo mutant frequency rises among breast cancer patients after exposure to high doses of gamma radiation. Mutat Res 152:107–112
Metcalf D (1984) Clonal culture of hematopoietic cells: techniques and applications. Elsevier, Amsterdam
Morley AA, Trainor KJ, Seshadri R, Ryall RJ (1983) Measurement of in vivo mutations in human lymphocytes. Nature 302:155–156
Nicklas FA, O'Neill JP, Albertini RJ (1986) Use of T-cell receptor gene probes to quantify the in vivo hprt mutations in human T-lymphocytes. Mutat Res 173:67–72
Nicklas FA, O'Neill JP, Sullivan LML, Hunter TC, Allegretta M, Chastenay BF, Libbus BL, Albertini RJ (1988) Molecular analysis of in vivo hypoxanthine-guanine phosphoribosyltransferase mutations in human T-lymphocytes: II. Demonstration f a clonal amplification of hprt mutant T-lymhpocytes in vivo. Environ Mol Mutagen 12:271–284
Nyhan WL, Bakay B, Connor JD, Marks JF, Keele DK (1970) Hemizygous expression of glucose-6-phoshate dehydrogenase in erythrocytes of hererozygotes for the Lesch-Nyhan syndrome. Proc Natl Acad Sci USA 65:214–218
Recio L, Cochrane J, Simpson D, Skopek TR, O'Neill JP, Nicklas JA, Albertini RJ (1990) DNA sequence analysis of in vivo hprt mutation in human T lymphocytes. Mutagenesis 5:505–510
Rossi AM, Thijssen JCP, Tates AD, Vrieling H, Natarajan AT, Lohmann PHM, Zeeland van AA (1990) Mutations affecting RNA splicing in man are detected more frequently in somatic than in germ cells. Mutat Res 244:353–357
Salzmann J, DeMars R, Benke P (1968) Single-allele expression at an X-linked hyperuricemia locus in heterozygous human cells. Proc Natl Acad Sci USA 60:545–552
Seegmiler JE, Rosenbloom FM, Kelley WN (1967) Enzyme defect associated with a sex-linked human neurological disorder and excessive purine synthesis. Science 155:1682–1684
Skopek TU, Recio L, Simpson D, Dallaire L, Melancon SB, Ogier H, O'Neill JP, Falta MT, Nicklas JA, Albertini RJ (1990) Molecular analysis of a Lesch-Nyhan syndrome mutation (hprt Montreal) by use of T-lymphocyte cultures. Hum Genet 85:111–116
Tates AD, Bernini LF, Natarajan AT, Ploem JS, Verboerd NP, Cole J, Green MHL, Arlett CF, Norris PN (1989) Detection of somatic mutants in man: HPRT mutations in lymphocytes and hemoglobin mutations in erythrocytes. Mutat Res 213:73–82
Van Der Zee SP, Schretlen ED, Monnens LA (1968) Megaloblastic anaemia in the Lesch-Nyhan syndrome. Lancet I:1427
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Hakoda, M., Hirai, Y., Akiyama, M. et al. Selection against blood cells deficient in hypoxanthine phosphoribosyltransferase (HPRT) in Lesch-Nyhan heterozygotes occurs at the level of multipotent stem cells. Hum Genet 96, 674–680 (1995). https://doi.org/10.1007/BF00210298
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DOI: https://doi.org/10.1007/BF00210298