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. 1983 Sep 15;214(3):711–718. doi: 10.1042/bj2140711

The metabolism of deoxyguanosine and guanosine in human B and T lymphoblasts. A role for deoxyguanosine kinase activity in the selective T-cell defect associated with purine nucleoside phosphorylase deficiency.

W R Osborne, C R Scott
PMCID: PMC1152307  PMID: 6312962

Abstract

Purine nucleoside phosphorylase (NP; EC 2.4.2.1) deficiency is associated with defective T-cell and normal B-cell immunity. Biochemical mechanisms were investigated by measuring deoxyguanosine and guanosine metabolism in normal T and B lymphoblasts and NP-deficient B lymphoblasts. Deoxyguanosine kinase activity was specifically measured by using an anti-NP antibody to prevent alternative-product formation. Kinase activity towards deoxyguanosine was significantly higher in T-cells, whereas NP activity was similar in both B- and T-cells. Only in T-cells was dGTP produced from exogenous deoxyguanosine, and this was prevented by the simultaneous addition of deoxycytidine, which resulted in a concomitant increase in GTP synthesis. Inhibition by 8-aminoguanosine of NP activity in T lymphoblasts increased formation of dGTP and decreased that of GTP from deoxyguanosine and decreased the formation of GTP from guanosine. These data suggest a central role for deoxyguanosine kinase activity in the T-cell selectivity of the immune defect.

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Selected References

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  1. Carson D. A., Kaye J., Matsumoto S., Seegmiller J. E., Thompson L. Biochemical basis for the enhanced toxicity of deoxyribonucleosides toward malignant human T cell lines. Proc Natl Acad Sci U S A. 1979 May;76(5):2430–2433. doi: 10.1073/pnas.76.5.2430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carson D. A., Kaye J., Seegmiller J. E. Lymphospecific toxicity in adenosine deaminase deficiency and purine nucleoside phosphorylase deficiency: possible role of nucleoside kinase(s). Proc Natl Acad Sci U S A. 1977 Dec;74(12):5677–5681. doi: 10.1073/pnas.74.12.5677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carson D. A., Kaye J., Wasson D. B. Differences in deoxyadenosine metabolism in human and mouse lymphocytes. J Immunol. 1980 Jan;124(1):8–12. [PubMed] [Google Scholar]
  4. Carson D. A., Kaye J., Wasson D. B. The potential importance of soluble deoxynucleotidase activity in mediating deoxyadenosine toxicity in human lymphoblasts. J Immunol. 1981 Jan;126(1):348–352. [PubMed] [Google Scholar]
  5. Cohen A., Gudas L. J., Ammann A. J., Staal G. E., Martin D. W., Jr Deoxyguanosine triphosphate as a possible toxic metabolite in the immunodeficiency associated with purine nucleoside phosphorylase deficiency. J Clin Invest. 1978 May;61(5):1405–1409. doi: 10.1172/JCI109058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen A., Lee J. W., Dosch H. M., Gelfand E. W. The expression of deoxyguanosine toxicity in T lymphocytes at different stages of maturation. J Immunol. 1980 Oct;125(4):1578–1582. [PubMed] [Google Scholar]
  7. Durham J. P., Ives D. H. Deoxycytidine kinase. II. Purification and general properties of the calf thymus enzyme. J Biol Chem. 1970 May 10;245(9):2276–2284. [PubMed] [Google Scholar]
  8. Fox R. M., Tripp E. H., Piddington S. K., Tattersall M. H. Sensitivity of leukemic human null lymphocytes to deoxynucleosides. Cancer Res. 1980 Sep;40(9):3383–3386. [PubMed] [Google Scholar]
  9. Garrett C., Santi D. V. A rapid and sensitive high pressure liquid chromatography assay for deoxyribonucleoside triphosphates in cell extracts. Anal Biochem. 1979 Nov 1;99(2):268–273. doi: 10.1016/s0003-2697(79)80005-6. [DOI] [PubMed] [Google Scholar]
  10. Gelfand E. W., Lee J. J., Dosch H. M. Selective toxicity of purine deoxynucleosides for human lymphocyte growth and function. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1998–2002. doi: 10.1073/pnas.76.4.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Giblett E. R., Ammann A. J., Wara D. W., Sandman R., Diamond L. K. Nucleoside-phosphorylase deficiency in a child with severely defective T-cell immunity and normal B-cell immunity. Lancet. 1975 May 3;1(7914):1010–1013. doi: 10.1016/s0140-6736(75)91950-9. [DOI] [PubMed] [Google Scholar]
  12. Giblett E. R., Anderson J. E., Cohen F., Pollara B., Meuwissen H. J. Adenosine-deaminase deficiency in two patients with severely impaired cellular immunity. Lancet. 1972 Nov 18;2(7786):1067–1069. doi: 10.1016/s0140-6736(72)92345-8. [DOI] [PubMed] [Google Scholar]
  13. Gower W. R., Jr, Carr M. C., Ives D. H. Deoxyguanosine kinase. Distinct molecular forms in mitochondria and cytosol. J Biol Chem. 1979 Apr 10;254(7):2180–2183. [PubMed] [Google Scholar]
  14. Gudas L. J., Ullman B., Cohen A., Martin D. W., Jr Deoxyguanosine toxicity in a mouse T lymphoma: relationship to purine nucleoside phosphorylase-associated immune dysfunction. Cell. 1978 Jul;14(3):531–538. doi: 10.1016/0092-8674(78)90239-8. [DOI] [PubMed] [Google Scholar]
  15. HOLMES R. E., ROBINS R. K. PURINE NUCLEOSIDES. IX. THE SYNTHESIS OF 9-BETA-D-RIBOFURANOSYL URIC ACID AND OTHER RELATED 8-SUBSTITUTED PURINE RIBONUCLEOSIDES. J Am Chem Soc. 1965 Apr 20;87:1772–1776. doi: 10.1021/ja01086a028. [DOI] [PubMed] [Google Scholar]
  16. Ives D. H., Durham J. P., Tucker V. S. Rapid determination of nucleoside kinase and nucleotidase activities with tritium-labeled substrates. Anal Biochem. 1969 Apr 4;28(1):192–205. doi: 10.1016/0003-2697(69)90170-5. [DOI] [PubMed] [Google Scholar]
  17. Kazmers I. S., Mitchell B. S., Dadonna P. E., Wotring L. L., Townsend L. B., Kelley W. N. Inhibition of purine nucleoside phosphorylase by 8-aminoguanosine: selective toxicity for T lymphoblasts. Science. 1981 Dec 4;214(4525):1137–1139. doi: 10.1126/science.6795718. [DOI] [PubMed] [Google Scholar]
  18. Kelley W. N., Meade J. C. Studies on hypoxanthine-guanine phosphoribosyltransferase in fibroblasts from patients with the Lesch-Nyhan syndrome. Evidence for genetic heterogeneity. J Biol Chem. 1971 May 10;246(9):2953–2958. [PubMed] [Google Scholar]
  19. Krenitsky T. A., Tuttle J. V., Koszalka G. W., Chen I. S., Beacham L. M., 3rd, Rideout J. L., Elion G. B. Deoxycytidine kinase from calf thymus. Substrate and inhibitor specificity. J Biol Chem. 1976 Jul 10;251(13):4055–4061. [PubMed] [Google Scholar]
  20. Martin D. W., Jr, Gelfand E. W. Biochemistry of diseases of immunodevelopment. Annu Rev Biochem. 1981;50:845–877. doi: 10.1146/annurev.bi.50.070181.004213. [DOI] [PubMed] [Google Scholar]
  21. Mitchell B. S., Mejias E., Daddona P. E., Kelley W. N. Purinogenic immunodeficiency diseases: selective toxicity of deoxyribonucleosides for T cells. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5011–5014. doi: 10.1073/pnas.75.10.5011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. North M. E., Newton C. A., Webster A. D. Phosphorylation of deoxyguanosine by B and T lymphocytes: evidence against selective trapping of deoxyguanosine by T lymphocytes in purine nucleoside phosphorylase deficiency. Clin Exp Immunol. 1980 Dec;42(3):523–529. [PMC free article] [PubMed] [Google Scholar]
  23. Ochs U. H., Chen S. H., Ochs H. D., Osborne W. R., Scott C. R. Purine nucleoside phosphorylase deficiency: a molecular model for selective loss of T cell function. J Immunol. 1979 Jun;122(6):2424–2429. [PubMed] [Google Scholar]
  24. Osborne W. R., Chen S. H., Giblett E. R., Biggar W. D., Ammann A. A., Scott C. R. Purine nucleoside phosphorylase deficiency. Evidence for molecular heterogeneity in two families with enzyme-deficient members. J Clin Invest. 1977 Sep;60(3):741–746. doi: 10.1172/JCI108826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Osborne W. R. Human red cell purine nucleoside phosphorylase. Purification by biospecific affinity chromatography and physical properties. J Biol Chem. 1980 Aug 10;255(15):7089–7092. [PubMed] [Google Scholar]
  26. Osborne W. R., Scott C. R. Purine nucleoside phosphorylase deficiency. Measurement of variant protein in four families with enzyme-deficient members by an enzyme-linked immunosorbent assay. Am J Hum Genet. 1980 Nov;32(6):927–933. [PMC free article] [PubMed] [Google Scholar]
  27. Osborne W. R., Sullivan J. L., Scott C. R. Formycin B, purine nucleoside phosphorylase and lymphocyte function. Immunol Commun. 1980;9(3):257–267. doi: 10.3109/08820138009065998. [DOI] [PubMed] [Google Scholar]
  28. Siegenbeek Van Heukelom L. H., Akkerman J. W., Staal G. E., De Bruyn C. H., Stoop J. W., Zegers B. J., De Bree P. K., Wadman S. K. A patient with purine nucleoside phosphorylase deficiency: enzymological and metabolic aspects. Clin Chim Acta. 1977 Feb 1;74(3):271–279. doi: 10.1016/0009-8981(77)90294-7. [DOI] [PubMed] [Google Scholar]
  29. Simmonds H. A., Sahota A., Potter C. F., Cameron J. S. Purine metabolism and immunodeficiency: urinary purine excretion as a diagnostic screening test in adenosine deaminase and purine nucleoside phosphorylase deficiency. Clin Sci Mol Med. 1978 May;54(5):579–584. doi: 10.1042/cs0540579. [DOI] [PubMed] [Google Scholar]
  30. Skupp S., Vugrek G., Ayvazian J. H. Effect of erythro-9-(2-hydroxy-3-nonyl) adenine on purine and pyrimidine metabolism in the human peripheral lymphocyte during the early phases of phytohemagglutinin-mediated blastogenesis. Biochem Pharmacol. 1979 Nov 15;28(22):3323–3331. doi: 10.1016/0006-2952(79)90128-x. [DOI] [PubMed] [Google Scholar]
  31. Stoeckler J. D., Cambor C., Kuhns V., Chu S. H., Parks R. E., Jr Inhibitors of purine nucleoside phosphorylase, C(8) and C(5') substitutions. Biochem Pharmacol. 1982 Jan 15;31(2):163–171. doi: 10.1016/0006-2952(82)90206-4. [DOI] [PubMed] [Google Scholar]
  32. Stoop J. W., Zegers B. J., Hendrickx G. F., van Heukelom L. H., Staal G. E., de Bree P. K., Wadman S. K., Ballieux R. E. Purine nucleoside phosphorylase deficiency associated with selective cellular immunodeficiency. N Engl J Med. 1977 Mar 24;296(12):651–655. doi: 10.1056/NEJM197703242961203. [DOI] [PubMed] [Google Scholar]
  33. Thelander L., Reichard P. Reduction of ribonucleotides. Annu Rev Biochem. 1979;48:133–158. doi: 10.1146/annurev.bi.48.070179.001025. [DOI] [PubMed] [Google Scholar]
  34. Ullman B., Gudas L. J., Clift S. M., Martin D. W., Jr Isolation and characterization of purine-nucleoside phosphorylase-deficient T-lymphoma cells and secondary mutants with altered ribonucleotide reductase: genetic model for immunodeficiency disease. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1074–1078. doi: 10.1073/pnas.76.3.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ullman B., Gudas L. J., Cohen A., Martin D. W., Jr Deoxyadenosine metabolism and cytotoxicity in cultured mouse T lymphoma cells: a model for immunodeficiency disease. Cell. 1978 Jun;14(2):365–375. doi: 10.1016/0092-8674(78)90122-8. [DOI] [PubMed] [Google Scholar]
  36. Ullman B., Levinson B. B., Hershfield M. S., Martin D. W., Jr A biochemical genetic study of the role of specific nucleoside kinases in deoxyadenosine phosphorylation by cultured human cells. J Biol Chem. 1981 Jan 25;256(2):848–852. [PubMed] [Google Scholar]
  37. Wentworth D. F., Wolfenden R. On the interaction of 3,4,5,6-tetrahydrouridine with human liver cytidine deaminase. Biochemistry. 1975 Nov 18;14(23):5099–5105. doi: 10.1021/bi00694a012. [DOI] [PubMed] [Google Scholar]
  38. Wilson J. M., Mitchell B. S., Daddona P. E., Kelley W. N. Purinogenic immunodeficiency diseases. Differential effects of deoxyadenosine and deoxyguanosine on DNA synthesis in human T lymphoblasts. J Clin Invest. 1979 Nov;64(5):1475–1484. doi: 10.1172/JCI109606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wortmann R. L., Mitchell B. S., Edwards N. L., Fox I. H. Biochemical basis for differential deoxyadenosine toxicity to T and B lymphoblasts: role for 5'-nucleotidase. Proc Natl Acad Sci U S A. 1979 May;76(5):2434–2437. doi: 10.1073/pnas.76.5.2434. [DOI] [PMC free article] [PubMed] [Google Scholar]

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