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. 1982 May;107(2):191–201.

Correlations between enzymatic and immunologic properties of human peripheral blood mononuclear cells. I. Ectoenzymes of normal and immunodeficient peripheral blood mononuclear cells.

G Losa, A Morell, S Barandun
PMCID: PMC1916004  PMID: 6123261

Abstract

The activity of plasma membrane marker enzymes which are involved in purine metabolism (5'-nucleotidase, alkaline 5'-nucleotide phosphodiesterase), in active ion transport (Na-K-Mg-adenosine triphosphatase, ouabain-sensitive Na-K-adenosine triphosphatase), in aminoacid transport (gamma-glutamyltranspeptidase), and in basic physiologic functions (alkaline phosphomonoesterase) were assayed in mononuclear cells isolated from peripheral blood of normal donors and of patients with primary immunodeficiency. Irrespective of the clinical classification of the immunodeficiency, the cells of patients were characterized by significantly diminished 5'-nucleotidase and to a certain extent by lower alkaline phosphomonoesterase activities. Average activity levels of other enzymes were similar in cells of patients and controls, but scattering was more pronounced in the first group. Determination of substrate affinity revealed different kinetic properties of 5'-nucleotidase in cells from patients and normal donors; however, the extent of inhibition by beta-glycerophosphate or alpha, beta-adenosine-methylene diphosphate was comparable for both types of cells. The presence of inhibitory compounds in patients' serum was excluded by mixing experiments. When activities of the various plasma-membrane-associated enzymes were compared with each other, significant correlations emerged in normal lymphocytes. Most of these correlations were absent in cell membranes of immunodeficient patients. The findings indicate that the plasma membrane of lymphocytes from patients with immunodeficiency may be characterized by an altered distribution of enzymatic constituents.

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Selected References

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  1. Amar-Costesec A., Wibo M., Thinès-Sempoux D., Beaufay H., Berthet J. Analytical study of microsomes and isolated subcellular membranes from rat liver. IV. Biochemical, physical, and morphological modifications of microsomal components induced by digitonin, EDTA, and pyrophosphate. J Cell Biol. 1974 Sep;62(3):717–745. doi: 10.1083/jcb.62.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersson L. C., Gahmberg C. G., Kimura A. K., Wigzell H. Activated human T lymphocytes display new surface glycoproteins. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3455–3458. doi: 10.1073/pnas.75.7.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barton R. W., Goldschneider I. 5'-Nucleotidase activity in subpopulations of rat lymphocytes. J Immunol. 1978 Dec;121(6):2329–2334. [PubMed] [Google Scholar]
  4. Beaufay H., Amar-Costesec A., Thinès-Sempoux D., Wibo M., Robbi M., Berthet J. Analytical study of microsomes and isolated subcellular membranes from rat liver. 3. Subfractionation of the microsomal fraction by isopycnic and differential centrifugation in density gradients. J Cell Biol. 1974 Apr;61(1):213–231. doi: 10.1083/jcb.61.1.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bolender R. P., Paumgartner D., Muellener D., Losa G., Weibel E. R. Integrated stereological and biochemical studies on hepatocytic membranes. I.V. Heterogeneous distribution of marker enzymes on endoplasmic reticulum membranes in fractions. J Cell Biol. 1980 Jun;85(3):577–586. doi: 10.1083/jcb.85.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  7. Carraway K. L., Doss R. C., Huggins J. W., Chesnut R. W., Carraway C. A. Effects of cytoskeletal perturbant drugs on ecto 5'-nucleotidase, a concanavalin A receptor. J Cell Biol. 1979 Dec;83(3):529–543. doi: 10.1083/jcb.83.3.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coleman M. S., Greenwood M. F., Hutton J. J., Holland P., Lampkin B., Krill C., Kastelic J. E. Adenosine deaminase, terminal deoxynucleotidyl transferase (TdT), and cell surface markers in childhood acute leukemia. Blood. 1978 Dec;52(6):1125–1131. [PubMed] [Google Scholar]
  9. Cooper M. D., Faulk W. P., Fudenberg H. H., Good R. A., Hitzig W., Kunkel H., Rosen F. S., Seligmann M., Soothill J., Wedgwood R. J. Classification of primary immunodeficiencies. N Engl J Med. 1973 May 3;288(18):966–967. doi: 10.1056/NEJM197305032881814. [DOI] [PubMed] [Google Scholar]
  10. Costantino-Ceccarini E., Novikoff P. M., Atkinson P. H., Novikoff A. B. Further characterization of HeLa S3 plasma membrane ghosts. J Cell Biol. 1978 May;77(2):448–463. doi: 10.1083/jcb.77.2.448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dornand J., Mani J. C., Mousseron-Canet M., Pau B. Propriétés d'une ATPase Ca2+ ou Mg2+ dépendante des membranes plasmiques de lymphocytes. Effet de la concanavaline A sur les ATPases membranaires. Biochimie. 1974;56(10):1425–1432. [PubMed] [Google Scholar]
  12. Dornand J., Réminiac C., Mani J. C. Activité 5'-nucléotidase des membranes plasmiques de lymphocytes. Effet de la concanavaline A. Biochimie. 1977;59(4):425–432. doi: 10.1016/s0300-9084(77)80319-2. [DOI] [PubMed] [Google Scholar]
  13. Edwards N. L., Gelfand E. W., Burk L., Dosch H. M., Fox I. H. Distribution of 5'-nucleotidase in human lymphoid tissues. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3474–3476. doi: 10.1073/pnas.76.7.3474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Edwards N. L., Magilavy D. B., Cassidy J. T., Fox I. H. Lymphocyte ecto-5'-nucleotidase deficiency in agammaglobulinemia. Science. 1978 Aug 18;201(4356):628–630. doi: 10.1126/science.27864. [DOI] [PubMed] [Google Scholar]
  15. Fishman W. H. Perspectives on alkaline phosphatase isoenzymes. Am J Med. 1974 May;56(5):617–650. doi: 10.1016/0002-9343(74)90631-7. [DOI] [PubMed] [Google Scholar]
  16. Fleit H., Conklyn M., Stebbins R. D., Silber R. Function of 5'-nucleotidase in the uptake of adenosine from AMP by human lymphocytes. J Biol Chem. 1975 Dec 10;250(23):8889–8892. [PubMed] [Google Scholar]
  17. Gahmberg C. G., Häyry P., Andersson L. C. Characterization of surface glycoproteins of mouse lymphoid cells. J Cell Biol. 1976 Mar;68(3):642–653. doi: 10.1083/jcb.68.3.642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gregory S. H., Kern M. The heterogeneous distribution of 5'-nucleotidase among rabbit lymphocytes. J Immunol. 1979 Sep;123(3):1078–1082. [PubMed] [Google Scholar]
  19. Ip S. H., Abrahm J., Cooper R. A. Enhancement of blastogenesis in cholesterol-enriched lymphocytes. J Immunol. 1980 Jan;124(1):87–93. [PubMed] [Google Scholar]
  20. Johnson S. M., Kramers M. Membrane microviscosity differences in normal and leukaemic human lymphocytes. Biochem Biophys Res Commun. 1978 Jan 30;80(2):451–457. doi: 10.1016/0006-291x(78)90698-8. [DOI] [PubMed] [Google Scholar]
  21. Johnson S. M., North M. E., Asherson G. L., Allsop J., Watts R. W., Webster A. D. Lymphocyte purine 5'-nucleotidase edficiency in primary hypogammaglobulinaemia. Lancet. 1977 Jan 22;1(8004):168–170. doi: 10.1016/s0140-6736(77)91765-2. [DOI] [PubMed] [Google Scholar]
  22. Kaplan M. E., Clark C. An improved rosetting assay for detection of human T lymphocytes. J Immunol Methods. 1974 Jul;5(2):131–135. doi: 10.1016/0022-1759(74)90003-9. [DOI] [PubMed] [Google Scholar]
  23. Lelievre L., Paraf A., Charlemagne D., Sheppard J. R. Plasma membrane studies on drug sensitive and resistant cell lines. I. Cross resistance and membrane enzyme coordination (ouabain/cAMP/Na+/K+ ATPase/adenylate cyclase). Exp Cell Res. 1977 Jan;104(1):191–197. doi: 10.1016/0014-4827(77)90081-7. [DOI] [PubMed] [Google Scholar]
  24. Meister A. Glutathione, metabolism and function via the gamma-glutamyl cycle. Life Sci. 1974 Jul 15;15(2):177–190. doi: 10.1016/0024-3205(74)90206-9. [DOI] [PubMed] [Google Scholar]
  25. Monneron A. Ultrastructural study of membrane-bound enzymes in thymocytes. J Histochem Cytochem. 1974 Dec;22(12):1128–1134. doi: 10.1177/22.12.1128. [DOI] [PubMed] [Google Scholar]
  26. Monneron A., d'Alayer J. Isolation of plasma and nuclear membranes of thymocytes. I. Enzymatic composition and ultrastructure. J Cell Biol. 1978 Apr;77(1):211–231. doi: 10.1083/jcb.77.1.211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moretta L., Ferrarini M., Mingari M. C., Moretta A., Webb S. R. Subpopulations of human T cells identified by receptors for immunoglobulins and mitogen responsiveness. J Immunol. 1976 Dec;117(6):2171–2174. [PubMed] [Google Scholar]
  28. Raz A., Collard J. G., Inbar M. Decrease in 5'-nucleotidase activity in malignant transformed and normal stimulated cells. Cancer Res. 1978 May;38(5):1258–1262. [PubMed] [Google Scholar]
  29. Rosen F. S., Janeway C. A. The gamma globulins. 3. The antibody deficiency syndromes. N Engl J Med. 1966 Sep 29;275(13):709–contd. doi: 10.1056/NEJM196609292751307. [DOI] [PubMed] [Google Scholar]
  30. Rowe M., de Gast C. G., Platts-Mills T. A., Asherson G. L., Webster A. D., Johnson S. M. 5'-nucleotidase of B and T lymphocytes isolated from human peripheral blood. Clin Exp Immunol. 1979 Apr;36(1):97–101. [PMC free article] [PubMed] [Google Scholar]
  31. Rowe M., de Gast G. C., Platts-Mills T. A., Asherson G. L., Webster A. D., Johnson S. M. Lymphocyte 5'-nucleotidase in primary hypogammaglobulinaemia and cord blood. Clin Exp Immunol. 1980 Feb;39(2):337–343. [PMC free article] [PubMed] [Google Scholar]
  32. Sachs D. H., Kiszkiss P., Kim K. J. Release of Ia antigens by a cultured B cell line. J Immunol. 1980 May;124(5):2130–2136. [PubMed] [Google Scholar]
  33. Schlager S. I., Ohanian S. H. Tumor cell lipid composition and sensitivity to humoral immune killing. II. Influence of plasma membrane and intracellular lipid and fatty acid content. J Immunol. 1980 Aug;125(2):508–517. [PubMed] [Google Scholar]
  34. Shinitzky M. An efficient method for modulation of cholesterol level in cell membranes. FEBS Lett. 1978 Jan 15;85(2):317–320. doi: 10.1016/0014-5793(78)80482-7. [DOI] [PubMed] [Google Scholar]
  35. Thompson L. F., Boss G. R., Spiegelberg H. L., Jansen I. V., O'Connor R. D., Waldmann T. A., Hamburger R. N., Seegmiller J. E. Ecto-5'-nucleotidase activity in T and B lymphocytes from normal subjects and patients with congenital X-linked agammaglobulinemia. J Immunol. 1979 Dec;123(6):2475–2478. [PubMed] [Google Scholar]
  36. Touster O., Aronson N. N., Jr, Dulaney J. T., Hendrickson H. Isolation of rat liver plasma membranes. Use of nucleotide pyrophosphatase and phosphodiesterase I as marker enzymes. J Cell Biol. 1970 Dec;47(3):604–618. doi: 10.1083/jcb.47.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wallick E. T., Lane L. K., Schwartz A. Biochemical mechanism of the sodium pump. Annu Rev Physiol. 1979;41:397–411. doi: 10.1146/annurev.ph.41.030179.002145. [DOI] [PubMed] [Google Scholar]
  38. Webster A. D., North M., Allsop J., Asherson G. L., Watts R. W. Purine metabolism in lymphocytes from patients with primary hypogammaglobulinaemia. Clin Exp Immunol. 1978 Mar;31(3):456–463. [PMC free article] [PubMed] [Google Scholar]
  39. Widnell C. C. Cytochemical localization of 5'-nucleotidase in subcellular fractions isolated from rat liver. I. The origin of 5'-nucleotidase activity in microsomes. J Cell Biol. 1972 Mar;52(3):542–558. doi: 10.1083/jcb.52.3.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wisher M. H., Evans W. H. Preparation of plasma-membrane subfractions from isolated rat hepatocytes. Biochem J. 1977 May 15;164(2):415–422. doi: 10.1042/bj1640415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. van Blitterswijk W. J., Emmelot P., Hilkmann H. A., Oomenmeulemans E. P., Inbar M. Differences in lipid fluidity among isolated plasma membranes of normal and leukemic lympocytes and membranes exfoliated from their cell surface. Biochim Biophys Acta. 1977 Jun 16;467(3):309–320. doi: 10.1016/0005-2736(77)90308-x. [DOI] [PubMed] [Google Scholar]

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