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. 1985 Nov;62(2):412–420.

A B-cell hybridoma product inhibiting antibody secretion via 5'-nucleotidase.

S M Johnson
PMCID: PMC1577447  PMID: 3002687

Abstract

A mouse spleen cell/plasmacytoma fusion designed to generate hybridomas making monoclonal antibodies to human lymphocyte 5'-nucleotidase yielded three hybridomas secreting material which inhibited about 50% of human and mouse lymphocyte ecto 5'-nucleotidase activity. The inhibition proved not to be due to antibody but to material of molecular weight 44,000 +/- 7,000 which was not part of an immunoglobulin molecule, although it may be related to a B cell Fc gamma receptor. In a haemolytic plaque assay this material inhibited the production of IgG but not IgM antibody by spleen cells of mice immunized with dinitrophenylated keyhole limpit haemocyanin. By contrast, IgG production by pokeweed mitogen-stimulated human tonsillar lymphocytes (assessed by reverse haemolytic plaque assay) was partially inhibited only by ascitic fluid of one of the hybridomas. The factor was called BAN (B cell anti 5'-nucleotidase). The suppressive action of BAN on IgG production was blocked by antibodies against 5'-nucleotidase suggesting that the lymphocyte enzyme may be acting as a BAN receptor.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bailyes E. M., Newby A. C., Siddle K., Luzio J. P. Solubilization and purification of rat liver 5'-nucleotidase by use of a zwitterionic detergent and a monoclonal-antibody immunoadsorbent. Biochem J. 1982 Apr 1;203(1):245–251. doi: 10.1042/bj2030245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bailyes E. M., Soos M., Jackson P., Newby A. C., Siddle K., Luzio J. P. The existence and properties of two dimers of rat liver ecto-5'-nucleotidase. Biochem J. 1984 Jul 15;221(2):369–377. doi: 10.1042/bj2210369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brenner M. K., North M. E., Chadda H. R., Newton C. A., Malkovsky M., Webster A. D., Farrant J. The role of B cell differentiation factors and specific T cell help in the pathogenesis of primary hypogammaglobulinemia. Eur J Immunol. 1984 Nov;14(11):1021–1027. doi: 10.1002/eji.1830141111. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Cohen L., Sharp S., Kulczycki A., Jr Human monocytes, B lymphocytes, and non-B lymphocytes each have structurally unique Fc gamma receptors. J Immunol. 1983 Jul;131(1):378–383. [PubMed] [Google Scholar]
  6. Cunningham A. J., Szenberg A. Further improvements in the plaque technique for detecting single antibody-forming cells. Immunology. 1968 Apr;14(4):599–600. [PMC free article] [PubMed] [Google Scholar]
  7. Dickler H. B., Kunkel H. G. Interaction of aggregated -globulin with B lymphocytes. J Exp Med. 1972 Jul 1;136(1):191–196. doi: 10.1084/jem.136.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dickler H. B. Lymphocyte receptors for immunoglobulin. Adv Immunol. 1976;24:167–214. doi: 10.1016/s0065-2776(08)60330-2. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Evans W. H., Gurd J. W. Properties of a 5'-nucleotidase purified from mouse liver plasma membranes. Biochem J. 1973 May;133(1):189–199. doi: 10.1042/bj1330189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fridman W. H., Rabourdin-Combe C., Neauport-Sautes C., Gisler R. H. Characterization and function of T cell Fc gamma receptor. Immunol Rev. 1981;56:51–88. doi: 10.1111/j.1600-065x.1981.tb01047.x. [DOI] [PubMed] [Google Scholar]
  12. Harkness R. A., Coade S. B., Webster A. D. ATP, ADP and AMP in plasma from peripheral venous blood. Clin Chim Acta. 1984 Nov 15;143(2):91–98. doi: 10.1016/0009-8981(84)90216-x. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Massaia M., Ma D. D., Sylwestrowicz T. A., Tidman N., Price G., Janossy G., Hoffbrand A. V. Enzymes of purine metabolism in human peripheral lymphocyte subpopulations. Clin Exp Immunol. 1982 Oct;50(1):148–154. [PMC free article] [PubMed] [Google Scholar]
  17. Nilsson K., Bennich H., Johansson S. G., Pontén J. Established immunoglobulin producing myeloma (IgE) and lymphoblastoid (IgG) cell lines from an IgE myeloma patient. Clin Exp Immunol. 1970 Oct;7(4):477–489. [PMC free article] [PubMed] [Google Scholar]
  18. Pure E., Durie C. J., Summerill C. K., Unkeless J. C. Identification of soluble Fc receptors in mouse serum and the conditioned medium of stimulated B cells. J Exp Med. 1984 Dec 1;160(6):1836–1849. doi: 10.1084/jem.160.6.1836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  22. Stanley K. K., Burke B., Pitt T., Siddle K., Luzio J. P. Localisation of 5'-nucleotidase in a rat liver cell line using a monoclonal antibody and indirect immunofluorescent labelling. Exp Cell Res. 1983 Mar;144(1):39–46. doi: 10.1016/0014-4827(83)90439-1. [DOI] [PubMed] [Google Scholar]
  23. Stanley K. K., Edwards M. R., Luzio J. P. Rapid internalization of plasma-membrane 5'-nucleotidase in rat spleen lymphocytes in response to rabbit anti-(rat liver 5'-nucleotidase) serum [proceedings]. Biochem Soc Trans. 1979 Oct;7(5):1023–1024. doi: 10.1042/bst0071023. [DOI] [PubMed] [Google Scholar]
  24. Stanley K. K., Edwards M. R., Luzio J. P. Subcellular distribution and movement of 5'-nucleotidase in rat cells. Biochem J. 1980 Jan 15;186(1):59–69. doi: 10.1042/bj1860059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Takacs B. J. Characterization of functional Fc receptor material from human lymphoblastoid cell lines--I. Large scale purification and biochemical analysis. Mol Immunol. 1980 Oct;17(10):1293–1314. doi: 10.1016/0161-5890(80)90026-7. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Thompson L. F., Saxon A., O'Connor R. D., Fox R. I. Ecto-5'-nucleotidase activity in human T cell subsets. Decreased numbers of ecto-5'-nucleotidase positive cells from both OKT4+ and OKT8+ cells in patients with hypogammaglobulinemia. J Clin Invest. 1983 Apr;71(4):892–899. doi: 10.1172/JCI110843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Unkeless J. C. Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors. J Exp Med. 1979 Sep 19;150(3):580–596. doi: 10.1084/jem.150.3.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Unkeless J. C., Fleit H., Mellman I. S. Structural Aspects and Heterogeneity of Immunoglobulin Fc Receptors. Adv Immunol. 1981;31:247–270. doi: 10.1016/s0065-2776(08)60922-0. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Widnell C. C., Schneider Y. J., Pierre B., Baudhuin P., Trouet A. Evidence for a continual exchange of 5'-nucleotidase between the cell surface and cytoplasmic membranes in cultured rat fibroblasts. Cell. 1982 Jan;28(1):61–70. doi: 10.1016/0092-8674(82)90375-0. [DOI] [PubMed] [Google Scholar]
  32. de la Concha E. G., Oldham G., Webster A. D., Asherson G. L., Platts-Mills T. A. Quantitative measurements of T- and B-cell function in "variable" primary hypogammaglobulinaemia: evidence for a consistent B-cell defect. Clin Exp Immunol. 1977 Feb;27(2):208–215. [PMC free article] [PubMed] [Google Scholar]

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