Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1988 Mar;63(3):483–490.

Dibutyryl cyclic AMP stimulation of a monocyte-like cell line, U937: a model for monocyte chemotaxis and Fc receptor-related functions.

B Sheth 1, I Dransfield 1, L J Partridge 1, M D Barker 1, D R Burton 1
PMCID: PMC1454775  PMID: 2832314

Abstract

Treatment of the U937 cell line with 1 mM dibutyryl cyclic AMP (Bt2cAMP) resulted in a reduction in cell size and inhibition of DNA synthesis, and morphologically the cells appeared similar to macrophages. Electron micrographs indicated an increase in intracellular apparatus, whilst histochemical studies revealed smaller, denser nuclei and a greater intensity of non-specific esterase staining. Ia-like antigens (HLA-DR and HLA-DC) and complement receptor CR1 were not detected on U937 cells by monoclonal antibodies, nor were they induced by Bt2cAMP. CR3 was present in small amounts on U937 cells, and stimulation with Bt2cAMP increased the expression of this molecule in the cytoplasm and on the cell surface. Leu M3, a monocyte-specific antibody, was weakly reactive on both unstimulated and stimulated cells, whereas transferrin receptors, present on 90% of U937 cells, were lost after 48-hr stimulation with Bt2cAMP. JW6 and NH6, two monoclonal antibodies raised in our laboratory and found to be against immature monocytic antigens, showed decreased expression on stimulation. Monomer IgG binding via Fc receptors decreased on stimulated cells, and a monoclonal antibody (32.2) specific for FcRI confirmed this to be due to a decrease in the number of high-affinity receptors, rather than a decrease in IgG-binding affinity. In contrast, expression of the low-affinity FcRII, monitored by monoclonal antibody IV3, increased dramatically after stimulation. Other functional changes included the production of superoxide anions and the induction of non-specific phagocytosis. Two dimensional gel analysis, of detergent soluble proteins from unstimulated and 48-hr stimulated U937 cells, showed many differences in protein expression. A detailed investigation of these changes will facilitate a better understanding of the molecular mechanisms involved in the differentiation of U937 cells.

Full text

PDF
483

Images in this article

485Figure 2
on p.485
486Figure 3
on p.486
487Figure 6
on p.487

Selected References

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

  1. Amento E. P., Bhalla A. K., Kurnick J. T., Kradin R. L., Clemens T. L., Holick S. A., Holick M. F., Krane S. M. 1 alpha,25-dihydroxyvitamin D3 induces maturation of the human monocyte cell line U937, and, in association with a factor from human T lymphocytes, augments production of the monokine, mononuclear cell factor. J Clin Invest. 1984 Mar;73(3):731–739. doi: 10.1172/JCI111266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson C. L., Guyre P. M., Whitin J. C., Ryan D. H., Looney R. J., Fanger M. W. Monoclonal antibodies to Fc receptors for IgG on human mononuclear phagocytes. Antibody characterization and induction of superoxide production in a monocyte cell line. J Biol Chem. 1986 Sep 25;261(27):12856–12864. [PubMed] [Google Scholar]
  3. Anderson N. G., Anderson N. L. Analytical techniques for cell fractions. XXI. Two-dimensional analysis of serum and tissue proteins: multiple isoelectric focusing. Anal Biochem. 1978 Apr;85(2):331–340. doi: 10.1016/0003-2697(78)90229-4. [DOI] [PubMed] [Google Scholar]
  4. Anderson N. L., Gemmell M. A. Protein-pattern changes and morphological effects due to methionine starvation or treatment with 5-azacytidine of the phorbol-ester-sensitive cell lines HL-60, CCL-119, and U-937. Clin Chem. 1984 Dec;30(12 Pt 1):1956–1964. [PubMed] [Google Scholar]
  5. Barker M. D., Jose P. J., Williams T. J., Burton D. R. The chemoattractant des-Arg74-C5a regulates the expression of its own receptor on a monocyte-like cell line. Biochem J. 1986 Jun 1;236(2):621–624. doi: 10.1042/bj2360621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Baxter M. A., Leslie R. G., Reeves W. G. The stimulation of superoxide anion production in guinea-pig peritoneal macrophages and neutrophils by phorbol myristate acetate, opsonized zymosan and IgG2-containing soluble immune complexes. Immunology. 1983 Apr;48(4):657–665. [PMC free article] [PubMed] [Google Scholar]
  7. Cohn Z. A. Activation of mononuclear phagocytes: fact, fancy, and future. J Immunol. 1978 Sep;121(3):813–816. [PubMed] [Google Scholar]
  8. Fisher A. G., Brown G. A rapid method for determining whether monoclonal antibodies react with the same or different antigens on the cell surface. J Immunol Methods. 1980;39(4):377–385. doi: 10.1016/0022-1759(80)90238-0. [DOI] [PubMed] [Google Scholar]
  9. Gourdin M. F., Vasconcelos A. W., Tabilio A., Divine M., Farcet J. P., Reyes F. Human mononuclear phagocyte differentiation: a study of the U-937 cell line by ultrastructural cytochemistry and surface antigen analysis. Br J Haematol. 1985 Oct;61(2):281–291. doi: 10.1111/j.1365-2141.1985.tb02827.x. [DOI] [PubMed] [Google Scholar]
  10. Harris P., Ralph P. Human leukemic models of myelomonocytic development: a review of the HL-60 and U937 cell lines. J Leukoc Biol. 1985 Apr;37(4):407–422. doi: 10.1002/jlb.37.4.407. [DOI] [PubMed] [Google Scholar]
  11. Hattori T., Pack M., Bougnoux P., Chang Z. L., Hoffman T. Interferon-induced differentiation of U937 cells. Comparison with other agents that promote differentiation of human myeloid or monocytelike cell lines. J Clin Invest. 1983 Jul;72(1):237–244. doi: 10.1172/JCI110962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kaplan A. M., Brown J., Collins J. M., Morahan P. S., Snodgrass M. J. Mechanism of macrophage-mediated tumor cell cytotoxicity. J Immunol. 1978 Nov;121(5):1781–1789. [PubMed] [Google Scholar]
  13. Kay G. E., Lane B. C., Snyderman R. Induction of selective biological responses to chemoattractants in a human monocyte-like cell line. Infect Immun. 1983 Sep;41(3):1166–1174. doi: 10.1128/iai.41.3.1166-1174.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Koren H. S., Anderson S. J., Larrick J. W. In vitro activation of a human macrophage-like cell line. Nature. 1979 May 24;279(5711):328–331. doi: 10.1038/279328a0. [DOI] [PubMed] [Google Scholar]
  15. Koren H. S., Handwerger B. S., Wunderlich J. R. Identification of macrophage-like characteristics in a cultured murine tumor line. J Immunol. 1975 Feb;114(2 Pt 2):894–897. [PubMed] [Google Scholar]
  16. Kurlander R. J., Gartrell J. E. The binding and processing of monoclonal human IgG1 by cells of a human macrophage-like cell line (U937). Blood. 1983 Sep;62(3):652–662. [PubMed] [Google Scholar]
  17. Largen M. T., Tannenbaum C. S. LPS regulation of specific protein synthesis in murine peritoneal macrophages. J Immunol. 1986 Feb 1;136(3):988–993. [PubMed] [Google Scholar]
  18. Larrick J. W., Fischer D. G., Anderson S. J., Koren H. S. Characterization of a human macrophage-like cell line stimulated in vitro: a model of macrophage functions. J Immunol. 1980 Jul;125(1):6–12. [PubMed] [Google Scholar]
  19. Laskey R. A. The use of intensifying screens or organic scintillators for visualizing radioactive molecules resolved by gel electrophoresis. Methods Enzymol. 1980;65(1):363–371. doi: 10.1016/s0076-6879(80)65047-2. [DOI] [PubMed] [Google Scholar]
  20. Looney R. J., Abraham G. N., Anderson C. L. Human monocytes and U937 cells bear two distinct Fc receptors for IgG. J Immunol. 1986 Mar 1;136(5):1641–1647. [PubMed] [Google Scholar]
  21. MacKay R. J., Russell S. W. Protein changes associated with stages of activation of mouse macrophages for tumor cell killing. J Immunol. 1986 Aug 15;137(4):1392–1398. [PubMed] [Google Scholar]
  22. Minta J. O., Pambrun L. In vitro induction of cytologic and functional differentiation of the immature human monocytelike cell line U-937 with phorbol myristate acetate. Am J Pathol. 1985 Apr;119(1):111–126. [PMC free article] [PubMed] [Google Scholar]
  23. Ralph P., Harris P. E., Punjabi C. J., Welte K., Litcofsky P. B., Ho M. K., Rubin B. Y., Moore M. A., Springer T. A. Lymphokine inducing "terminal differentiation" of the human monoblast leukemia line U937: a role for gamma interferon. Blood. 1983 Dec;62(6):1169–1175. [PubMed] [Google Scholar]
  24. Schmidt A., Hattori T., Hoffman T. Differentiation of a human monocyte-like cell line by (2'-5') oligoisoadenylate. Exp Cell Res. 1984 Feb;150(2):292–297. doi: 10.1016/0014-4827(84)90571-8. [DOI] [PubMed] [Google Scholar]
  25. Steplewski Z., Lubeck M. D., Koprowski H. Human macrophages armed with murine immunoglobulin G2a antibodies to tumors destroy human cancer cells. Science. 1983 Aug 26;221(4613):865–867. doi: 10.1126/science.6879183. [DOI] [PubMed] [Google Scholar]
  26. Sundström C., Nilsson K. Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer. 1976 May 15;17(5):565–577. doi: 10.1002/ijc.2910170504. [DOI] [PubMed] [Google Scholar]
  27. Taylor I. W., Milthorpe B. K. An evaluation of DNA fluorochromes, staining techniques, and analysis for flow cytometry. I. Unperturbed cell populations. J Histochem Cytochem. 1980 Nov;28(11):1224–1232. doi: 10.1177/28.11.6159392. [DOI] [PubMed] [Google Scholar]
  28. Weiel J. E., Adams D. O., Hamilton T. A. Biochemical models of gamma-interferon action: altered expression of transferrin receptors on murine peritoneal macrophages after treatment in vitro with PMA or A23187. J Immunol. 1985 Jan;134(1):293–298. [PubMed] [Google Scholar]
  29. Woof J. M., Nik Jaafar M. I., Jefferis R., Burton D. R. The monocyte binding domain(s) on human immunoglobulin G. Mol Immunol. 1984 Jun;21(6):523–527. doi: 10.1016/0161-5890(84)90068-3. [DOI] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES