Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1977 Jun 1;145(6):1580–1589. doi: 10.1084/jem.145.6.1580

Cultured human monocytes synthesize and secrete alpha2-macroglobulin

PMCID: PMC2180678  PMID: 68095

Abstract

alpha2-Macroglobulin levels in the supernates of cultures of different subpopulations of human peripheral blood mononuclear leukocytes were assayed by a radioimmunoassay. Unfractionated mononuclear leukocytes produced greater amounts of the macroglobulin (4.0 vs. 0.8 ng/10(6) cells) than did subpopulations enriched in T or B+T lymphocytes, by passage through nylon wool or cotton wool columns, respectively. Still higher concentrations of alpha2-macroglobulin (40 ng/10(6) cells) were measured in the supernates of glass-adherent mononuclear leukocyte cultures. These results suggest that cells of monocyte-macrophage lineage are mainly, if not exclusively, responsible for the appearance of alpha2- macroglobulin in the supernate of human peripheral blood leukocyte cultures. The de novo synthesis and release of alpha2- macroglobulin by cultured monocytes was demonstrated by immunoprecipitation of radioactivity from supernates of 32S-methionine- labeled glass-adherent cells. Antiserum against purified alpha2- macroglobulin was used in both Ouchterlony double diffusion and double antibody precipitation tests. SDS-polyacrylamide gel electrophoresis of immunoprecipitates showed that most of the radioactivity comigrated with authentic alpha2-macroglobulin subunit at about 160,000 daltons.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Barrett A. J., Starkey P. M. The interaction of alpha 2-macroglobulin with proteinases. Characteristics and specificity of the reaction, and a hypothesis concerning its molecular mechanism. Biochem J. 1973 Aug;133(4):709–724. doi: 10.1042/bj1330709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Biberfeld P., Biberfeld G., Molnar Z., Fagraeus A. Fixation of cell-bound antibody in the membrane immunofluorescence test. J Immunol Methods. 1974 Mar;4(2):135–148. doi: 10.1016/0022-1759(74)90056-8. [DOI] [PubMed] [Google Scholar]
  3. Calder E. A., Urbaniak S. J., Irvine W. J., James K. The effect of anti-alpha2-macroglobulin on K-cell cytolysis and T- and B-cell rosette formation. Clin Exp Immunol. 1975 Oct;22(1):112–120. [PMC free article] [PubMed] [Google Scholar]
  4. Cardella C. J., Davies P., Allison A. C. Immune complexes induce selective release of lysosomal hydrolases from macrophages. Nature. 1974 Jan 4;247(5435):46–48. doi: 10.1038/247046a0. [DOI] [PubMed] [Google Scholar]
  5. Debanne M. T., Bell R., Dolovich J. Uptake of proteinase-alpha-macroglobulin complexes by macrophages. Biochim Biophys Acta. 1975 Dec 5;411(2):295–304. doi: 10.1016/0304-4165(75)90309-8. [DOI] [PubMed] [Google Scholar]
  6. Gitlin D., Biasucci A. Development of gamma G, gamma A, gamma M, beta IC-beta IA, C 1 esterase inhibitor, ceruloplasmin, transferrin, hemopexin, haptoglobin, fibrinogen, plasminogen, alpha 1-antitrypsin, orosomucoid, beta-lipoprotein, alpha 2-macroglobulin, and prealbumin in the human conceptus. J Clin Invest. 1969 Aug;48(8):1433–1446. doi: 10.1172/JCI106109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gordon S., Unkeless J. C., Cohn Z. A. Induction of macrophage plasminogen activator by endotoxin stimulation and phagocytosis: evidence for a two-stage process. J Exp Med. 1974 Oct 1;140(4):995–1010. doi: 10.1084/jem.140.4.995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hovi T., Allison A. C., Williams S. C. Proliferation of human peripheral blood lymphocytes induced by A23187, a streptomyces antibiotic. Exp Cell Res. 1976 Jan;97:92–100. doi: 10.1016/0014-4827(76)90658-3. [DOI] [PubMed] [Google Scholar]
  9. Julius M. H., Simpson E., Herzenberg L. A. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973 Oct;3(10):645–649. doi: 10.1002/eji.1830031011. [DOI] [PubMed] [Google Scholar]
  10. Krohn K., Sherman L., Welch M. Studies of radioiodinated fibrinogen. I. Physicochemical properties of the ICl, chloramine-T, and electrolytic reaction products. Biochim Biophys Acta. 1972 Dec 28;285(2):404–413. doi: 10.1016/0005-2795(72)90327-3. [DOI] [PubMed] [Google Scholar]
  11. Lamvik J. O. Separation of lymphocytes from human blood. Acta Haematol. 1966 May;35(5):294–303. doi: 10.1159/000209135. [DOI] [PubMed] [Google Scholar]
  12. Mosher D. F., Wing D. A. Synthesis and secretion of alpha2-macroglobulin by cultured human fibroblasts. J Exp Med. 1976 Feb 1;143(2):462–467. doi: 10.1084/jem.143.2.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Remold H. G., Rosenberg R. D. Enhancement of migration inhibitory factor activity by plasma esterase inhibitors. J Biol Chem. 1975 Aug 25;250(16):6608–6613. [PubMed] [Google Scholar]
  14. Rice R. H., Means G. E. Radioactive labeling of proteins in vitro. J Biol Chem. 1971 Feb 10;246(3):831–832. [PubMed] [Google Scholar]
  15. SKOOG W. A., BECK W. S. Studies on the fibrinogen, dextran and phytohemagglutinin methods of isolating leukocytes. Blood. 1956 May;11(5):436–454. [PubMed] [Google Scholar]
  16. Sell S. Stimulation of rabbit peripheral lymphocytes with antisera to alpha-macroglobulin. Int Arch Allergy Appl Immunol. 1970;38(2):150–157. doi: 10.1159/000230267. [DOI] [PubMed] [Google Scholar]
  17. Stecher V. J., Thorbecke G. J. Sites of synthesis of serum proteins. OI. Serum proteins produced by macrophages in vitro. J Immunol. 1967 Oct;99(4):643–652. [PubMed] [Google Scholar]
  18. Tunstall A. M., James K. Association of alpha-macroglobulins with lymphoid cells. Nat New Biol. 1973 Nov 21;246(151):78–81. doi: 10.1038/newbio246078a0. [DOI] [PubMed] [Google Scholar]
  19. Tunstall A. M., James K. Preliminary studies on the synthesis of alpha2-macroglobulin by human lymphocytes in vitro. Clin Exp Immunol. 1974 Aug;17(4):697–701. [PMC free article] [PubMed] [Google Scholar]
  20. Tunstall A. M., Merriman J. M., Milne I., James K. Normal and pathological serum levels of alpha2-macroglobulins in men and mice. J Clin Pathol. 1975 Feb;28(2):133–139. doi: 10.1136/jcp.28.2.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vaheri A., Ruoslahti E. Fibroblast surface antigen produced but not retained by virus-transformed human cells. J Exp Med. 1975 Aug 1;142(2):530–535. doi: 10.1084/jem.142.2.530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Vassalli J. D., Hamilton J., Reich E. Macrophage plasminogen activator: modulation of enzyme production by anti-inflammatory steroids, mitotic inhibitors, and cyclic nucleotides. Cell. 1976 Jun;8(2):271–281. doi: 10.1016/0092-8674(76)90011-8. [DOI] [PubMed] [Google Scholar]
  24. WILLIAMS C. A., Jr, ASOFSKY R., THORBECKE G. J. PLASMA PROTEIN FORMATION IN VITRO BY TISSUES FROM MICE INFECTED WITH STAPHYLOCOCCI. J Exp Med. 1963 Aug 1;118:315–326. doi: 10.1084/jem.118.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Werb Z., Gordon S. Elastase secretion by stimulated macrophages. Characterization and regulation. J Exp Med. 1975 Aug 1;142(2):361–377. doi: 10.1084/jem.142.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Werb Z., Gordon S. Secretion of a specific collagenase by stimulated macrophages. J Exp Med. 1975 Aug 1;142(2):346–360. doi: 10.1084/jem.142.2.346. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES