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.
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]