Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1990 Jun;70(2):262–271.

A novel monoclonal antibody, Mar 1, directed specifically against mononuclear phagocyte system cells in rats.

A Yamashita 1, Y Hattori 1, M Kotani 1, M Miyasaka 1, T Fukumoto 1
PMCID: PMC1384204  PMID: 2197217

Abstract

Three different monoclonal antibodies (mAb), designated Mar 1, Mar 2, and Mar 3, recognizing three distinct novel antigen molecules expressed preferentially in rat macrophages, were produced by the hybridoma technique. Binding of these mAb to isolated cells or fixed cells was detected by radioactive binding assay, immunohistochemical technique and flow cytometry. Mar 1 binds specifically to the cells constituting the mononuclear phagocyte system (MPS), but not to granulocytes nor endocytosis-positive cells from non-lymphoid tissues. Mar 2 and Mar 3 recognize both the former and the latter. The isotypes of Mar 1, Mar 2 and Mar 3 were defined as IgG1, IgG1 and IgG2b, respectively. These mAb were species specific, allo-non-specific and not cytotoxic for rat peritoneal macrophages. Immunoelectron microscopic observation demonstrated that Mar 1-3 antigens are located on both surface membrane and cytoplasmic membrane structures of peritoneal macrophages, particularly on the limiting membrane of phagocytic small vesicles and large phagosomes. Immunoprecipitation experiments demonstrated that the apparent molecular weights (MW) of the reactive antigens of Mar 1, Mar 2 and Mar 3 are 95,000, 100,000 and 55,000 and 27,000, respectively. These findings indicate that all of Mar 1-3 mAb have considerable value in the identification of rat phagocytes and that, of the three kinds of antigens detected with Mar 1-3, Mar 1 antigen is a specific marker for identification of the cells constituting the MPS and may offer the means to assess the functional capability and differentiation process of the macrophage populations.

Full text

PDF
262

Images in this article

266Figure 2
on p.266
267Figure 3
on p.267
269Figure 4
on p.269
270Figure 5
on p.270

Selected References

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

  1. Austyn J. M., Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805–815. doi: 10.1002/eji.1830111013. [DOI] [PubMed] [Google Scholar]
  2. Barclay A. N. Different reticular elements in rat lymphoid tissue identified by localization of Ia, Thy-1 and MRC OX 2 antigens. Immunology. 1981 Dec;44(4):727–736. [PMC free article] [PubMed] [Google Scholar]
  3. Bodenheimer H. C., Jr, Faris R. A., Charland C., Hixson D. C. Characterization of a new monoclonal antibody to rat macrophages and Kupffer cells. Hepatology. 1988 Nov-Dec;8(6):1667–1672. doi: 10.1002/hep.1840080633. [DOI] [PubMed] [Google Scholar]
  4. Collet B., Martin A., Merdrignac G., Genetet B., Toujas L. Modulation of expression of mouse macrophage surface antigens by monoclonal antibodies. Immunology. 1985 Nov;56(3):543–548. [PMC free article] [PubMed] [Google Scholar]
  5. Dijkstra C. D., Döpp E. A., Joling P., Kraal G. The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3. Immunology. 1985 Mar;54(3):589–599. [PMC free article] [PubMed] [Google Scholar]
  6. Ishii Y., Matsuura A., Iwaki H., Takami T., Kikuchi K. Two closely related antigens expressed on granulocytes, macrophages and some reticular elements in rat lymphoid tissues: characterization by monoclonal antibodies. Immunology. 1984 Mar;51(3):477–487. [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Mason D. W., Williams A. F. The kinetics of antibody binding to membrane antigens in solution and at the cell surface. Biochem J. 1980 Apr 1;187(1):1–20. doi: 10.1042/bj1870001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mato M., Ookawara S., Aikawa E., Kawasaki K. Studies on fluorescent granular perithelium (F.G.P.) of rat cerebral cortex - especially referring to morphological changes in aging. Anat Anz. 1981;149(5):486–501. [PubMed] [Google Scholar]
  11. McMaster W. R., Williams A. F. Identification of Ia glycoproteins in rat thymus and purification from rat spleen. Eur J Immunol. 1979 Jun;9(6):426–433. doi: 10.1002/eji.1830090603. [DOI] [PubMed] [Google Scholar]
  12. Moir D. J., Ghosh A. K., Abdulaziz Z., Knight P. M., Mason D. Y. Immunoenzymatic staining of haematological samples with monoclonal antibodies. Br J Haematol. 1983 Nov;55(3):395–410. doi: 10.1111/j.1365-2141.1983.tb02154.x. [DOI] [PubMed] [Google Scholar]
  13. Morahan P. S. Macrophage nomenclature: where are we going? J Reticuloendothel Soc. 1980 Feb;27(2):223–245. [PubMed] [Google Scholar]
  14. Robinson A. P., White T. M., Mason D. W. Macrophage heterogeneity in the rat as delineated by two monoclonal antibodies MRC OX-41 and MRC OX-42, the latter recognizing complement receptor type 3. Immunology. 1986 Feb;57(2):239–247. [PMC free article] [PubMed] [Google Scholar]
  15. Rumpold H., Förster O., Böck G., Swetly P., Riedl M. Antigenic heterogeneity of rat macrophages. A monoclonal antibody reacting only with alveolar but not with other types of macrophages. Immunology. 1982 Apr;45(4):637–643. [PMC free article] [PubMed] [Google Scholar]
  16. Springer T., Galfré G., Secher D. S., Milstein C. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur J Immunol. 1979 Apr;9(4):301–306. doi: 10.1002/eji.1830090410. [DOI] [PubMed] [Google Scholar]
  17. Steinman R. M., Kaplan G., Witmer M. D., Cohn Z. A. Identification of a novel cell type in peripheral lymphoid organs of mice. V. Purification of spleen dendritic cells, new surface markers, and maintenance in vitro. J Exp Med. 1979 Jan 1;149(1):1–16. doi: 10.1084/jem.149.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Takeya M., Hsiao L., Takahashi K. A new monoclonal antibody, TRPM-3, binds specifically to certain rat macrophage populations: immunohistochemical and immunoelectron microscopic analysis. J Leukoc Biol. 1987 Mar;41(3):187–195. doi: 10.1002/jlb.41.3.187. [DOI] [PubMed] [Google Scholar]
  19. Tamura G. S., Dailey M. O., Gallatin W. M., McGrath M. S., Weissman I. L., Pillemer E. A. Isolation of molecules recognized by monoclonal antibodies and antisera: the solid phase immunoisolation technique. Anal Biochem. 1984 Feb;136(2):458–464. doi: 10.1016/0003-2697(84)90244-6. [DOI] [PubMed] [Google Scholar]
  20. Todd R. F., 3rd, Schlossman S. F. Analysis of antigenic determinants on human monocytes and macrophages. Blood. 1982 Apr;59(4):775–786. [PubMed] [Google Scholar]
  21. Wacker H. H., Radzun H. J., Parwaresch M. R. Ki-M2R, a new specific monoclonal antibody, discriminates tissue macrophages from reticulum cells and monocytes in vivo and in vitro. J Leukoc Biol. 1985 Oct;38(4):509–520. doi: 10.1002/jlb.38.4.509. [DOI] [PubMed] [Google Scholar]
  22. Witmer M. D., Steinman R. M. The anatomy of peripheral lymphoid organs with emphasis on accessory cells: light-microscopic immunocytochemical studies of mouse spleen, lymph node, and Peyer's patch. Am J Anat. 1984 Jul;170(3):465–481. doi: 10.1002/aja.1001700318. [DOI] [PubMed] [Google Scholar]
  23. Wright S. D., Rao P. E., Van Voorhis W. C., Craigmyle L. S., Iida K., Talle M. A., Westberg E. F., Goldstein G., Silverstein S. C. Identification of the C3bi receptor of human monocytes and macrophages by using monoclonal antibodies. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5699–5703. doi: 10.1073/pnas.80.18.5699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yamashita A., Miyamoto M., Hattori Y., Kimura H. Collaborative role of macrophages in interactions between parental and F1 hybrid strain lymphocytes. Transplantation. 1978 Dec;26(6):426–429. doi: 10.1097/00007890-197812000-00013. [DOI] [PubMed] [Google Scholar]

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

RESOURCES