Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1989 Apr 1;169(4):1333–1346. doi: 10.1084/jem.169.4.1333

Mouse macrophage hemagglutinin (sheep erythrocyte receptor) with specificity for sialylated glycoconjugates characterized by a monoclonal antibody

PMCID: PMC2189241  PMID: 2926328

Abstract

An inhibitory rat mAb, SER-4, has been raised to the mouse macrophage (M phi)-restricted hemagglutinin, sheep erythrocyte receptor (SER), which binds unopsonized sheep erythrocytes through recognition of sialylated glycoconjugates. This receptor was originally defined on mouse resident bone marrow M phi where it was implicated in adhesive interactions of these cells with proliferating hematopoietic cells. In the present study using mouse serum-induced thioglycollate-elicited peritoneal M phi (TPM) as a model system for SER expression, mAb SER-4 IgG2a completely blocked rosette formation at 1 microgram/ml. The inhibition was likely to be via steric hindrance rather than through a direct interaction with the putative sialic acid binding site of SER because F(ab')2 and Fab fragments of mAb SER-4 gave a maximum inhibition of 50-60% and 0% respectively, despite binding effectively to the SER-4 antigen (Ag). Immunoprecipitation and Western blotting experiments with cultured M phi or tissue extracts demonstrated that the Ag recognized by SER-4 mAb is a single chain molecule with an apparent Mr by SDS-PAGE of 185 x 10(3) (reduced) or 170 x 10(3) (non- reduced) and is distinct from members of the leukocyte common Ag family. Expression of SER and SER-4 Ag in culture were closely correlated and depended on the presence of mouse serum for optimal induction. Further evidence that the SER-4 Ag is functionally equivalent to SER was provided by immunocytochemistry in which the overall pattern of staining in tissues was consistent with previous rosetting experiments. In the bone marrow, expression of the SER-4 Ag was restricted to the resident bone marrow M phi population with no expression on monocytes. High expression was also observed on stromal M phi within the subcapsular sinus and medullary cords in lymph nodes and on marginal metallophils in the spleen. These results therefore confirm that SER is a novel M phi-restricted receptor whose distribution and properties indicate a role in cellular interactions in hematopoietic and lymphoid tissues.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Anders E. M., Scalzo A. A., Rogers G. N., White D. O. Relationship between mitogenic activity of influenza viruses and the receptor-binding specificity of their hemagglutinin molecules. J Virol. 1986 Nov;60(2):476–482. doi: 10.1128/jvi.60.2.476-482.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Berton G., Rosen H., Ezekowitz R. A., Bellavite P., Serra M. C., Rossi F., Gordon S. Monoclonal antibodies to a particulate superoxide-forming system stimulate a respiratory burst in intact guinea pig neutrophils. Proc Natl Acad Sci U S A. 1986 Jun;83(11):4002–4006. doi: 10.1073/pnas.83.11.4002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bessis M., Mize C., Prenant M. Erythropoiesis: comparison of in vivo and in vitro amplification. Blood Cells. 1978;4(1-2):155–174. [PubMed] [Google Scholar]
  5. Buckley P. J., Smith M. R., Braverman M. F., Dickson S. A. Human spleen contains phenotypic subsets of macrophages and dendritic cells that occupy discrete microanatomic locations. Am J Pathol. 1987 Sep;128(3):505–520. [PMC free article] [PubMed] [Google Scholar]
  6. Crocker P. R., Gordon S. Isolation and characterization of resident stromal macrophages and hematopoietic cell clusters from mouse bone marrow. J Exp Med. 1985 Sep 1;162(3):993–1014. doi: 10.1084/jem.162.3.993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crocker P. R., Gordon S. Properties and distribution of a lectin-like hemagglutinin differentially expressed by murine stromal tissue macrophages. J Exp Med. 1986 Dec 1;164(6):1862–1875. doi: 10.1084/jem.164.6.1862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crocker P. R., Hill M., Gordon S. Regulation of a murine macrophage haemagglutinin (sheep erythrocyte receptor) by a species-restricted serum factor. Immunology. 1988 Dec;65(4):515–522. [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Dijkstra C. D., Van Vliet E., Döpp E. A., van der Lelij A. A., Kraal G. Marginal zone macrophages identified by a monoclonal antibody: characterization of immuno- and enzyme-histochemical properties and functional capacities. Immunology. 1985 May;55(1):23–30. [PMC free article] [PubMed] [Google Scholar]
  11. Dixon S. J., Stewart D., Grinstein S., Spiegel S. Transmembrane signaling by the B subunit of cholera toxin: increased cytoplasmic free calcium in rat lymphocytes. J Cell Biol. 1987 Sep;105(3):1153–1161. doi: 10.1083/jcb.105.3.1153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fossum S., Ford W. L. The organization of cell populations within lymph nodes: their origin, life history and functional relationships. Histopathology. 1985 May;9(5):469–499. doi: 10.1111/j.1365-2559.1985.tb02830.x. [DOI] [PubMed] [Google Scholar]
  13. Galfrè G., Milstein C., Wright B. Rat x rat hybrid myelomas and a monoclonal anti-Fd portion of mouse IgG. Nature. 1979 Jan 11;277(5692):131–133. doi: 10.1038/277131a0. [DOI] [PubMed] [Google Scholar]
  14. Haltiwanger R. S., Lehrman M. A., Eckhardt A. E., Hill R. L. The distribution and localization of the fucose-binding lectin in rat tissues and the identification of a high affinity form of the mannose/N-acetylglucosamine-binding lectin in rat liver. J Biol Chem. 1986 Jun 5;261(16):7433–7439. [PubMed] [Google Scholar]
  15. Hosein B., Bianco C. Monocyte receptors for fibronectin characterized by a monoclonal antibody that interferes with receptor activity. J Exp Med. 1985 Jul 1;162(1):157–170. doi: 10.1084/jem.162.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hume D. A., Robinson A. P., MacPherson G. G., Gordon S. The mononuclear phagocyte system of the mouse defined by immunohistochemical localization of antigen F4/80. Relationship between macrophages, Langerhans cells, reticular cells, and dendritic cells in lymphoid and hematopoietic organs. J Exp Med. 1983 Nov 1;158(5):1522–1536. doi: 10.1084/jem.158.5.1522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Humphrey J. H., Grennan D. Different macrophage populations distinguished by means of fluorescent polysaccharides. Recognition and properties of marginal-zone macrophages. Eur J Immunol. 1981 Mar;11(3):221–228. doi: 10.1002/eji.1830110311. [DOI] [PubMed] [Google Scholar]
  18. Jefferies W. A., Brandon M. R., Williams A. F., Hunt S. V. Analysis of lymphopoietic stem cells with a monoclonal antibody to the rat transferrin receptor. Immunology. 1985 Feb;54(2):333–341. [PMC free article] [PubMed] [Google Scholar]
  19. Kraal G., Janse M. Marginal metallophilic cells of the mouse spleen identified by a monoclonal antibody. Immunology. 1986 Aug;58(4):665–669. [PMC free article] [PubMed] [Google Scholar]
  20. Kraal G., Rep M., Janse M. Macrophages in T and B cell compartments and other tissue macrophages recognized by monoclonal antibody MOMA-2. An immunohistochemical study. Scand J Immunol. 1987 Dec;26(6):653–661. doi: 10.1111/j.1365-3083.1987.tb02301.x. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Lee S. H., Crocker P. R., Westaby S., Key N., Mason D. Y., Gordon S., Weatherall D. J. Isolation and immunocytochemical characterization of human bone marrow stromal macrophages in hemopoietic clusters. J Exp Med. 1988 Sep 1;168(3):1193–1198. doi: 10.1084/jem.168.3.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee S. H., Crocker P., Gordon S. Macrophage plasma membrane and secretory properties in murine malaria. Effects of Plasmodium yoelii blood-stage infection on macrophages in liver, spleen, and blood. J Exp Med. 1986 Jan 1;163(1):54–74. doi: 10.1084/jem.163.1.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lobel P., Dahms N. M., Kornfeld S. Cloning and sequence analysis of the cation-independent mannose 6-phosphate receptor. J Biol Chem. 1988 Feb 15;263(5):2563–2570. [PubMed] [Google Scholar]
  25. Matsuno K., Fujii H., Kotani M. Splenic marginal-zone macrophages and marginal metallophils in rats and mice. Cell Tissue Res. 1986;246(2):263–269. doi: 10.1007/BF00215888. [DOI] [PubMed] [Google Scholar]
  26. Nusrat A. R., Wright S. D., Aderem A. A., Steinman R. M., Cohn Z. A. Properties of isolated red pulp macrophages from mouse spleen. J Exp Med. 1988 Oct 1;168(4):1505–1510. doi: 10.1084/jem.168.4.1505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Perry V. H., Gordon S. Macrophages and microglia in the nervous system. Trends Neurosci. 1988 Jun;11(6):273–277. doi: 10.1016/0166-2236(88)90110-5. [DOI] [PubMed] [Google Scholar]
  28. Raviola E., Karnovsky M. J. Evidence for a blood-thymus barrier using electron-opaque tracers. J Exp Med. 1972 Sep 1;136(3):466–498. doi: 10.1084/jem.136.3.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Riedl M., Forster O., Rumpold H., Bernheimer H. A ganglioside-dependent cellular binding mechanism in rat macrophages. J Immunol. 1982 Mar;128(3):1205–1210. [PubMed] [Google Scholar]
  30. Roos P. H., Hartman H. J., Schlepper-Schäfer J., Kolb H., Kolb-Bachofen V. Galactose-specific receptors on liver cells. II. Characterization of the purified receptor from macrophages reveals no structural relationship to the hepatocyte receptor. Biochim Biophys Acta. 1985 Oct 30;847(1):115–121. doi: 10.1016/0167-4889(85)90161-2. [DOI] [PubMed] [Google Scholar]
  31. Rosen H., Gordon S. Monoclonal antibody to the murine type 3 complement receptor inhibits adhesion of myelomonocytic cells in vitro and inflammatory cell recruitment in vivo. J Exp Med. 1987 Dec 1;166(6):1685–1701. doi: 10.1084/jem.166.6.1685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rousseaux J., Rousseaux-Prévost R., Bazin H. Optimal conditions for the preparation of Fab and F(ab')2 fragments from monoclonal IgG of different rat IgG subclasses. J Immunol Methods. 1983 Nov 11;64(1-2):141–146. doi: 10.1016/0022-1759(83)90392-7. [DOI] [PubMed] [Google Scholar]
  33. SNOOK T. STUDIES ON THE PERIFOLLICULAR REGION OF THE RAT'S SPLEEN. Anat Rec. 1964 Feb;148:149–159. doi: 10.1002/ar.1091480205. [DOI] [PubMed] [Google Scholar]
  34. Springer T., Galfrè G., Secher D. S., Milstein C. Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Eur J Immunol. 1978 Aug;8(8):539–551. doi: 10.1002/eji.1830080802. [DOI] [PubMed] [Google Scholar]
  35. Stahl P., Schlesinger P. H., Sigardson E., Rodman J. S., Lee Y. C. Receptor-mediated pinocytosis of mannose glycoconjugates by macrophages: characterization and evidence for receptor recycling. Cell. 1980 Jan;19(1):207–215. doi: 10.1016/0092-8674(80)90402-x. [DOI] [PubMed] [Google Scholar]
  36. Thomas M. L., Lefrançois L. Differential expression of the leucocyte-common antigen family. Immunol Today. 1988 Oct;9(10):320–326. doi: 10.1016/0167-5699(88)91326-6. [DOI] [PubMed] [Google Scholar]
  37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Wileman T. E., Lennartz M. R., Stahl P. D. Identification of the macrophage mannose receptor as a 175-kDa membrane protein. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2501–2505. doi: 10.1073/pnas.83.8.2501. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES