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Carbohydrate and peptide antigens in macrophage populations derived from human bone marrow and milk: an immunomorphological and immunochemical analysis

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Summary

An immunomorphological and immunochemical study was performed to elucidate the pattern of carbohydrate antigens and their relationships to the cluster differentiation (CD) 68 epitopes on macrophages derived from human bone marrow and milk. Core and backbone antigens recognized by lectins from Bauhinia purpurea (BPA), Helix pomatia (HPA), Arachis hypogaea (PNA), Glycine max. (SBA), Griffonia simplicifolia (GSA-I-B4), Lycopersicon esculentum (LEA) and Erythrina cristagalli (ECA) were expressed by both macrophage populations. Additionally, they exhibited various peripheral type 1 and type 2 carbohydrate antigens. In bone marrow trephine biopsies, the number of macrophages stained by the CD68-specific monoclonal antibody PG-M1 exceeded significantly (range 30–40%) the subpopulation expressing SBA, GSA-I-B4 and ECA binding sites as well as the Lewisa antigen. This result is very interesting since, from in vitro studies, GSA-I-B4 and SBA are known to react especially with activated macrophages. Western blotting experiments on milk macrophage lysates revealed that ECA, GSA-I-B4, BPA, PNA and MAA visualize a 110 kDa band isographic with the CD68 antigen detected by PG-M1, KP1 and Ki-M1P monoclonal antibodies. These antibodies recognize peptide epitopes as shown by enzyme-linked immunosorbent assays after biochemical modification of milk macrophage lysates. This result is in keeping with the assumption that the CD68 antigen consists of a highly glycosylated mucin-type glycoprotein comprising various differentiation-dependent epitopes.

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References

  • balduss. e., thiele, j., charles, a., hanisch, f.-g. & fischer, r. (1994) Carbohydrate antigens of human megakaryocytes and platelet glycoproteins: a comparative study. Histochemistry 102, 205–11.

    Google Scholar 

  • budde, R., Klein, P. J. & schaefer, H. E. (1988) Lectin-, immuno- and enzymehistochemical investigations on benign and neoplastic cells of the mononuclear phagocyte system. Acta Histochemica Suppl XXXVI, 227–34.

  • dijkstra, c. d. & damoiseaux, j. g. m. c. (1993) Macrophage heterogeneity established by immunocytochemistry. Progr. Histochem. Cytochem. 27, 1–66.

    Google Scholar 

  • dorshkind, k. (1990) Regulation of hemopoiesis by bone marrow stromal cells and their products. Annu. Rev. Immunol. 8, 111–37.

    Google Scholar 

  • falini, b., flenghi, l., pileri, s., gambacorta, m., bigerna, b., durkop, h., eitelbach, f., thiele, j., pacini, r., cavaliere, a., martelli, m., cardar-elli, n., sabattini, e., poggi, s. & stein, h. (1993) PG-M1: a new monoclonal antibody directed against a fixative-resistant epitope on the macrophage-restricted form of the CD68 molecule. Am. J. Pathol. 142, 1359–72.

    Google Scholar 

  • foucar, k. & foucar, e. (1990) The mononuclear phagocyte and immunoregulatory effector (M-PIRE) system: evolving concepts. Sem. Diagn. Pathol. 7, 4–18.

    Google Scholar 

  • hanisch, f.-g., saur, a., müller, w. e. G., conrad, j. & uhlenbruck, g. (1984) Further characterization of a lectin and its in vivo receptor from Geodia cydonium. Biochim. Biophys. Acta 801, 388–95.

    Google Scholar 

  • hansmann, m.-l., kikuchi, m., wacker, h. h., radzun, h. j., nathwani, b. N., hesse, k. & parwaresch, m. r. (1992) Immunohistochemical monitoring of plasmacytoid cells in lymph node sections of Kikuchi-Fujimoto disease by a new panmacrophage antibody Ki-M1P. Hum. Pathol. 23, 676–80.

    Google Scholar 

  • holness, c. l. & simmons, d. l. (1993) Molecular cloning of CD68, a human macrophage marker related to lysosomal glycoproteins. Blood 81, 1607–13.

    Google Scholar 

  • hsu, s. m. & ree, h. j. (1983) Histochemical studies on lectin binding in reactive lymphoid tissues. J. Histochem. Cytochem. 31, 538–46.

    Google Scholar 

  • irimura, t. & nicolson, g. l. (1984) Carbohydrate chain analysis by lectin binding to electrophoretically separated glycoproteins from murine B16 melanoma sublines of various metastatic properties. Cancer Res. 44, 791–98.

    Google Scholar 

  • irimura, t., north, s. m. & nicolson, g. l. (1987) Glycoprotein profiles of macrophages at different stages of activation as revealed by lectin binding after electrophoretic separation. Eur. J. Immunol. 17, 73–8.

    Google Scholar 

  • johnston, r. b. (1988) Monocytes and macrophages. New Engl. J. Med. 318, 747–52.

    Google Scholar 

  • kreipe, h., radzun, h. j., schumacher, u. & parwaresch, m. r. (1986) Lectin binding and surface glycoprotein pattern of human macrophage populations. Histochemistry 86, 201–6.

    Google Scholar 

  • krugluger, w., gessl, a., boltz-nitulescu, g. & foerster, o. (1990) Lectin binding of rat bone marrow cells during colony-stimulating factor type 1-induced differentiation. J. Leukocyte Biol. 48, 541–8.

    Google Scholar 

  • laemmli, u. k. (1970) Cleavage of structural proteins during the assembly of the head of bacteriphage T4. Nature 227, 680–5.

    Google Scholar 

  • lee, s. h., crocker, p. r., westaby, s., key, n., mason, d. y., gordon, s. & weatherall, d. j. (1988) Isolation and immunocytochemical characterization of human bone marrow stromal macrophages in hemopoietic clusters. J. Exp. Med. 168, 1193–8.

    Google Scholar 

  • maddox, d. e., shibata, s. & goldstein, i. j. (1982) Stimulated macrophages express a new glycoprotein receptor reactive with Griffonia simplicifolia I-B4 isolectin. Proc. Natl Acad. Sci. USA 79, 166–70.

    Google Scholar 

  • petryniak, j., huard, t. k., nordblom, g. d. & goldstein, i. j. (1986) Lectin binding studies on murine peritoneal cells: physicochemical characterization of the binding of lectins from Datura stramonium, Evonymus europaea, and Griffonia simplicifolia to murine peritoneal cells. Arch. Biochem. Biophys. 244, 57–66.

    Google Scholar 

  • pulford, k. a., rigney, e. m., micklem, k. j., jones, m., stross, w. P., gatter, k. c. & mason, d. y. (1989) KP1: a new monoclonal antibody that detects a monocyte/macrophage associated antigen in routinely processed tissue sections. J. Clin. Pathol. 42, 414–21.

    Google Scholar 

  • radzun, h. j., hansmann, m.-l., heidebrecht, h. j., bödewadt-radzun, s., wacker, h. h., kreipe, h., lumbeck, h., hernandez, c., kuhn, c. & parwaresch, m. r. (1991) Detection of a monocyte/macrophage differentiation antigen in routinely processed paraffin-embedded tissues by monoclonal antibody Ki-M1P. Lab. Invest. 65, 306–15.

    Google Scholar 

  • sarker, a. b., akagi, t., yoshino, t., fujiwara, k. & murakami, i. (1993) Bauhinia purpurea lectin (BPA) binding spectra in hyperplastic human tonsil and in peripheral blood: immunohistochemical, immunoelectron microscopic, and flow cytometric analysis. J. Histochem. Cytochem. 41, 811–7.

    Google Scholar 

  • schumacheru., horny, h.-p., welsch, u. & kaiserling, e. (1991) Lectin histochemistry of human bone marrow: investigation of trephine biopsy specimens in normal and reactive states and neoplastic disorders. Histochem. J.. 23, 215–20.

    Google Scholar 

  • shimizu, y. & shaw, s. (1993) Mucins in the mainstream. Nature 366, 630–1.

    Google Scholar 

  • skansén-saphir, u., lindfors, a. & andersson, u. (1993) Cytokine production in mononuclear cells of human milk studied at the single-cell level. Pediatr. Res. 34, 213–16.

    Google Scholar 

  • speer, c. p., gahr, m. & pabst, m. j. (1986) Phagocytosis-associated oxidative metabolism in human milk macrophages. Acta Paediatr. Scand. 75, 444–51.

    Google Scholar 

  • tabor, d. r., larry, c. h. & jacobs, r. f. (1989) Differential induction of macrophage GSIB4-binding activity. J. Leukocyte Biol. 45, 452–7.

    Google Scholar 

  • thiele, j., braeckel, c., wagner, s., falini, b., dienemann, d., stein, h. & fischer, r. (1992a) Macrophages in normal human bone marrow and in chronic myeloproliferative disorders: an immunohistochemical and morphometric study by a new monoclonal antibody (PG-M1) on trephine biopsies. Virchows Arch. A Pathol. Anat. 421, 33–9.

    Google Scholar 

  • thiele, j., romatowski, c., wagner, s., dienemann, d., stein, h. & fischer, r. (1992b) Macrophages (phagocytic-histiocytic reticular cells) in reactiveinflammatory lesions of the bone marrow and in myelodysplastic syndromes (MDS). Path. Res. Pract. 188, 995–1001.

    Google Scholar 

  • titius, b. r., thiele, j., schaefer, h., kreipe, h. & fischer, r. (1994) Ki-S1 and proliferating cell nuclear antigen expression of bone marrow macrophages. Acta Haematol. 91, 144–9.

    Google Scholar 

  • towbin, h., staehelin, t. & gordon, j. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl Acad. Sci. USA 76, 4350–4.

    Google Scholar 

  • vierbuchen, m., uhlenbruck, g., ortmann, m., dufhues, g. & fischer, r. (1988) Occurrence and distribution of glycoconjugates in human tissues as detected by the Erythrina cristagalli lectin. J. Histochem. Cytochem. 36, 367–76.

    Google Scholar 

  • warfel, a. h., zucker-franklin, d. & zheng, z. y. (1991) Macrophage membrane glycoproteins that bind Griffonia simplicifolia I-B4: effect on cytotoxicity and protein secretion. J. Cell. Physiol. 147, 265–73.

    Google Scholar 

  • warfel, a. h. & zucker-franklin, d. (1992) Specific ligation of surface α-d-galactosyl epitopes markedly affects the quantity of four major proteins secreted by macrophages. J. Leukocyte Biol. 52, 80–4.

    Google Scholar 

  • woodward, m. p., young, jr., W. W. & bloodgood, r. a. (1985) Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. J. Immunol. Meth. 78, 143–53.

    Google Scholar 

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Authors and Affiliations

  1. Institute of Pathology, University of Cologne, Joseph-Stelzmann-Str. 9, 50924, Cologne, Germany

    S. E. Baldus, J. Thiele, Y. -O. Park, A. Charles, C. Mross, T. K. Zirbes, C. Wickenhauser & R. Fischer

  2. Institute of Immunobiology, University of Cologne, Kerpener Str. 15, 50924, Cologne, Germany

    F. -G. Hanisch

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  1. S. E. Baldus
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  2. J. Thiele
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  3. Y. -O. Park
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  4. A. Charles
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  5. C. Mross
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  6. F. -G. Hanisch
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  7. T. K. Zirbes
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  8. C. Wickenhauser
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  9. R. Fischer
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Baldus, S.E., Thiele, J., Park, Y.O. et al. Carbohydrate and peptide antigens in macrophage populations derived from human bone marrow and milk: an immunomorphological and immunochemical analysis. Histochem J 27, 630–638 (1995). https://doi.org/10.1007/BF00173101

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  • DOI: https://doi.org/10.1007/BF00173101

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