Europe PMC

This website requires cookies, and the limited processing of your personal data in order to function. By using the site you are agreeing to this as outlined in our privacy notice and cookie policy.

Abstract 


3C10 and 1D9 are two related monoclonal antibodies that specifically identify human mononuclear phagocytes in a large number of sites, including blood monocytes, alveolar macrophages, and macrophages in tissue sections of spleen, lymph node, and skin. The antigen persists on monocytes cultured for greater than 4 wk, but it is not found on giant cells. The 3C10-1D9 determinant is carried by a 55 kD polypeptide, is expressed at approximately 40,000 copies per monocyte, and is protease sensitive. The antigen is clearly different from HLA-class II or Ia-like antigens that have been studied with a new monoclonal 9.3F10. The 9.3F10 antigen is found on B cells, dendritic cells and monocytes; is protease resistant, and occurs on a 33-29 kD doublet typical of class II products. The 3C10 monoclonal provides a clear distinction between human mononuclear phagocytes and dendritic cells. First, monocytes and lymphocytes can be eliminated from plastic-adherent mononuclear cells using 3C10, complement, and two previously described cytotoxic antibodies, BA-1 (anti-B cell) and Leu-1 (anti-T cell). As a result, the trace dendritic cell component of blood can be enriched to considerable purity (65-75%) and yield. Second, immunocytochemical staining of tissue sections reveals that 3C10+ macrophages are anatomically segregated from dendritic cells. Large numbers of 3C10+ cells are found in red pulp of spleen and in regions surrounding lymphatic channels of lymph node. However, 3C10+ macrophages are scarce in white pulp of spleen and the lymphocyte-rich cortex of node that are the sites where dendritic cells are localized. 3C10+ cells in skin are found in the dermis, particularly in leprosy infiltrates, but the Langerhans' cells of epidermis are 3C10-. The distinctive localization of macrophages and dendritic cells is consistent with their respective functions as effector and accessory cells in the immune response.

Free full text 


Logo of jexpmedLink to Publisher's site
J Exp Med. 1983 Jul 1; 158(1): 126–145.
PMCID: PMC2187086
PMID: 6190974

Specific antimononuclear phagocyte monoclonal antibodies. Application to the purification of dendritic cells and the tissue localization of macrophages

Abstract

3C10 and 1D9 are two related monoclonal antibodies that specifically identify human mononuclear phagocytes in a large number of sites, including blood monocytes, alveolar macrophages, and macrophages in tissue sections of spleen, lymph node, and skin. The antigen persists on monocytes cultured for greater than 4 wk, but it is not found on giant cells. The 3C10-1D9 determinant is carried by a 55 kD polypeptide, is expressed at approximately 40,000 copies per monocyte, and is protease sensitive. The antigen is clearly different from HLA- class II or Ia-like antigens that have been studied with a new monoclonal 9.3F10. The 9.3F10 antigen is found on B cells, dendritic cells and monocytes; is protease resistant, and occurs on a 33-29 kD doublet typical of class II products. The 3C10 monoclonal provides a clear distinction between human mononuclear phagocytes and dendritic cells. First, monocytes and lymphocytes can be eliminated from plastic- adherent mononuclear cells using 3C10, complement, and two previously described cytotoxic antibodies, BA-1 (anti-B cell) and Leu-1 (anti-T cell). As a result, the trace dendritic cell component of blood can be enriched to considerable purity (65-75%) and yield. Second, immunocytochemical staining of tissue sections reveals that 3C10+ macrophages are anatomically segregated from dendritic cells. Large numbers of 3C10+ cells are found in red pulp of spleen and in regions surrounding lymphatic channels of lymph node. However, 3C10+ macrophages are scarce in white pulp of spleen and the lymphocyte-rich cortex of node that are the sites where dendritic cells are localized. 3C10+ cells in skin are found in the dermis, particularly in leprosy infiltrates, but the Langerhans' cells of epidermis are 3C10-. The distinctive localization of macrophages and dendritic cells is consistent with their respective functions as effector and accessory cells in the immune response.

Full Text

The Full Text of this article is available as a PDF (3.1M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Austyn JM, Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805–815. [Abstract] [Google Scholar]
  • Nussenzweig MC, Steinman RM, Unkeless JC, Witmer MD, Gutchinov B, Cohn ZA. Studies of the cell surface of mouse dendritic cells and other leukocytes. J Exp Med. 1981 Jul 1;154(1):168–187. [Europe PMC free article] [Abstract] [Google Scholar]
  • Breard J, Reinherz EL, Kung PC, Goldstein G, Schlossman SF. A monoclonal antibody reactive with human peripheral blood monocytes. J Immunol. 1980 Apr;124(4):1943–1948. [Abstract] [Google Scholar]
  • Ugolini V, Nunez G, Smith RG, Stastny P, Capra JD. Initial characterization of monoclonal antibodies against human monocytes. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6764–6768. [Europe PMC free article] [Abstract] [Google Scholar]
  • Raff HV, Picker LJ, Stobo JD. Macrophage heterogeneity in man. A subpopulation of HLA-DR-bearing macrophages required for antigen-induced T cell activation also contains stimulators for autologous-reactive T cells. J Exp Med. 1980 Sep 1;152(3):581–593. [Europe PMC free article] [Abstract] [Google Scholar]
  • Ault KA, Springer TA. Cross-reaction of a rat-anti-mouse phagocyte-specific monoclonal antibody (anti-Mac-1) with human monocytes and natural killer cells. J Immunol. 1981 Jan;126(1):359–364. [Abstract] [Google Scholar]
  • Todd RF, 3rd, Nadler LM, Schlossman SF. Antigens on human monocytes identified by monoclonal antibodies. J Immunol. 1981 Apr;126(4):1435–1442. [Abstract] [Google Scholar]
  • Dimitriu-Bona A, Burmester GR, Waters SJ, Winchester RJ. Human mononuclear phagocyte differentiation antigens. I. Patterns of antigenic expression on the surface of human monocytes and macrophages defined by monoclonal antibodies. J Immunol. 1983 Jan;130(1):145–152. [Abstract] [Google Scholar]
  • Nunez G, Ugolini V, Capra JD, Stastny P. Monoclonal antibodies against human monocytes. II. Recognition of two distinct cell surface molecules. Scand J Immunol. 1982 Dec;16(6):515–523. [Abstract] [Google Scholar]
  • Todd RF, 3rd, Van Agthoven A, Schlossman SF, Terhorst C. Structural analysis of differentiation antigens Mo1 and Mo2 on human monocytes. Hybridoma. 1982;1(3):329–337. [Abstract] [Google Scholar]
  • Steinman RM, Cohn ZA. Identification of a novel cell type in peripheral lymphoid organs of mice. II. Functional properties in vitro. J Exp Med. 1974 Feb 1;139(2):380–397. [Europe PMC free article] [Abstract] [Google Scholar]
  • Steinman RM, Kaplan G, Witmer MD, Cohn ZA. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Steinman RM, Gutchinov B, Witmer MD, Nussenzweig MC. Dendritic cells are the principal stimulators of the primary mixed leukocyte reaction in mice. J Exp Med. 1983 Feb 1;157(2):613–627. [Europe PMC free article] [Abstract] [Google Scholar]
  • Van Voorhis WC, Hair LS, Steinman RM, Kaplan G. Human dendritic cells. Enrichment and characterization from peripheral blood. J Exp Med. 1982 Apr 1;155(4):1172–1187. [Europe PMC free article] [Abstract] [Google Scholar]
  • Crow MK, Kunkel HG. Human dendritic cells: major stimulators of the autologous and allogeneic mixed leucocyte reactions. Clin Exp Immunol. 1982 Aug;49(2):338–346. [Abstract] [Google Scholar]
  • Fleit HB, Wright SD, Unkeless JC. Human neutrophil Fc gamma receptor distribution and structure. Proc Natl Acad Sci U S A. 1982 May;79(10):3275–3279. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wright SD, Silverstein SC. Tumor-promoting phorbol esters stimulate C3b and C3b' receptor-mediated phagocytosis in cultured human monocytes. J Exp Med. 1982 Oct 1;156(4):1149–1164. [Europe PMC free article] [Abstract] [Google Scholar]
  • Fels AO, Pawlowski NA, Cramer EB, King TK, Cohn ZA, Scott WA. Human alveolar macrophages produce leukotriene B4. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7866–7870. [Europe PMC free article] [Abstract] [Google Scholar]
  • Stähli C, Staehelin T, Miggiano V, Schmidt J, Häring P. High frequencies of antigen-specific hybridomas: dependence on immunization parameters and prediction by spleen cell analysis. J Immunol Methods. 1980;32(3):297–304. [Abstract] [Google Scholar]
  • Unkeless JC. Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors. J Exp Med. 1979 Sep 19;150(3):580–596. [Europe PMC free article] [Abstract] [Google Scholar]
  • Omary MB, Trowbridge IS, Battifora HA. Human homologue of murine T200 glycoprotein. J Exp Med. 1980 Oct 1;152(4):842–852. [Europe PMC free article] [Abstract] [Google Scholar]
  • Reinherz EL, Kung PC, Pesando JM, Ritz J, Goldstein G, Schlossman SF. Ia determinants on human T-cell subsets defined by monoclonal antibody. Activation stimuli required for expression. J Exp Med. 1979 Dec 1;150(6):1472–1482. [Europe PMC free article] [Abstract] [Google Scholar]
  • Engleman EG, Warnke R, Fox RI, Dilley J, Benike CJ, Levy R. Studies of a human T lymphocyte antigen recognized by a monoclonal antibody. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1791–1795. [Europe PMC free article] [Abstract] [Google Scholar]
  • Fithian E, Kung P, Goldstein G, Rubenfeld M, Fenoglio C, Edelson R. Reactivity of Langerhans cells with hybridoma antibody. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2541–2544. [Europe PMC free article] [Abstract] [Google Scholar]
  • Neville DM, Jr, Glossmann H. Molecular weight determination of membrane protein and glycoprotein subunits by discontinuous gel electrophoresis in dodecyl sulfate. Methods Enzymol. 1974;32:92–102. [Abstract] [Google Scholar]
  • Fraker PJ, Speck JC., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. [Abstract] [Google Scholar]
  • 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Johnson JP, Meo T, Riethmüller G, Schendel DJ, Wank R. Direct demonstration of an HLA-DR allotypic determinant on the low molecular weight (beta) subunit using a mouse monoclonal antibody specific for DR3. J Exp Med. 1982 Jul 1;156(1):104–111. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hubbard AL, Cohn ZA. Externally disposed plasma membrane proteins. I. Enzymatic iodination of mouse L cells. J Cell Biol. 1975 Feb;64(2):438–460. [Europe PMC free article] [Abstract] [Google Scholar]
  • Li CY, Lam KW, Yam LT. Esterases in human leukocytes. J Histochem Cytochem. 1973 Jan;21(1):1–12. [Abstract] [Google Scholar]
  • Graham RC, Jr, Karnovsky MJ. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. [Abstract] [Google Scholar]
  • Van Voorhis WC, Kaplan G, Sarno EN, Horwitz MA, Steinman RM, Levis WR, Nogueira N, Hair LS, Gattass CR, Arrick BA, et al. The cutaneous infiltrates of leprosy: cellular characteristics and the predominant T-cell phenotypes. N Engl J Med. 1982 Dec 23;307(26):1593–1597. [Abstract] [Google Scholar]
  • Steinman RM, Cohn ZA. Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution. J Exp Med. 1973 May 1;137(5):1142–1162. [Europe PMC free article] [Abstract] [Google Scholar]
  • Steinman RM, Adams JC, Cohn ZA. Identification of a novel cell type in peripheral lymphoid organs of mice. IV. Identification and distribution in mouse spleen. J Exp Med. 1975 Apr 1;141(4):804–820. [Europe PMC free article] [Abstract] [Google Scholar]
  • Van Voorhis WC, Valinsky J, Hoffman E, Luban J, Hair LS, Steinman RM. Relative efficacy of human monocytes and dendritic cells as accessory cells for T cell replication. J Exp Med. 1983 Jul 1;158(1):174–191. [Europe PMC free article] [Abstract] [Google Scholar]
  • Veerman AJ. On the interdigitating cells in the thymus-dependent area of the rat spleen: a relation between the mononuclear phagocyte system and T-lymphocytes. Cell Tissue Res. 1974 Apr 11;148(2):247–257. [Abstract] [Google Scholar]
  • Eikelenboom P. Characterization of non-lymphoid cells in the white pulp of the mouse spleen: an in vivo and in vitro study. Cell Tissue Res. 1978 Dec 29;195(3):445–460. [Abstract] [Google Scholar]
  • Tew JG, Thorbecke GJ, Steinman RM. Dendritic cells in the immune response: characteristics and recommended nomenclature (A report from the Reticuloendothelial Society Committee on Nomenclature). J Reticuloendothel Soc. 1982 May;31(5):371–380. [Abstract] [Google Scholar]
  • SCOTHORNE RJ, MCGREGOR IA. Cellular changes in lymph nodes and spleen following skin homografting in the rabbit. J Anat. 1955 Jul;89(3):283–292. [Europe PMC free article] [Abstract] [Google Scholar]
  • LANGEVOORT HL. The histophysiology of the antibody response. I. Histogenesis of the plasma cell reaction in rabbit spleen. Lab Invest. 1963 Jan;12:106–118. [Abstract] [Google Scholar]
  • BURWELL RG. Studies of the primary and the secondary immune responses of lymph nodes draining homografts of fresh cancellous bone (with particular reference to mechanisms of lymph node reactivity). Ann N Y Acad Sci. 1962 Oct 24;99:821–860. [Abstract] [Google Scholar]
  • OORT J, TURK JL. A HISTOLOGICAL AND AUTORADIOGRAPHIC STUDY OF LYMPH NODES DURING THE DEVELOPMENT OF CONTACT SENSITIVITY IN THE GUINEA-PIG. Br J Exp Pathol. 1965 Apr;46:147–154. [Europe PMC free article] [Abstract] [Google Scholar]

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

Citations & impact 


Impact metrics

Jump to Citations

Citations of article over time

Alternative metrics

Altmetric item for https://www.altmetric.com/details/5338000
Altmetric
Discover the attention surrounding your research
https://www.altmetric.com/details/5338000

Article citations


Go to all (138) article citations

Protocols & materials 


Funding 


Funders who supported this work.

NCI NIH HHS (1)