PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jexpmedHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
 
J Exp Med. 1988 July 1; 168(1): 157–170.
PMCID: PMC2188968

Isolation and characterization of human tonsil dendritic cells

Abstract

Human dendritic cells were isolated from tonsils by density gradient separation followed by FACS IV sorting with mAbs to remove contaminating cell populations. The resulting dendritic cell population consisted of large cells with plentiful basophilic cytoplasm, lacking in granules but containing a prominent Golgi apparatus and numerous mitochondria. The cell membrane was irregular, and marked cell protrusions were obvious when stained with anti-HLA class II reagents. Their nuclei were irregular and often indented with a visible nucleolus. These cells were not phagocytic and stimulated autologous and allogeneic lymphocytes more effectively than other tonsil cell types in MLR. Phenotypic analysis of these cells confirmed that they expressed the leucocyte common antigen and stained strongly for HLA- class II antigens. Tonsil dendritic cells also coexpressed the LFA-1 alpha and LFA-1 beta chains but did not stain with a wide variety of anti-monocyte or anti-macrophage antibodies. The cells also lacked Fc and complement receptors and failed to stain with CD1 antibodies. Extensive testing with mAbs revealed only a few positive reactions, and these were consistent with reports of these antibodies staining interdigitating cells in tissue sections. This established that tonsil dendritic cells belong to the unique haemopoietic cell lineage of dendritic cells. No cytoplasmic staining of IL-1 alpha or IL-1 beta was demonstrated, although these lymphokines were readily detected in activated monocytes.

Full Text

The Full Text of this article is available as a PDF (939K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Steinman RM, Nussenzweig MC. Dendritic cells: features and functions. Immunol Rev. 1980;53:127–147. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Inaba K, Witmer MD, Steinman RM. Clustering of dendritic cells, helper T lymphocytes, and histocompatible B cells during primary antibody responses in vitro. J Exp Med. 1984 Sep 1;160(3):858–876. [PMC free article] [PubMed]
  • Silberberg-Sinakin I, Baer RL, Thorbecke GJ. Langerhans cells: a review of their nature with emphasis on their immunologic functions. Prog Allergy. 1978;24:268–294. [PubMed]
  • Hart DN, Fabre JW. Demonstration and characterization of Ia-positive dendritic cells in the interstitial connective tissues of rat heart and other tissues, but not brain. J Exp Med. 1981 Aug 1;154(2):347–361. [PMC free article] [PubMed]
  • McKenzie JL, Beard ME, Hart DN. The effect of donor pretreatment on interstitial dendritic cell content and rat cardiac allograft survival. Transplantation. 1984 Oct;38(4):371–376. [PubMed]
  • 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. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Inaba K, Steinman RM, Van Voorhis WC, Muramatsu S. Dendritic cells are critical accessory cells for thymus-dependent antibody responses in mouse and in man. Proc Natl Acad Sci U S A. 1983 Oct;80(19):6041–6045. [PubMed]
  • Davidson SE, McKenzie JL, Beard ME, Hart DN. The tissue distribution of the 3 alpha-fucosyl-N-acetyl lactosamine determinant recognized by the CD15 monoclonal antibodies CMRF-7 and 27. Pathology. 1988 Jan;20(1):24–31. [PubMed]
  • Starling GC, Davidson SE, McKenzie JL, Hart DN. Inhibition of natural killer-cell mediated cytolysis with monoclonal antibodies to restricted and non-restricted epitopes of the leucocyte common antigen. Immunology. 1987 Jul;61(3):351–356. [PubMed]
  • McMichael AJ, Pilch JR, Galfré G, Mason DY, Fabre JW, Milstein C. A human thymocyte antigen defined by a hybrid myeloma monoclonal antibody. Eur J Immunol. 1979 Mar;9(3):205–210. [PubMed]
  • Beverley PC, Callard RE. Distinctive functional characteristics of human "T" lymphocytes defined by E rosetting or a monoclonal anti-T cell antibody. Eur J Immunol. 1981 Apr;11(4):329–334. [PubMed]
  • Vodinelich L, Tax W, Bai Y, Pegram S, Capel P, Greaves MF. A monoclonal antibody (WT1) for detecting leukemias of T-cell precursors (T-ALL). Blood. 1983 Nov;62(5):1108–1113. [PubMed]
  • Thurlow PJ, Lovering KE, McKenzie IF. A monoclonal antibody detecting a new human T cell antigen, HuLy-m2. Transplantation. 1984 Aug;38(2):143–147. [PubMed]
  • Brooks DA, Bradley J, Zola H. A differentiation antigen expressed selectively by a proportion of human blood cells: detection with a monoclonal antibody. Pathology. 1982 Jan;14(1):5–11. [PubMed]
  • Lebacq-Verheyden AM, Ravoet AM, Bazin H, Sutherland DR, Tidman N, Greaves MF. Rat AL2, AL3, AL4 and AL5 monoclonal antibodies bind to the common acute lymphoblastic leukaemia antigen (CALLA gp 100). Int J Cancer. 1983 Sep 15;32(3):273–279. [PubMed]
  • 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. [PubMed]
  • Anderson SE, McKenzie JL, McLoughlin K, Beard ME, Hart DN. The inheritance of abnormal sialoglycoproteins found in a Gerbich negative individual. Pathology. 1986 Oct;18(4):407–412. [PubMed]
  • Conlon PJ, Grabstein KH, Alpert A, Prickett KS, Hopp TP, Gillis S. Localization of human mononuclear cell interleukin 1. J Immunol. 1987 Jul 1;139(1):98–102. [PubMed]
  • Corradi MP, Jelinek DF, Ramberg JE, Lipsky PE. Development of a cell with dendritic morphology from a precursor of B lymphocyte lineage. J Immunol. 1987 Apr 1;138(7):2075–2081. [PubMed]
  • 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. [PubMed]
  • Wood GS, Turner RR, Shiurba RA, Eng L, Warnke RA. Human dendritic cells and macrophages. In situ immunophenotypic definition of subsets that exhibit specific morphologic and microenvironmental characteristics. Am J Pathol. 1985 Apr;119(1):73–82. [PubMed]
  • Hancock WW, Atkins RC. Immunohistologic analysis of the cell surface antigens of human dendritic cells using monoclonal antibodies. Transplant Proc. 1984 Aug;16(4):963–967. [PubMed]
  • Wood GS, Warner NL, Warnke RA. Anti-Leu-3/T4 antibodies react with cells of monocyte/macrophage and Langerhans lineage. J Immunol. 1983 Jul;131(1):212–216. [PubMed]
  • Schuler G, Romani N, Steinman RM. A comparison of murine epidermal Langerhans cells with spleen dendritic cells. J Invest Dermatol. 1985 Jul;85(1 Suppl):99s–106s. [PubMed]
  • Nussenzweig MC, Steinman RM, Witmer MD, Gutchinov B. A monoclonal antibody specific for mouse dendritic cells. Proc Natl Acad Sci U S A. 1982 Jan;79(1):161–165. [PubMed]
  • Poulter LW, Campbell DA, Munro C, Janossy G. Discrimination of human macrophages and dendritic cells by means of monoclonal antibodies. Scand J Immunol. 1986 Sep;24(3):351–357. [PubMed]
  • Hancock WW, Kobzik L, Colby AJ, O'Hara CJ, Cooper AG, Godleski JJ. Detection of lymphokines and lymphokine receptors in pulmonary sarcoidosis. Immunohistologic evidence that inflammatory macrophages express IL-2 receptors. Am J Pathol. 1986 Apr;123(1):1–8. [PubMed]
  • Trowbridge I. Interleukin-2 receptor proteins. Nature. 1987 Jun 11;327(6122):461–462. [PubMed]
  • Inaba K, Steinman RM. Resting and sensitized T lymphocytes exhibit distinct stimulatory (antigen-presenting cell) requirements for growth and lymphokine release. J Exp Med. 1984 Dec 1;160(6):1717–1735. [PMC free article] [PubMed]

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