The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells

doi:10.1084/jem.176.1.47

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The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells

D Vremec, M Zorbas, R Scollay, DJ Saunders, CF Ardavin, L Wu and K Shortman
Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.

A new procedure for rapid isolation of dendritic cells (DC) was devised, involving collagenase digestion of tissues, dissociation of lymphoid-DC complexes, selection of light-density cells, then depletion of lymphocytes and other non-DC by treatment with a mixture of lineage- specific monoclonal antibodies (mAbs) and removal with anti- immunoglobulin-coupled magnetic beads. This enriched population (approximately 80% DC) was further purified when required by fluorescence-activated cell sorting for cells expressing high levels of class II major histocompatibility complex (MHC). The isolated DC were characterized by immunofluorescent staining using a panel of 30 mAbs. Thymic DC were surface positive for a number of markers characteristic of T cells, but they were distinct from T-lineage cells in expressing high levels of class II MHC, in lacking expression of the T cell receptor (TCR)-CD3 complex, and having TCR beta and gamma genes in germline state. Splenic DC shared many markers with thymic DC, but were negative for most T cell markers, with the exception of CD8. A substantial proportion of DC from both thymus and spleen expressed CD8 at high levels, comparable with that on T cells. This appeared to be authentic CD8, and was produced by the DC themselves, since they contained CD8 alpha mRNA. Thymic DC presented both the CD8 alpha and beta chains on the cell surface (Ly-2+3+), although the alpha chain was in excess; the splenic DC expressed only the CD8 alpha chain (Ly-2+3-). It is suggested that the expression of CD8 could endow certain antigen- presenting DC with a veto function.
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Mattei, F., Schiavoni, G., Belardelli, F., Tough, D. F. (2001). IL-15 Is Expressed by Dendritic Cells in Response to Type I IFN, Double-Stranded RNA, or Lipopolysaccharide and Promotes Dendritic Cell Activation. J. Immunol. 167: 1179-1187 [Abstract] [Full Text]
Zhong, W., Roberts, A. D., Woodland, D. L. (2001). Antibody-Independent Antiviral Function of Memory CD4+ T Cells In Vivo Requires Regulatory Signals from CD8+ Effector T Cells. J. Immunol. 167: 1379-1386 [Abstract] [Full Text]
Henri, S., Vremec, D., Kamath, A., Waithman, J., Williams, S., Benoist, C., Burnham, K., Saeland, S., Handman, E., Shortman, K. (2001). The Dendritic Cell Populations of Mouse Lymph Nodes. J. Immunol. 167: 741-748 [Abstract] [Full Text]
Lu, L., Bonham, C. A., Liang, X., Chen, Z., Li, W., Wang, L., Watkins, S. C., Nalesnik, M. A., Schlissel, M. S., Demestris, A. J., Fung, J. J., Qian, S. (2001). Liver-Derived DEC205+B220+CD19- Dendritic Cells Regulate T Cell Responses. J. Immunol. 166: 7042-7052 [Abstract] [Full Text]
De Smedt, T., Butz, E., Smith, J., Maldonado-Lopez, R., Pajak, B., Moser, M., Maliszewski, C. (2001). CD8{alpha}- and CD8{alpha}+ subclasses of dendritic cells undergo phenotypic and functional maturation in vitro and in vivo. J. Leukoc. Biol. 69: 951-958 [Abstract] [Full Text]
Faulkner, L., Buchan, G., Lockhart, E., Slobbe, L., Wilson, M., Baird, M. (2001). IL-2 linked to a peptide from influenza hemagglutinin enhances T cell activation by affecting the antigen-presentation function of bone marrow-derived dendritic cells. Int Immunol 13: 713-721 [Abstract] [Full Text]
Kirby, A. C., Yrlid, U., Svensson, M., Wick, M. J. (2001). Differential Involvement of Dendritic Cell Subsets During Acute Salmonella Infection. J. Immunol. 166: 6802-6811 [Abstract] [Full Text]
Manz, M. G., Traver, D., Miyamoto, T., Weissman, I. L., Akashi, K. (2001). Dendritic cell potentials of early lymphoid and myeloid progenitors. Blood 97: 3333-3341 [Abstract] [Full Text]
McIlroy, D., Troadec, C., Grassi, F., Samri, A., Barrou, B., Autran, B., Debre, P., Feuillard, J., Hosmalin, A. (2001). Investigation of human spleen dendritic cell phenotype and distribution reveals evidence of in vivo activation in a subset of organ donors. Blood 97: 3470-3477 [Abstract] [Full Text]