Summary
In this study, double labelling for major histocompatability complex (MHC) class I and class II molecules and for MHC molecules and the lysosomal membrane protein lamp-1 on ultrathin cryosections of dendritic cells isolated from human peripheral blood was performed. The plasma membrane proved to be positive for both MHC class I and MHC class II molecules and was labelled for only a very few lamp-1 molecules. MHC class I and MHC class II molecules did not co-localize intracellularly except in some peripherally located vesicles. However, many MHC class II-labelled vesicles were present in a juxtanuclear position but only some of them were co-labelled for lamp-1. These results indicate the presence of a separate, non-lysosomal compartment for class II molecules in dendritic cells.
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References
Arkema, J. M. S., Broekhuis-Fluitsma, D. M., DeLaat, P. A. J. M. & Hoefsmit, E. C. M. (1990) Human peripheral blood dendritic cells concentrate in contrast to monocytes intracellular HLA class II molecules in a juxtanuclear position. Immunobiology 181, 335–44.
Arkema, J. M. S., Schadee-Eestermans, I. L., Broekhuis-Fluitsma, D. M. & Hoefsmit, E. C. M. (1991) Localization of class II molecules in storage vesicles, endosomes and lysosomes in human dendritic cells. Immunobiology 183, 396–407.
Betjes, M. G. H., Haks, M. C., Tuk, C. W. & Beelen, R. H. J. (1991) Monoclonal antibody EBM11 (anti-CD68) discriminates between dendritic cells and macrophages after short-term culture. Immunobiology 183, 79–87.
Damoiseaux, J. G. M. C. (1991) Macrophage Heterogeneity in the Rat. Thesis, Vrije Universiteit Amsterdam, The Netherlands.
Fukuda, M., Viitala, J., Matteson, J. & Carlsson, S. R. (1988) Cloning of cDNAs encoding human lysosomal membrane glycoproteins, h-lamp-1 and h-lamp-2. Comparison of their deduced amino acid sequences. J. Biol. Chem. 263, 18 920–8.
Geuze, H. J., Slot, J. W., Scheffer, R. C. T. & Van DerLey, P. A. (1981) Use of colloidal gold particles in double-labelling immuno-electron microscopy of ultrathin frozen tissue sections. J. Cell. Biol. 89, 653–65.
Griffiths, G., Mcdowall, A., Back, R. & Dubochet, J. (1984) On the preparation of cryosections for immunochemistry. J. Ultrastruct. Res. 89, 65–78.
Kamperdijk, E. W. A., Kapsenberg, M. L., Van DenBerg, M. & Hoefsmit, E. C. M. (1985) Characterization of dendritic cells, isolated from normal and stimulated lymph nodes of the rat. Cell Tissue Res. 42, 469–74.
Knight, S. C., Farrant, J., Bryant, A., Edwards, A. J., Burman, S., Lever, A., Clarke, J. & Webster, A. D. B. (1986) Non-adherent, low-density cells from human peripheral blood contain dendritic cells and monocytes, both with veiled morphology. Immunology 57, 595–603.
Kornfeld, S. & Mellman, I. (1989) The biogenesis of lysosomes. Annu. Rev. Cell Biol. 5, 483–525.
Mane, S. M., Marzella, L., Bainton, D.F., Holt, V. K., Cha, Y., Hildreth, J. E. K. & August, J. T. (1989) Purification and characterization of human lysosomal membrane glycoproteins. Arch. Biochem. Biophys. 268, 360–78
Neefjes, J. J., Stollorz, V., Peters, P. J., Geuze, H. J. & Ploegh, H. L. (1990) The biosynthetic pathway of MHC class II but not class I molecules intersects the endocytotic route. Cell 61, 171–83.
Peters, P. J., Neefjes, J. J., Oorschot, V., Ploegh, H. L. & Geuze, H. J. (1991) MHC class II molecules segregate from MHC class I molecules in the Golgi complex for transport to lysosomal compartments. Nature 349, 669–76.
Roche, P. A. & Cresswell, P. (1990) Invariant chain association with HLA-DR molecules inhibits immunogenic peptide binding. Nature 345, 615–8.
Slot, J. W. & Geuze, H. J. (1985) A new method of preparing gold probes for multiple labelling cytochemistry. Eur. J. Cell Biol. 38, 87–93.
Tokuyasu, K. T. (1980) Immunochemistry on ultrathin frozen sections. Histochem. J. 12, 381–403.
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Arkema, J.M.S., Schadee-Eestermans, I.L., Broekhuis-Fluitsma, D.M. et al. Double labelling of major histocompatability complex molecules and lysosomal protein lamp-1 on human dendritic cells. Histochem J 26, 95–99 (1994). https://doi.org/10.1007/BF00157956
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DOI: https://doi.org/10.1007/BF00157956