Journal of Experimental Medicine, Vol 173, 1099-1109, Copyright © 1991 by Rockefeller University Press

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Cytotoxic T lymphocyte granules are secretory lysosomes, containing both perforin and granzymes

PJ Peters, J Borst, V Oorschot, M Fukuda, O Krahenbuhl, J Tschopp, JW Slot and HJ Geuze
Laboratory of Cell Biology, Medical School, University of Utrecht, The Netherlands.

Cytotoxic T lymphocytes (CTL) contain granules that are exocytosed during specific interaction with target cells (TC). In this process, the granule contents, including the lethal protein perforin, as well as granzymes, a family of serine esterases, are delivered to the TC. Information regarding the routing of these proteins towards the granule and their exact localization within the granule is of primary importance to resolve the mechanism of granule-mediated TC killing. In this study, the subcellular localization of perforin, granzymes, and known endosomal and lysosomal marker proteins was determined in human and murine CTL, by immunogold labeling of ultrathin cryosections followed by electron microscopy. Perforin and granzymes can be detected in rough endoplasmic reticulum, Golgi complex, trans-Golgi reticulum, and in all cytotoxic granules. Within the granules, they have a similar distribution and are localized not only in the so-called dense core but also over the region containing small internal vesicles. This finding implies that perforin and granzymes can be released in membrane- enveloped and/or -associated form into the intercellular cleft formed upon CTL-TC interaction. On the basis of the present evidence, additional release of these molecules in soluble form cannot be excluded. The lysosomal membrane glycoproteins lamp-1, lamp-2, and CD63, are abundantly present on the granule-delimiting outer membrane, which becomes incorporated into the CTL plasma membrane during lethal hit delivery. In contrast, the cation-dependent mannose 6-phosphate receptor, known to be present in endosomes and absent from lysosomes, is found only in a minority of the granules. Together with our previous findings that the granules are acidic and connected to the endocytic pathway, these observations define CTL granules as secretory lysosomes.
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• Hallden, G., Hadi, M., Hong, H. T., Aponte, G. W. (1999). Y Receptor-mediated Induction of CD63 Transcripts, a Tetraspanin Determined To Be Necessary for Differentiation of the Intestinal Epithelial Cell Line, hBRIE 380i Cells. J. Biol. Chem. 274: 27914-27924 [Abstract] [Full Text]  
• Martinez-Lorenzo, M. J., Anel, A., Gamen, S., Monleon, I., Lasierra, P., Larrad, L., Pineiro, A., Alava, M. A., Naval, J. (1999). Activated Human T Cells Release Bioactive Fas Ligand and APO2 Ligand in Microvesicles. J. Immunol. 163: 1274-1281 [Abstract] [Full Text]  
• Barrat, F. J., Le Deist, F., Benkerrou, M., Bousso, P., Feldmann, J., Fischer, A., de Saint Basile, G. (1999). Defective CTLA-4 cycling pathway in Chediak-Higashi syndrome: A possible mechanism for deregulation of T lymphocyte activation. Proc. Natl. Acad. Sci. USA 96: 8645-8650 [Abstract] [Full Text]  
• Andrei, C., Dazzi, C., Lotti, L., Torrisi, M. R., Chimini, G., Rubartelli, A. (1999). The Secretory Route of the Leaderless Protein Interleukin 1beta Involves Exocytosis of Endolysosome-related Vesicles. Mol. Biol. Cell 10: 1463-1475 [Abstract] [Full Text]  
• Andrejewski, N., Punnonen, E.-L., Guhde, G., Tanaka, Y., Lullmann-Rauch, R., Hartmann, D., von Figura, K., Saftig, P. (1999). Normal Lysosomal Morphology and Function in LAMP-1-deficient Mice. J. Biol. Chem. 274: 12692-12701 [Abstract] [Full Text]  
• Bird, C. H., Sutton, V. R., Sun, J., Hirst, C. E., Novak, A., Kumar, S., Trapani, J. A., Bird, P. I. (1998). Selective Regulation of Apoptosis: the Cytotoxic Lymphocyte Serpin Proteinase Inhibitor 9 Protects against Granzyme B-Mediated Apoptosis without Perturbing the Fas Cell Death Pathway. Mol. Cell. Biol. 18: 6387-6398 [Abstract] [Full Text]  
• Hammond, C., Denzin, L. K., Pan, M., Griffith, J. M., Geuze, H. J., Cresswell, P. (1998). The Tetraspan Protein CD82 Is a Resident of MHC Class II Compartments Where It Associates with HLA-DR, -DM, and -DO Molecules. J. Immunol. 161: 3282-3291 [Abstract] [Full Text]  
• Escola, J.-M., Kleijmeer, M. J., Stoorvogel, W., Griffith, J. M., Yoshie, O., Geuze, H. J. (1998). Selective Enrichment of Tetraspan Proteins on the Internal Vesicles of Multivesicular Endosomes and on Exosomes Secreted by Human B-lymphocytes. J. Biol. Chem. 273: 20121-20127 [Abstract] [Full Text]  
• Pinkoski, M. J., Hobman, M., Heibein, J. A., Tomaselli, K., Li, F., Seth, P., Froelich, C. J., Bleackley, R. C. (1998). Entry and Trafficking of Granzyme B in Target Cells During Granzyme B-Perforin-Mediated Apoptosis. Blood 92: 1044-1054 [Abstract] [Full Text]  
• Heijnen, H. F.G., Debili, N., Vainchencker, W., Breton-Gorius, J., Geuze, H. J., Sixma, J. J. (1998). Multivesicular Bodies Are an Intermediate Stage in the Formation of Platelet alpha -Granules. Blood 91: 2313-2325 [Abstract] [Full Text]  
• Molinari, M., Salio, M., Galli, C., Norais, N., Rappuoli, R., Lanzavecchia, A., Montecucco, C. (1998). Selective Inhibition of Ii-dependent Antigen Presentation by Helicobacter pylori Toxin VacA. J. Exp. Med. 187: 135-140 [Abstract] [Full Text]  
• Raposo, G., Tenza, D., Mecheri, S., Peronet, R., Bonnerot, C., Desaymard, C. (1997). Accumulation of Major Histocompatibility Complex Class II Molecules in Mast Cell Secretory Granules and Their Release upon Degranulation. Mol. Biol. Cell 8: 2631-2645 [Abstract] [Full Text]  
• Rodriguez, A., Webster, P., Ortego, J., Andrews, N. W. (1997). Lysosomes Behave as Ca2+-regulated Exocytic Vesicles in Fibroblasts and Epithelial Cells. J. Cell Biol. 137: 93-104 [Abstract] [Full Text]  
• Krenacs, L., Wellmann, A., Sorbara, L., Himmelmann, A. W., Bagdi, E., Jaffe, E. S., Raffeld, M. (1997). Cytotoxic Cell Antigen Expression in Anaplastic Large Cell Lymphomas of T- and Null-Cell Type and Hodgkin's Disease: Evidence for Distinct Cellular Origin. Blood 89: 980-989 [Abstract] [Full Text]  
• Holcombe, R. F., Baethge, B. A., Wolf, R. E., Betzing, K. W., Stewart, R. M., Hall, V. C., Fukuda, M. (1994). Correlation of Serum Interleukin-8 and Cell Surface Lysosome-associated Membrane Protein with Clinical Disease Activity in Systemic Lupus Erythematosus. Lupus 3: 97-102 [Abstract]  

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