Summary
The lysosomal compartment has been examined in activated T-lymphocytes by immunogold electron microscopy and subcellular fractionation. Immunoprecipitation and sodium dodecyl sulphate-polyacrylamide gel, electrophoresis (SDS-PAGE) of radiolabelled extracts of the T-cells showed that they contained three antigens which are fundamental to normal lysosomal function: a representative lysosomal enzyme β-glucuronidase, a lysosomal associated membrane protein (LAMP-1), and the cation-independent mannose 6-phosphate lysosomal enzyme targeting receptor (MPR). Immunogold labelling showed that β-glucuronidase was present in the rough endoplasmic reticulum, the Golgi complex and Golgi-associated vesicles. The enzyme was also found to accumulate in distinct, non-Golgi organelles in which LAMP-1 was co-localized, probably lysosomes. LAMP-1 was also found in tubular elements of the golgi and in a complex of vesicles clustered near the nucleus where MPR was also present at high density.
Fractionation of homogenates from lymphocytes on Percoll gradients revealed that β-glucuronidase was distributed throughout the low density region containing rough endoplasmic reticulum, Golgi and plasma membrane components, and the high density region which contained only lysosomal activity. Multiple immunogold electron microscopy of the latter fraction showed the presence of homogenous vesicles which had large amounts of β-glucuronidase within the lumen, LAMP-1 at the periphery and no MPR. These vesicles were probably mature lysosomes, arising from pre-lysosomal organelles enriched for LAMP-1 and MPR.
Similar content being viewed by others
References
Abraham, D. (1986) Effects of mitogenic stimulation on lymphocyte α-d-mannosidases.Eur. J. Cell Biol. 40, 167–75.
Astaldi, G., Lisiewicz, J., Cichocki, T. &Burlina, A. (1973) The lysosomal enzymes in lymphocytes. Part II. Activated lymphocytes.Acta Vitaminol. Enzymol. 27, 197–206.
Besley, G. T. N., Moss, S. E., Bain, A. D. &Dewar, A. E. (1983) Correlation of lysosomal enzyme abnormalities in various forms of adult leukaemia.J. Clin. Pathol. 36, 1000–4.
Beutler, E., Kuhl, W., Matusumoto, F. &Pangalis, G. (1976) Acid hydrolases in leukocytes and platelets of normal subjects and in patients, with Gaucher's and Fabry's disease.J. Exp. Med. 143, 975–80.
Bou-Gharios, G., Moss, J., Abraham, D., Partridge, T. &Olsen, I. (1988a) Ultrastructural studies of a lysosomal enzyme during lymphocyte activation.Br. J Exp. Pathol. 69, 661–70.
Bou-Gharios, G., Moss, J., Olsen, I. &Partridge, T. (1988b) Ultrastructural localization of a lysosomal enzyme in resin-embedded lymphocytes.Histochemistry 89, 69–74.
Brown, W. J., Goodhouse J. &Farquhar, M. G. (1986) Mannose 6-phosphate receptors for lysosomal enzymes cycle between the Golgi complex and endosomes.J. Cell Biol. 103, 1235–47.
Chen, J. W., Murphy, T. L., Willingham, M. C., Pastan, I. &August, J. T. (1985) Identification of two lysosomal membrane glycoproteins.J. Cell. Biol. 101, 85–95.
Chu, F. F. &Olden, K. (1984) Distribution of acid hydrolases in subcellular fractions of proliferating vs. non-proliferating fibroblasts.Exp. Cell Res. 154, 606–12.
Geuze, H. J., Slot, J. W., Strous, J. A., Hasilik, A. &Von Figura, K. (1985) Possible pathways for lysosomal enzyme delivery.J. Cell Biol. 101, 2253–62.
Geuze, H. J., Stoorvogel, W., Strous, G. J., Slot, J. W., Bleekemolen, J. E. &Mellman, I. (1988) Sorting of mannose 6-phosphate receptors and lysosomal membrane proteins in endocytic vesicles.J. Cell Biol. 107, 2491–501.
Green, S. A., Zimmer, K. P., Griffiths, G. &Mellman, I. (1987) Kinetics of intracellular transport and sorting of lysosomal membrane and plastic membrane proteins.J. Cell Biol. 105, 1227–40.
Griffiths, G., Hoflack, B., Simons, K., Mellman, I., &Kornfeld, S. (1988) The mannose 6-phosphate receptor and the biogenesis of lysosomes.Cell 52, 329–41.
Harms, E., Kartenbeck, J., Darai, G. &Schneider, J. (1981) Purification and characterization of human lysosomes from EB-virus transformed lymphoblasts.,Exp. Cell Res. 131, 251–66.
Haubeck, H. D., Kolsch, E., Import, M., Hasilik, A. &Von Figura, K. (1985) Natural killer cell-mediated cytotoxicity does not depend on recognition of mannose 6-phosphate residues.J. Immunol. 134, 65–9.
Hopkins, C. R., Gibson, A., Shipman, M. &Miller, K. (1990) Movement of internalized ligand-receptor complexes along a continuous endosomal reticulum.Nature 346, 335–9.
Kornfeld, S. (1987) Trafficking of lysosomal enzymes.FASEB J. 1, 462–8.
Kornfeld, S. &Mellman, I. (1989) The biogenesis of lysosomes.Annu. Rev. Cell Biol. 5, 483–525.
Lin, J., Berzofsky, J. A. &Delovitch, T. L. (1988) Ultrastructural study of internalization and recycling of antigen by antigen presenting cells.Mol. Cell Immunol. 3, 321–43.
Naperstek, Y., Cohen, I. R., Fuks, Z. &Vlodavsky, I. (1984) Activated T lymphocytes produce a matrixdegrading heparan sulphate endoglycosidase.Nature 310 241–4.
Oliver, C., Dromy, R. &Hart, T. K. (1989) Density gradient separation of two populations of lysosomes from rat parotid acinar cells.J. Histochem. Cytochem. 37, 1645–52.
Olsen, I., Bou-Gharios, G. &Abraham, D. (1990) The activation of resting lymphocytes is accompanied by the biogenesis of lysosomal organelles.Eur. J. Immunol. 20, 2161–70.
Olsen, I., Dean, M. F., Muir, H. &Harris, G. (1982) Acquisition of β-glucuronidase activity by deficient fibroblasts during direct contact with lymphoid cells.J. Cell Sci. 55, 211–31.
Pasternack, M. S. &Eisen, H. N. (1985) A novel serine esterase expressed by cytotoxic T lymphocytes.Nature 314, 743–5.
Pertoft, H., Warmegard, B. &Hook, M. (1978) Heterogeneity of lysosomes originating from rat liver parenchymal cells. Metabolic relationship of subpopulations separated by density-gradient centrifugation.Biochem. J. 174, 309–17.
Pfeffer, S. R. (1988) Mannose 6-phosphate receptors and their role in targeting proteins to lysosomes.J. Membrane Biol. 103, 7–16.
Rome, L. H., Garvin, A. J., Allietta, M. M. &Neufeld, E. F. (1979) Two species of lysosomal enzymes in the mouse pigment mutants pale ear and pallid.Mol. Cell Biochem. 31, 89–95.
Rudd, E., Kindberg, G. M., Blomhoff, H. K., Godal, T. &Berg, T. (1988) Degradation of a monoclonal anti-μ chain antibody in a human surface IgM-positive B cell line starts in prelysosomal vesicles.J. Immunol. 141, 2951–8.
Taniguchi, N., Kato, E., Yoshida, H., Iwaki, S., Ohki, T. &Koizumi, S. (1978) Alpha-glucosidase activity in human leucocytes: Choice of lymphocytes, for the diagnosis of Pompe's disease and the carrier state.Clin Chim. Acta 89, 293–9.
Telerman, A., Amson, R. B., Romasco, T., Wybran, J., Garland, P. &Mosslemans, R. (1987) Internalization of human T lymphocytes, receptors.Eur. J. Immunol. 17, 991–8.
Von Figura, K. &Hasilik, A. (1986) Lysosomal enzymes and their receptors.Annu. Rev. Biochem. 55, 167–93.
Yoshikawa, M., Watanabe, M. &Hozumi, N. (1987) Analysis of protelytic processing during specific antigen presentation.Cell. Immunol. 110, 431–5.
Zucker-Franklin, D., Grusky, G. &Yang, J.-S. (1983) Arylsulfatase in natural killer cells: Its possible role in cytotoxicity.Proc. Natl. Acad. Sci. USA 80, 6977–81.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bou-Gharios, G., Moss, J. & Olsen, I. Localization of lysosomal antigens in activated T-lymphocytes. Histochem J 23, 474–482 (1991). https://doi.org/10.1007/BF01041378
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01041378