Abstract
The killing mediated by cytotoxic T lymphocytes (CTL) represents an important mechanism in the immune defence against tumors and virus infections. The lytic mechanism has been proposed to consist of a polarized secretion of granule-stored molecules, occurring on effector-target cell contact. By electron microscopy, membrane deposited, pore-like lesions are detected on the target cell membrane during cytolysis by CTL. These structures resembled strikingly pores formed during complement attack.
Granules of CTL isolated by nitrogen cavitation and Percoll gradient centrifugation were shown to retain cytotoxic activity. Further purification of proteins stored in these granules led to the discovery of a membranolytic protein named perforin which was capable of polymerizing into pore-like structures. In addition to this cytolytic protein, a set of serine esterases was found as well as lysosomal enzymes and proteoglycans, whose function are not yet clearly defined. The role of perforin in the cytotoxic process is currently being explored by ablating the active gene in mice.
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References
Acha-Orbea, H., Scarpellino, L., Hertig, S., Dupuis, M., and Tschopp, J., Inhibition of lymphocyte mediated cytotoxicity by perforin antisense oligonucleotides. EMBO J.9 (1990) 3815–3819.
Berke, G., Sullivan, K. A., and Amos, D. B., Tumor immunity in vitro. Destruction of a mouse ascites tumor through a cycling pathway. Science177 (1972) 430–433.
Bhakdi, S., and Tranum Jensen, J., Membrane damage by channelforming proteins. Trends biochem. Sci.8 (1983) 134–136.
Bhakdi, S., and Tranum Jensen, J., Damage to mammalian cells by proteins that form transmembrane pores. Rev. Physiol. Biochem. Pharmac.107 (1987) 147–223.
Blumenthal, R., Millard, P. J., Henkart, M. P., Reynolds, C. W., and Henkart, P. A., Liposomes as targets for granule cytolysin from cytotoxic large granular lymphocyte tumors. Proc. natl Acad. Sci. USA81 (1984) 5551–5555.
Brunner, K. T., Mauel, J., Cerottini, J.-C., and Chapius, B., Quantitative assay of the lytic action of immune lymphoid cells on51Cr-labeled allogeneic target cells in vitro; inhibition by isoantibody and by drugs. Immunology14 (1968) 181–185.
Bykovskaja, S. N., Rytenko, A. N., Rauschenbach, M. O., and Bykovsky, A. F., Ultrastructural alteration of cytolytic T lymphocytes following their interaction with target cells. Cell Immun.40 (1978) 175–185.
Cohen, J. J., and Duke, R. C., Glucocorticoid activation of a calcium-dependent endonuclease in thymocyte nuclei leads to cell death. J. Immun.132 (1984) 38–42.
Criado, M., Lindstrom, J. M., Anderson, C. G., and Dennert, G., Cytotoxic granules from killer cells: specificity of granules and insertion of channels of defined size into target membranes. J. Immun.135 (1985) 4245–4251.
Davis, M. M., and Bjorkman, P. J., T-cell antigen receptor genes and T-cell recognition. Nature334 (1988) 395–402.
Dennert, G., and Podack, E. R., Cytolysis by H-2-specific T killer cells. Assembly of tubular complexes on target membranes. J. exp. Med.157 (1983) 1483–1495.
Dourmashkin, R., Deteix, P., Simone, C. B., and Henkart, P., Electron microscopic demonstration of lesions on target cells membranes associated with antibody dependent cellular cytotoxicity. Clin. exp. Immun.43 (1980) 554–560.
Duke, R. C., Chervenak, R., and Cohen, J. J., Endogenous endonuclease-induced DNA fragmentation: An early event in cell-mediated cytolysis. Proc. natl Acad. Sci. USA80 (1983) 6361–6365.
Erard, F., Corthesy, P., Smith, K. A., Fiers, W., Conzelmann, A., and Nabholz, M., J. exp. Med.160 (1984) 584–580
Evans, M. J., and Kaufman, M. H., Establishment in culture of pluripotent cells from mouse embryos. Nature292 (1981) 154–156.
Garcia Sanz, J. A., Velotti, F., MacDonald, H. R., Masson, D., Tschopp, J., and Nabholz, M., Appearance of granule-associated molecules during activation of cytolytic T-lymphocyte precursors by defined stimuli. Immunology64 (1988) 129–134.
Geiger, B., Rosen, D., and Berke, G., Spatial relationships of microtubule-organizing centers and the contact area of cytotoxic T-lymphocytes. J. Cell Biol.95 (1982) 137–143.
Hayes, M. P., Berrebi, G. A., and Henkart, P. A., Induction of target cell DNA release by the cytotoxic T lymphocyte granule protease granzyme A. J. exp. Med.170 (1989) 933–946.
Henkart, M. P., and Henkart, P. A., Lymphocyte mediated cytolysis as a secretory phenomenon. Adv. exp. Med. Biol.146 (1982) 227–242.
Henkart, P. A., Mechanism of lymphocyte-mediated cytotoxicity. A. Rev. Immun.3 (1985) 31–58.
Henkart, P. A., Millard, P. J., Reynolds, C. W., and Henkart, M. P., Cytolytic activity of purified cytoplasmic granules from cytotoxic rat large granular lymphocyte tumors. J. exp. Med.160 (1984) 75–93.
Henkart, P. A., and Yue, C. C., The role of cytoplasmic granules in lymphocyte cytotoxicity. Prog. Allerg.40 (1988) 82–110.
Henkart, P. A., Yue, Ch. C., Yang, J., and Rosenberg, S. A., Cytolytic and biochemical properties of cytoplasmic granules of murine lymphokine-activated killer cells. J. Immun.137 (1986) 2611–2617.
Henney, C. S., Estimation of the size of a T-cell-induced lytic lesion. Nature249 (1974) 456–450.
Ishikawa, H., Shinkai, Y., Yagita, H., Yue, C. C., Henkart, P. A., Sawada, S., Young, H. A., Reynolds, C. W. and Okumura, K., Molecular cloning of rat cytolysin. J. Immun.143 (1989) 3069–3073.
Krähenbühl, O., Rey, C., Jenne, D., Lanzavecchia, A., Groscurth, P., Carrel, S., and Tschopp, J., Characterization of granzymes A and B isolated from granules of cloned human cytotoxic T lymphocytes. J. Immun.141 (1988) 3471–3477.
Kupfer, A., and Dennert, G., Reorientation of the microtubule organizing center and the Golgi apparatus in cloned cytotoxic lymphocytes by binding to lysable target cells. J. Immun.133 (1984) 2762–2766.
Kupfer, A., Dennert, G., and Singer, S. J., The reorientation of the Golgi apparatus and the microtubule-organizing center in the cytotoxic effector is a prerequisite in the lysis of bound target cells. J. molec. cell. Immun.2 (1985) 37–49.
Kuta, A. E., Reynolds, C. R., and Henkart, P. A., Mechanism of lysis by large granular lymphocyte granule cytolysin: Generation of a stable cytolysin-RBC intermediate. J. Immun.142 (1989) 4378–4384.
Li, C. B., Gray, P. W., Lin, P. F., McGrath, K. M., Ruddle, F. H., and Ruddle, N. H., Cloning and expression of murine lymphotoxin cDNA. J. Immun.138 (1987) 4496–4501.
Lichtenheld, M. G., Olsen, K. J., Lu, P., Lowrey, D. M., Hameed, A., Hengartner, H., and Podack, E. R., Structure and function of human perforin. Nature335 (1988) 448–451.
Liu, C. C., Perussia, B., Cohn, Z. A., and Young, J. D.-E., Identification and characterization of a pore-forming protein of human peripheral blood natural killer cells. J. exp. Med.164 (1986) 2061–2076.
Liu, C. C., Steffen, M., King, F., and Young, J. D.-E. Identification, isolation, and characterization of a novel cytotoxin in murine cytolytic lymphocytes. Cell51 (1987) 393–403.
MacDonald, H. R., and Lees, R. K. Programmed death of autoreactive thymocytes. Nature343 (1990) 642–644.
Martz, E., Early steps in specific tumor cell lysis by sensitized mouse T lymphocytes. II. Electrocyte permeability increase in the target cell membrane concomitant with programming for lysis. J. Immun.117 (1976) 1023–1027.
Masson, D., Corthesy, P., Nabholz, M., and Tschopp, J., Appearance of cytolytic granules upon induction of cytolytic activity in CTL-hybrids. EMBO J.4 (1985) 2533–2538.
Masson, D., and Tschopp, J., Isolation of a lytic, pore-forming protein (perforin) from cytolytic T-lymphocytes. J. biol. Chem.260 (1985) 9069–9072.
Müller-Eberhard, H. J., Zalman, L. S., Chiu, F. J., Jung, G., and Martin, D. E., Molecular mechanisms of cytotoxicity: comparison of complement and killer lymphocytes. J. Rheumat.14 (1987) 28–34.
Olsen, I., Bou-Gharios, G., and Abraham, D., The activation of resting lymphocytes is accompanied by the biogenesis of lysosomal organelles. Eur. J. Immun.20 (1990) 2161–2170.
Pasternack, M. S., Bleier, K. J., and McInerney, T. N., Granzyme A binding to target cell proteins. Granzyme A binds to and cleaves nucleolin in vitro. J. biol. Chem.266 (1991) 14703–14708.
Peitsch, M. C., Amiguet, P., Guy, R., Brunner, J., Maizel, J. V., Jr., and Tschopp, J., Localization and molecular modelling of the membraneinserted domain of the ninth component of human complement and perforin. Molec. Immun.27 (1990) 589–602.
Peters, P. J., Geuze, H. J., van der Donk, H. A., Slot, J. W., Griffith, J. M., Stam, N. J., Clevers, H. C., and Borst, J., Molecules relevant for T cell-target cell interaction are present in cytolytic granules of human T lymphocytes. Eur. J. Immun.19 (1989) 1469–1475.
Peters, R., Sauer, H., Tschopp, J., and Fritzsch, G., Transients of perforin pore formation observed by fluorescence microscopic single channel recording. EMBO J.9 (1990) 2447–2451.
Podack, E. R., Molecular mechanisms of cytolysis by complement and by cytolytic lymphocytes. J. Cell Biochem.30 (1986) 133–170.
Podack, E. R., and Dennert, G., Assembly of two types of tubules with putative cytolytic function by cloned natural killer cells. Nature302 (1983) 442–445.
Podack, E. R., and Konigsberg, P. J., Cytolytic T cell granules. Isolation, structural biochemical, and functional characterization. J. exp. Med.160 (1984) 695–710.
Podack, E. R., and Tschopp, J., Membrane attack by complement. Molec. Immun.21 (1984) 589–603.
Podack, E. R., Young, J. D.-E., and Cohn, Z. A., Isolation and biochemical and functional characterization of perforin 1 from cytolytic T-cell granules. Proc. natl Acad. Sci. USA82 (1985) 8629–8633.
Russell, J. H., Internal disintegration model of cytotoxic lymphocyteinduced target damage. Immun. Rev.72 (1983) 97–118.
Russell, J. H., and Dobos, C. B., Mechanisms of immune lysis. II. CTL-induced nuclear disintegration of the target begins within minutes of cell contact. J. Immun.125 (1980) 1256–1261.
Russell, J. H., Masakowski, V. R., and Dobos, C. B., Mechanisms of immune lysis. I. Physiological distinction between target cell death mediated by cytotoxic T lymphocytes and antibody plus complement. J. Immun.124 (1979) 1100–1105.
Russell, J. H., Masakowski, V. R., Rucinsky, T., and Philips, G., Mechanisms of immune lysis. III. Characterization of the nature and kinetics of the cytotoxic T lymphocyte-induced nuclear lesion in the target. J. Immun.128 (1982) 2087–2094.
Shi, L., Kraut, R. P., Aebersold, R., and Greenberg, A. H. A natural killer cell granule protein that induces DNA fragmentation and apoptosis. J. exp. Med.175 (1992) 553–566.
Shinkai, Y., Takio, K., and Okumura, K., Homology of perforin to the ninth component of complement (C 9). Nature334 (1988) 525–527.
Shinkai, Y., Yoshida, M. C., Maeda, K., Kobata, T., Maruyama, K., Yodoi, J., Yagita, H., and Okumura, K., Molecular cloning and chromosomal assignment of a human perforin (PFP) gene. Immunogenetics30 (1989) 452–457.
Shiver, J. W., and Henkart, P. A., A noncytotoxic mast cell tumor line exhibits potent IgE-dependent cytotoxicity after transfection with the cytolysin/perforin gene. Cell64 (1991) 1175–1181.
Simone, C. B., and Henkart, P., Permeability changes induced in erythrocyte ghost targets by antibody-dependent cytotoxic effector cells: Evidence for membrane pores. J. Immun.124 (1980) 954–963.
Sitkovsky, M. V., Mechanistic, functional and immunopharmalogical implications of biochemical studies of antigen-receptor triggered cytolytic T-lymphocyte activation. Immun. Rev.103 (1988) 127–160.
Springer, T. A., Adhesion receptors of the immune system. Nature346 (1990) 425–434.
Stanley, K. K., Kocher, H. P., Luzio, J. P., Jackson, P., and Tschopp, J., The sequence and topology of human complement component C9. EMBO J.4 (1985) 375–382.
Sundsmo, J. S., and Müller-Eberhard, H. J., Neoantigen of the complement membrane attack complex of cytotoxic human peripheral blood lymphocytes. J. Immun.122 (1979) 2371–2378.
Thomas, K. R., and Capecchi, M. R., Site-directed mutagenesis by gene targetting in mouse embryo-derived stem cells. Cell51 (1987) 503–512.
Tian, Q., Streuli, M., Saito, H., Schlossman, S. F., and Anderson, P., A polyadenylate binding protein localized to the granules of cytolytic lymphocytes induces DNA fragmentation in target cells. Cell67 (1991) 629–639.
Tschopp, J., Masson, D., and Stanley, K. K., Structural/functional similarity between proteins involved in complement- and cytotoxic T-lymphocyte-mediated cytolysis. Nature322 (1986) 831–834.
Tschopp, J., and Nabholz, M., Perforin-mediated target cell lysis by cytolytic T lymphocytes. A. Rev. Immun.8 (1990) 279–302.
Tschopp, J., Podack, E. R., and Müller-Eberhard, H. J., Formation of transmembrane tubules by spontaneous polymerization of the hydrophilic complement protein C9. Nature, Lond.298 (1982) 534–538.
Tschopp, J., Schafer, S., Masson, D., Peitsch, M. C., and Heusser, C., Phosphorylcholine acts as a Ca2+-dependent receptor molecule for lymphocyte perforin. Nature337 (1989) 272–274.
Wright, S. C., and Bonavida, B., Selective lysis of NK sensitive target cells by a soluble mediator released from murine spleen cells and human peripheral blood lymphocytes. J. Immun.126 (1981) 1516–1521.
Wyllie, A. H., Kerr, J. F., and Currie, A. R., Cell death: the significance of apoptosis. Int. Rev. Cytol.68 (1980) 251–306.
Yannelli, J. R., Sullivan, J. A., Mandell, G. L., and Engelhard, V. H., reorientation and fusion of cytotoxic T lymphocyte granules after interaction with tarlet cells as determined by high resolution cinemicrography. J. Immun.136 (1986) 377–382.
Young, J. D.-E., Killing of target cells by lymphocytes: a mechanistic view. Physiol. Rev.69 (1989) 250–314.
Young, J. D.-E., and Cohn, Z. A., Cellular and humoral mechanisms of cytotoxicity: structural and functional analogies. Adv. Immun.41 (1987) 269–332.
Young, J. D.-E., Cohn, Z. A., and Podack, E. R., The ninth component of complement and the pore-forming protein (perforin 1) from cytotoxic T cells: structural, immunological, and functional similarities. Science233 (1986) 184–190.
Young, J. D.-E., Damiano, A., DiNome, M. A., Leong, L. G., and Cohn, Z. A., Dissociation of membrane binding and lytic activities of the lymphocyte pore-forming protein (perforin). J. exp. Med.165 (1987) 1371–1382.
Young, J. D.-E., Hengartner, H., Podack, E. R., and Cohn, Z. A., Purification and characterization of a cytolytic pore-forming protein from granules of cloned lymphocytes with natural killer activity. Cell44 (1986) 849–859.
Young, J. D.-E., Nathan, C. F., Podack, E. R., Palladino, M. A., and Cohn, Z. A., Functional channel formation associated with cytotoxic T-cell granules. Proc. natl Acad. Sci. USA83 (1986) 150–154.
Young, J. D.-E., Podack, E. R., and Cohn, Z. A. Properties of a purified pore-forming protein (perforin 1) isolated fromH-2-restricted cytotoxic T cell granules. J. exp. Med.164 (1986) 144–155.
Young, L. H., Joag, S. V., Zheng, L. M., Lee, C. P., Lee, Y. S., and Young, J. D.-E., Perforin-mediated myocardial damage in acute myocarditis. Lancet336 (1990) 1019–1021.
Zagury, D., Bernard, J., Thierness, N., Feldmann, M., and Berke, G., Isolation and characterization of individual functionally reactive cytotoxic T lymphocytes: Conjugation, killing and recycling at the single cell level. Eur. J. Immun.5 (1975) 818–810.
Zalman, L. S., Brothers, M. A., Chiu, F. J., and Müller-Eberhard, H. J., Mechanisms of cytotoxicity of human large granular lymphocytes: relationship of the cytotoxic lymphocyte protein to the ninth component (C9) of human complement. Proc. natl Acad. Sci. USA83 (1986) 5262–5266.
Zalman, L. S., Brothers, M. A., and Müller-Eberhard, H. J., A C9 related channel forming protein in the cytoplasmic granules of human large granular lymphocytes. Biosci. Rep.5 (1985) 1093–1100.
Zalman, L. S., Martin, D. E., Jung, G., and Müller-Eberhard, H. J., The cytolytic protein of human lymphocytes related to the ninth component (C9) of human complement: isolation from anti-CD 3-activated peripheral blood mononuclear cells. Proc. natl Acad. Sci. USA84 (1987) 2426–2429.
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Lowin, B., Krähenbühl, O., Müller, C. et al. Perforin and its role in T lymphocyte-mediated cytolysis. Experientia 48, 911–920 (1992). https://doi.org/10.1007/BF01919138
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DOI: https://doi.org/10.1007/BF01919138