Abstract
Natural killer (NK) cells are CD3−, T-cell receptor (TCR)−, large granular lymphocytes (LGL) that express spontaneous lytic activity against tumor cells, virally infected cells, and perhaps certain hematopoietic progenitor cells (Trinchieri, 1989). Understanding the cellular and molecular mechanisms by which NK cells recognize and destroy target cells has become an area of considerable interest. Previous studies (Henney, 1973; Herberman et al., 1986) with NK cells have proposed to divide the process of NK killing into four identifiable stages, consisting of: (1) target cell binding (adhesion), (2) effector cell activation (recognition/ signal transduction), (3) delivery of the lethal signal to the target (lethal hit), and (4) effector cell detachment and recycling. Ca2+ plays a central role in the killing process, yet only recently has it been possible to delineate more clearly the site(s) of Ca2+ requirements in the lytic mechanism. Early studies involving cytotoxic T lymphocyte (CTL) models and more recent studies with NK cells demonstrated that Ca2+ was required at a point in the lytic process distal to Mg2+-dependent target cell adhesion but proximal to target cell disintegration (Roder and Haliotis, 1980; Quan et al., 1982; Martz et al., 1983; Berke, 1989). Recent evidence supported a role for Ca2+ in the activation of a stimulus-secretion response by killer cells. It also suggested that Ca2+ can be a potent toxic agent if allowed to accumulate at high concentrations in target cells. Therefore, Ca2+ appears to be required not only to activate killer cell function but must also enter the killer cell to activate additional processes related to stimulus-secretion coupling. Analysis of the lethal hit by which killer lymphocytes mediate target cell damage has focused on CTL and their interactions with specific target cells (Roder and Haliotis, 1980; Quan et al., 1982). It suggested, but did not prove, that during “programming for lysis” the killer cells may deposit (or secrete) materials onto the target cell that mediate the lytic signals. The hypothesis that material(s) were transferred from a killer cell to a target cell was a requisite for this target cell lysis model (Henkart and Henkart, 1982). Collectively, numerous data support this hypothesis as a general phenomenon common to numerous types of killer cells. The observation that NK cells and in vitro- activated CTL contain intracytoplasmic azurophilic granules suggested that these granules may be released during the Ca2+-dependent stage of killing and may contain materials capable of mediating target cell lysis.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Acha-Orbea H, Scarpellino L, Hertig S, Dupuis M, Tschopp J (1990): Inhibition of lymphocyte mediated cytotoxicity by perforin antisense oligonucleotides. EMBO J 9: 3815–3819
Aggarwal BB, Eessalu TE, Hass PE (1985a): Characterization of receptors for human tumor necrosis factor and regulation by y-interferon. Nature 318: 665–667.
Aggarwal BB, Henzel WJ, Moffat B, Kohr WJ, Harkins RN (1985b): Primary structure of human lymphotoxin derived from 1788 lymphoblastoid cell line. J Biol Chem 260: 2334–2344
Allbritton NL, Nagler-Anderson C, Elliott TJ, Verret CR, Eisen HN (1988a): Target cell lysis by cytotoxic T lymphocytes that lack detectable hemolytic perforin activity. J Immunol 141: 3243–3248
Allbritton NL, Verret CR, Wolley RC, Eisen HN (1988b): Calcium ion concentrations and DNA fragmentation in target cell destruction by murine cloned cytotoxic T lymphocytes. J Exp Med 167: 514–527
Anderson P, Caligiuri M, Ritz J, Schlossman SF (1989): CD3-negative natural killer cells express TCR as part of a novel molecular complex. Nature 341: 159–162
Berke G (1989): Functions and mechanisms of lysis induced by cytotoxic T lymphocytes and natural killer cells. In: Fundamental Immunology, Paul WE, ed. New York: Raven Press
Berke C, Rosen D (1987): Are lytic granules and perforin 1 involved in lysis induced by in vivo primed peritoneal exudate cytolytic T lymphocytes? Transplant Proc 19: 412–416
Berke C, Rosen D (1987): Are lytic granules and perforin 1 involved in lysis induced by in vivo primed peritoneal exudate cytolytic T lymphocytes? Transplant Proc 19: 412–416
Burkhardt JK, Hester S, Argon Y (1989): Two proteins targeted to the same lytic granule compartment undergo very different posttranslational processing. Proc Natl Acad Sci USA 86: 7128–7132
Dennert G, Podack ER (1983): Cytolysis by H-2 specific killer cells: Assembly of tubular complexes on target membranes. J Exp Med 157: 1483–1495
Duke RC, Persechini PM, Chang S, Liu C-C, Cohen JJ, Young JD-E (1989): Purified perforin induces target cell lysis but not DNA fragmentation. J Exp Med 170: 1451–1456
Engelmann H, Holtmann H, Brakebusch C, Shemer-Avni Y, Sarov I, Nophar Y, Hadas E, Leitner O, Wallach D (1990): Antibodies to a soluble form of a tumor necrosis factor (TNF) receptor have TNF-like activity. J Biol Chem 265: 14497–14504
Filippini A, Taffs RE, Agui T, Sitkovsky M (1990): EctoATPase activity in cytolytic T-lymphocytes. J Biol Chem 265: 334–340
Garcia-Sanz JA, MacDonald HR, Jenne DE, Tschopp J, Nabholz M (1990): Cell specificity of granzyme gene expression. J Immunol 145: 3111–3118
Garcia-Sanz JA, Plaetinck, G, Velotti F, Masson D, Tschopp J, MacDonald HR, Nabholz M (1987): Perforin is present only in normal activated Lyt 2+ T lymphocytes and not in L3T4+ cells, but the serine protease granzyme A is made by both subsets. EMBO J 6: 933–938
Gray PW, Aggarwal BB, Benton CV, Bringman TS, Henzel WS, Jarrett JA, Leung DW, Mofat B, Ng P, Sverdsky LP, Palladino MA, Nedwin GA (1984): Cloning and expression of cDNA for human lymphotoxin: A lymphokine with tumor necrosis activity. Nature 312: 721–724
Green LM, Reade JL, Ware CF, Devlin PE, Liang C-M, Devlin JJ (1986): Cytotoxic lymphokines produced by cloned human cytotoxic T lymphocytes. II. A novel CTL-produced cytotoxin that is antigenically distinct from tumor necrosis factor and a-lymphotoxin. J Immunol 137: 3488–3493
Gromkowski SH, Brown TC, Masson D, Tschopp J (1988): Lack of DNA degradation in target cells lysed by granules derived from cytolytic T lymphocytes. J Immunol 141: 774–778
Hameed A, Olsen KJ, Lee M-K, Lichtenheld MG, Podack ER (1989): Cytolysis by Ca-permeable transmembrane channels: Pore formation causes extensive DNA degradation and cell lysis. J Exp Med 169: 765–777
Henkart MP, Henkart PA (1982): Lymphocyte mediated cytolysis as a secretory process. In: Mechanisms in Cell Mediated Cytotoxicity, Clark and Goldstein, eds. New York: Plenum
Henkart PA, Millard PJ, Reynolds CW, Henkart MP (1984): Cytolytic activity of purified cytoplasmic granules from cytotoxic rat large granular lymphocyte tumors. J Exp Med 160: 75–93
Henney CS (1973): On the mechanism of T-cell mediated cytolysis. Transplant Rev 17: 37–41
Herberman RB, Reynolds CW, Ortaldo JR (1986): Mechanism of cytotoxicity by natural killer (NK) cells. Annu Rev Immunol 4: 651–680
Hommel-Berrey G, Goebel WS, Bajpai A, Shenoy AM, Brahmi Z (1990): Loss of serine esterase activity and granule protein messages from CTL inactivated by a sensitive target. 4th International Cell-Mediated Cytotoxicity Workshop, Ogelbay Park, WV (Abstract )
Inverardi L, Witson JC, Fuad SA, Winkler-Pickett RT, Ortaldo JR, Bach FH (1991): CD3 negative “small granular lymphocytes” are natural killer cells. J Immunol 146: 4048–4052
Ishikawa H, Shinkai Y-I, Yagita H, Yue CC, Henkart PA, Sawada S, Young HA, Reynolds CW, Okumura K (1989): Molecular cloning of rat cytolysin. J Immunol 143: 3069–3073
Jenkison EJ, Kingston R, Smith CA, Williams GT, Owen JJT (1989): Antigen-induced apoptosis in developing T cells. A mechanism for negative selection of the T cell receptor repertoire. Eur J Immunol 19: 2175–2177
Jenne DE, Tschopp J (1988): Granzymes, a family of serine proteases released from granules of cytolytic T lymphocytes upon T cell receptor stimulation. Immunol Rev 103: 53–71
Jiang S, Ojcius DM, Persechini PM, Young JD-E (1990): Inhibition of perform binding activity by surface membrane proteins. J Immunol 144: 998–1003
Ju S-T (1991): Distinct pathways of CD4 and CD8 cells induce rapid target DNA fragmentation. J Immunol 146: 812–818
Katz P, Zaytoun AM, Fauci AS (1982): Mechanisms of human cell-mediated cytotoxicity. I. Modulation of natural killer cell activity by cyclic nucleotides. J Immunol 129: 287–296
Kranz DM, Eisen HN (1987): Resistance of cytotoxic T lymphocytes to lysin by a clone of cytotoxic T lymphocytes. Proc Natl Acad Sci USA 84: 3375–3379
Lichtenheld MG, Podack ER (1989): Structure of the human perform gene: A simple organization with interesting potential regulatory sequences. J Immunol 143: 4267–4274
Lichtenheld MG, Olsen KJ, Lu P, Lowrey DM, Hameed A, Hengartner H, Podack ER (1988): Structure and function of human perform. Nature 335: 448–451
Liu C-C, Detmers PA, Kiang S, Young JD-E (1989a): Identification and characterization of a membrane-bound cytotoxin of murine cytolytic lymphocytes that is related to tumor necrosis factor/cachectin. Proc Natl Acad Sci USA 86: 3286–3290
Liu C-C, Joag SV, Kwon BS, Young JD-E (1990): Induction of perform and serine esterase in a murine cytotoxic T lymphocyte clone. J Immunol 144: 1196–1201
Liu C-C, Rafii S, Granelli-Piperno A, Trapani JA, Young JDE (1989b): Perform and serine esterase gene expression in stimulated human T cells. J Exp Med 170: 2105–2188
Liu C-C, Steffen M, King F, Young JD-E (1987): Identification, isolation and characterization of a novel cytotoxin in murine cytolytic lymphocytes. Cell 51: 393–403
Lowrey DM, Aebischer T, Olsen K, Lichtenheld M, Rupp F, Hengartner H, Podack ER (1989): Cloning analysis and expression of murine perform 1 cDNA, a component of cytolytic T-cell granules with homology to complement component C9. Proc Natl Acad Sci USA 86: 247–251
Lu P, Garcia-Sanz JA, Lichtenheld MG, Podack ER (1990): Calcium ionophore can induce perform gene expression in human peripheral blood lymphocytes. FASEB J 4: 1898 (Abstract)
Martz E, Heagy W, Gromkowski SH (1983): The mechanism of CTL-mediated killing: Monoclonal antibody analysis of the roles of killer and target cell membrane proteins. Immunol Rev 72: 73–94
Mercep M, Weissman AM, Frank SJ, Klausner RD, Ashwell JD (1989): Activation-driven programmed cell death and T cell receptor l’rl expression. Science 246: 1162–1165
Muller C, Kagi D, Aebischer T, Odermatt B, Held W, Podack ER, Zinkernagel RM, Hengartner H (1989): Detection of perform and granzyme A mRNA in infiltrating cells during infection of mice with lymphocytic choriomeningitis virus. Eur J Immunol 19: 1253–1259
Munger WE (1988): LGL secretory granule-associated 60-kd BLT esterase augments the lytic activity of cytolysin against nucleated target cells. Nat Immun Cell Growth Regul 7: 61–62
Munger WE, Berrebi GA, Henkart PA (1988): Possible involvement of CTL granule proteases in target cell DNA breakdown. Immunol Rev 103: 99–109
Nagler-Anderson C, Allbritton NL, Verret CR, Eisen HN (1988): A comparison of the cytolytic properties of murine primary CD8’ cytotoxic T lymphocytes and cloned cytotoxic T cell lines. Immunol Rev 103: 111–124
Nagler-Anderson C, Lichtenheld M, Eisen HN, Podack ER (1989): Perform mRNA in primary peritoneal exudate cytotoxic T lymphocytes. J Immunol 143: 3440–3443
Nakata M, Smyth MJ, Norihisa Y, Kawasaki A, Shinkai Y, Okumura K, Yagita H (1990): Constitutive expression of pore-forming protein in peripheral blood 7/6 T cells: Implication for their cytotoxic role in vivo. J Exp Med 172: 1877–1880
Newell MK, Haughn LJ, Maroun CR, Julius MH (1990): Death of mature cells by separate ligation of CD4 and T-cell receptor for antigen. Nature 347: 286–289
Ojcius DM, Young JD-E (1990): Cell-mediated killing: Effector mechanisms and mediators. Cancer Cells 2: 138–145
Old LJ (1988): Tumor necrosis factor. Sci Am 258: 59–75
Ortaldo JR, Herberman RB (1984): Heterogeneity of natural killer cells. Annu Rev Immunol 2: 359–394
Ortaldo JR, Phillips W, Wasserman K, Herberman RB (1980): Effects of metabolic inhibitors on spontaneous and interferon-boosted human natural killer cell activity. J Immunol 125: 1839–1844
Ostergaard HL. Kane KP, Mescher MF, Clark WR (1987): Cytotoxic T lymphocyte mediated lysis without release of serine esterase. Nature 330: 71–72
Ottenhoff THM, Mutis T (1990): Specific killing of cytotoxic T cells and antigen-presenting cells by CD4+ cytotoxic T-cell clones. A novel potentially immunoregulatory T-T cell interaction in man. J Exp Med 171: 2011–2024
Pasternack MS, Verret CR, Liu MA, Eisen HN (1986): Serine esterase in cytolytic T lymphocytes. Nature 322: 740–743
Peters PJ, Geuze HJ, Van der Donk HA, Borst J (1990): A new model for lethal hit delivery by cytotoxic T lymphocytes. Immunol Today 11: 28–32
Peters PJ, Geuze HJ, Van der Donk HA, Slot JW, Griffith JM, Stam NJ, Clevers HC, Borst J (1989): Molecules relevant for T cell-target cell interaction are present in cytolytic granules of human T lymphocytes. Eur J Immunol 19: 1469–1475
Podack ER (1984): Molecular composition of the tubular structure of the membrane attack complex complement. J Biol Chem 259: 8641–8647
Podack ER, Dennert G (1983): Assembly of two types of tubules with putative cytolytic function by cloned natural killer cells. Nature 302: 442–445
Podack ER, Konigsberg PJ (1984): Cytolytic T cell granules: Isolation, structural, biochemical and functional characterization. J Exp Med 160: 695–710
Podack ER, Lee MK (1988): Mechanism of lymphocyte-mediated tumor cell lysis: Selective inhibition of DNA breakdown and release from target cells does not interfere with chromium release and cell death. In: Tumor Necrosis Factor/Cachectin and Related Cytokines, Bonavida B, Gifford GE, Kirchner H, Old LJ, eds. Basel: Karger
Podack ER, Lowrey DM, Lichtenheld MG, Olsen KJ, Aebischer T, Binder D, Rupp F, Hengartner H (1988): Structure, function, and expression of murine and human perforin 1 (PI). Immunol Rev 103: 203–211
Poe M, Blake JT, Boulton DA, Gammon M, Sigal NH, Wu JK, Zweerink HJ (1991): Human cytotoxic lymphocyte granzyme B. J Biol Chem 266: 98–103
Quan PC, Ishizaka T, Bloom BR (1982): Studies on the mechanism of NK cell lysis. J Immunol 128: 1786–1791
Roder JC, Haliotis TA (1980): Comparative analysis of the NK cytolytic mechanism and regulatory genes. In: Natural Cell-Mediated Immunity Against Tumors, Herberman RB, ed. New York: Academic Press
Ruddle NH, Schmid DS (1987): The role of lymphotoxin in T-cell mediated cytotoxicity. Ann Inst Pasteur Immunol 138: 314–320
Russell JH (1983): Internal disintegration model of cytotoxic lymphocyte-induced target damage. Immunol Rev 72: 97–118
Shinkai Y, Takio K, Okumura K (1988): Homology of perforin to the ninth component of complement (C9). Nature 33: 525–527
Shiver JW, Henkart PA (1991): A noncytotoxic mast cell tumor line exhibits potent IgE-dependent cytotoxicity after transfection with the cytolysin/perforin gene. Cell 64: 1175–1181
Simon MM, Hoschutzky, H, Fruth U, Simon HG, Kramer MD (1986): Purification and characterization of a T cell specific serine proteinase (TSP-1) from cloned cytolytic T lymphocytes. EMBO J 5: 3267–3274
Simon MM, Prester, M, Nerz G, Kramer MD, Fruth U (1988): Release of biologically active fragments from human plasma-fibronectin by murine T cell-specific proteinase 1 (TSP-1). Biol Chem Hoppe Seiler 369: 107–112
Smyth MJ, Norihisa Y, Gerard JR, Young HA, Ortaldo JR (1991a): IL-7 regulation of cytotoxic lymphocytes. Pore-forming protein gene expression, interferon-7 production and cytotoxicity of human peripheral blood lymphocyte subsets. Cell Immunol 138: 390–403
Smyth MJ, Norihisa Y, Ortaldo JR (1992): Multiple cytolytic mechanisms displayed by activated human peripheral blood T cell subsets. J Immunol 148: 55–62
Smyth MJ, Ortaldo JR, Bere W, Yagita H, Okumura K, Young HA (1990a): IL-2 and IL-6 synergize to augment the pore-forming protein gene expression and cytotoxic potential of human peripheral blood T cells. J Immunol 145: 1159–1166
Smyth MJ, Ortaldo JR, Shinkai Y-I, Yagita H, Nakata M, Okumura K, Young HA (1990b): Interleukin 2 induction of pore-forming protein gene expression in human peripheral blood CD8+ T cells. J Exp Med 171: 1269–1281
Smyth MJ, Strobl SL, Young HA, Ortaldo JR, Ochoa AC (1991b): Regulation of lymphokine-activated killer activity and pore-forming protein gene expression in human peripheral blood CD8+ T lymphocytes. Inhibition by transforming growth factor-fl. J Immunol 146: 3290–3297
Stevens RL, Kamada MM, Serafin WE (1988): Structure and function of the family of proteoglycans that reside in the secretory granules of natural killer cells and other effector cells of the immune response. Curr Top Microbiol Immunol 140: 93–108
Strack P, Martin C, Saito S, DeKruyff RH, Ju S-T (1990): Metabolic inhibitors distinguish cytolytic activity of CD4 and CD8 clones. Eur J Immunol 20: 179–184
Tadakuma T, Harutoshi K, Odaka, C, Kubota R, Ishimura Y, Tagita H, Okumura K (1990): CD4+, CD8+ thymocytes are susceptible to DNA fragmentation induced by phorbol ester, calcium ionophore and anti-CD3 antibody. Eur J Immunol 20: 779–784
Thompson CB, Lindsten T, Ledbetter JA, Kunkel SL, Young HA, Emerson SG, Leiden JM, June CH (1989): CD28 activation pathway regulated the production of multiple T-cell-derived lymphokines/cytokines. Proc Nall Acad Sci USA 86: 1333–1337
Trauth BC, Klas C, Peters AMJ, Matzku S, Moller P, Falk W, Debatin K-M, Krammer PH (1989): Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science 245: 301–305
Trenn G, Takayama H, Sitkovsky M (1987): Exocytosis of cytolytic granules may not be required for target cell lysis by cytotoxic T-lymphocytes. Nature 330: 72–74.
Trinchieri G (1989): The biology of natural killer cells. Adv Immunol 47: 187–376
Tschopp J (1990): Perforin-mediated target cell lysis by cytolytic T lymphocytes. Annu Rev Immunol 8: 279–302
Tschopp J, Nabholz M (1987): The role of cytoplasmic granule components in cytolytic lymphocyte-mediated cytolysis. Ann Inst Pasteur Immunol 138: 290–295
Tschopp J, Schafer S, Masson D, Peitsch MC, Heusser C (1989): Phosphorylcholine acts as a Cap-dependent receptor molecule for lymphocyte perforin. Nature 337: 272–274
Wright SC, Bonavida B (1987): Studies on the mechanism of natural killer cell-mediated cytotoxicity. VIII. Functional comparison of human natural killer cytotoxic factors with recombinant lymphotoxin and tumor necrosis factor. J Immunol 138: 1791–1798
Yamamoto RS, Ware CF, Granger GA (1986): The human LT system. XI. Identification of LT and “TNFlike” forms from stimulated natural killers, specific and non-specific cytotoxic human T cells in vitro. J Immunol 137: 1878–1884
Youn BS, Liu C-C, Kim K-K, Young JD-E, Kwon MH, Kwon BS (1991): Structure of the mouse pore-forming protein (perforin) gene: Analysis of transcription initiation site, 5’-flanking sequence, and alternative splicing of 5’ untranslated regions. J Exp Med 173: 813–822
Young JD-E, Leong LG, Liu C-C, Damiano A, Wall DA, Cohn ZA (1986): Isolation and characterization of a serine esterase from cytolytic T cell granules. Cell 47: 183–194
Young LH, Klavinskis LS, Oldstone MBA, Young JD-E (1989): In vivo expression of perforin by CD8+ lymphocytes during an acute viral infection. J Exp Med 169: 2159–2171
Zagury D, Bernard J, Thierness N, Feldman M, Berke G (1975): Isolation and characterization of individual functionally reactive cytotoxic T lymphocytes. Conjugation, killing and recycling at the single cell level. Eur J Immunol 5: 818–822
Zanovello P, Bronte V, Rosato A, Pizzo P, DiVirgilio F (1990): Responses of mouse lymphocytes to extracellular ATP. II. Extracellular ATP causes cell type-dependent lysis and DNA fragmentation. J Immunol 145: 1545–1550
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1993 Birkhäuser Boston
About this chapter
Cite this chapter
Smyth, M.J., Ortaldo, J.R. (1993). Molecular Mechanisms of Lymphocyte Cytotoxicity. In: Sitkovsky, M.V., Henkart, P.A. (eds) Cytotoxic Cells: Recognition, Effector Function, Generation, and Methods. Birkhäuser Boston. https://doi.org/10.1007/978-1-4684-6814-4_21
Download citation
DOI: https://doi.org/10.1007/978-1-4684-6814-4_21
Publisher Name: Birkhäuser Boston
Print ISBN: 978-1-4684-6816-8
Online ISBN: 978-1-4684-6814-4
eBook Packages: Springer Book Archive