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J Virol. 1990 August; 64(8): 3726–3733.
PMCID: PMC249667

Mechanism of killing by virus-induced cytotoxic T lymphocytes elicited in vivo.

Abstract

The mechanism of lysis by in vivo-induced cytotoxic T lymphocytes (CTL) was examined with virus-specific CTL from mice infected with lymphocytic choriomeningitis virus (LCMV). LCMV-induced T cells were shown to have greater than 10 times the serine esterase activity of T cells from normal mice, and high levels of serine esterase were located in the LCMV-induced CD8+ cell population. Serine esterase was also induced in purified T-cell preparations isolated from mice infected with other viruses (mouse hepatitis, Pichinde, and vaccinia). In contrast, the interferon inducer poly(I.C) only marginally enhanced serine esterase in T cells. Serine esterase activity was released from the LCMV-induced T cells upon incubation with syngeneic but not allogeneic LCMV-infected target cells. Both cytotoxicity and the release of serine esterase were calcium dependent. Serine esterase released from disrupted LCMV-induced T cells was in the form of the fast-sedimenting particles, suggesting its inclusion in granules. Competitive substrates for serine esterase blocked killing by LCMV-specific CTL, but serine esterase-containing granules isolated from LCMV-induced CTL, in contrast to granules isolated from a rat natural killer cell tumor line, did not display detectable hemolytic activity. Fragmentation of target cell DNA was observed during the lytic process mediated by LCMV-specific CTL, and the release of the DNA label [125I]iododeoxyuridine from target cells and the accompanying fragmentation of DNA also were calcium dependent. These data support the hypothesis that the mechanism of killing by in vivo-induced T cells involves a calcium-dependent secretion of serine esterase-containing granules and a target cell death by a process involving nuclear degradation and DNA fragmentation.

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Selected References

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  • Berke G, Rosen D. Highly lytic in vivo primed cytolytic T lymphocytes devoid of lytic granules and BLT-esterase activity acquire these constituents in the presence of T cell growth factors upon blast transformation in vitro. J Immunol. 1988 Sep 1;141(5):1429–1436. [PubMed]
  • Biron CA, Habu S, Okumura K, Welsh RM. Lysis of uninfected and virus-infected cells in vivo: a rejection mechanism in addition to that mediated by natural killer cells. J Virol. 1984 Jun;50(3):698–707. [PMC free article] [PubMed]
  • Biron CA, Natuk RJ, Welsh RM. Generation of large granular T lymphocytes in vivo during viral infection. J Immunol. 1986 Mar 15;136(6):2280–2286. [PubMed]
  • Biron CA, Pedersen KF, Welsh RM. Aberrant T cells in beige mutant mice. J Immunol. 1987 Apr 1;138(7):2050–2056. [PubMed]
  • Dennert G, Anderson CG, Prochazka G. High activity of N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester serine esterase and cytolytic perforin in cloned cell lines is not demonstrable in in-vivo-induced cytotoxic effector cells. Proc Natl Acad Sci U S A. 1987 Jul;84(14):5004–5008. [PubMed]
  • Duke RC, Chervenak R, Cohen JJ. Endogenous endonuclease-induced DNA fragmentation: an early event in cell-mediated cytolysis. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6361–6365. [PubMed]
  • Habu S, Fukui H, Shimamura K, Kasai M, Nagai Y, Okumura K, Tamaoki N. In vivo effects of anti-asialo GM1. I. Reduction of NK activity and enhancement of transplanted tumor growth in nude mice. J Immunol. 1981 Jul;127(1):34–38. [PubMed]
  • Hameed A, Olsen KJ, Lee MK, Lichtenheld MG, Podack ER. Cytolysis by Ca-permeable transmembrane channels. Pore formation causes extensive DNA degradation and cell lysis. J Exp Med. 1989 Mar 1;169(3):765–777. [PMC free article] [PubMed]
  • Jones CL, Simpson NJ, Waryas VL, Pappas MG. An inexpensive method of drying multiple agarose gels. Biotechniques. 1988 Feb;6(2):118–122. [PubMed]
  • Julius MH, Simpson E, Herzenberg LA. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973 Oct;3(10):645–649. [PubMed]
  • Kramer MD, Fruth U, Simon HG, Simon MM. Expression of cytoplasmic granules with T cell-associated serine proteinase-1 activity in Ly-2+(CD8+) T lymphocytes responding to lymphocytic choriomeningitis virus in vivo. Eur J Immunol. 1989 Jan;19(1):151–156. [PubMed]
  • Lichtenheld MG, Olsen KJ, Lu P, Lowrey DM, Hameed A, Hengartner H, Podack ER. Structure and function of human perforin. Nature. 1988 Sep 29;335(6189):448–451. [PubMed]
  • Lowrey DM, Aebischer T, Olsen K, Lichtenheld M, Rupp F, Hengartner H, Podack ER. Cloning, analysis, and expression of murine perforin 1 cDNA, a component of cytolytic T-cell granules with homology to complement component C9. Proc Natl Acad Sci U S A. 1989 Jan;86(1):247–251. [PubMed]
  • Lutzner MA, Lowrie CT, Jordan HW. Giant granules in leukocytes of the beige mouse. J Hered. 1967 Nov-Dec;58(6):299–300. [PubMed]
  • Martz E, Howell DM. CTL: virus control cells first and cytolytic cells second? DNA fragmentation, apoptosis and the prelytic halt hypothesis. Immunol Today. 1989 Mar;10(3):79–86. [PubMed]
  • Martz E, Parker WL, Gately MK, Tsoukas CD. The role of calcium in the lethal hit of T lymphocyte-mediated cytolysis. Adv Exp Med Biol. 1982;146:121–147. [PubMed]
  • Masson D, Tschopp J. Isolation of a lytic, pore-forming protein (perforin) from cytolytic T-lymphocytes. J Biol Chem. 1985 Aug 5;260(16):9069–9072. [PubMed]
  • McIntyre KW, Natuk RJ, Biron CA, Kase K, Greenberger J, Welsh RM. Blastogenesis of large granular lymphocytes in nonlymphoid organs. J Leukoc Biol. 1988 Jun;43(6):492–501. [PubMed]
  • McIntyre KW, Welsh RM. Accumulation of natural killer and cytotoxic T large granular lymphocytes in the liver during virus infection. J Exp Med. 1986 Nov 1;164(5):1667–1681. [PMC free article] [PubMed]
  • Millard PJ, Henkart MP, Reynolds CW, Henkart PA. Purification and properties of cytoplasmic granules from cytotoxic rat LGL tumors. J Immunol. 1984 Jun;132(6):3197–3204. [PubMed]
  • Munger WE, Berrebi GA, Henkart PA. Possible involvement of CTL granule proteases in target cell DNA breakdown. Immunol Rev. 1988 Mar;103:99–109. [PubMed]
  • Ostergaard HL, Kane KP, Mescher MF, Clark WR. Cytotoxic T lymphocyte mediated lysis without release of serine esterase. Nature. 1987 Nov 5;330(6143):71–72. [PubMed]
  • Pasternack MS, Eisen HN. A novel serine esterase expressed by cytotoxic T lymphocytes. Nature. 314(6013):743–745. [PubMed]
  • Russell JH. Internal disintegration model of cytotoxic lymphocyte-induced target damage. Immunol Rev. 1983;72:97–118. [PubMed]
  • Scuderi P. Suppression of human leukocyte tumor necrosis factor secretion by the serine protease inhibitor p-toluenesulfonyl-L-arginine methyl ester (TAME). J Immunol. 1989 Jul 1;143(1):168–173. [PubMed]
  • Simon MM, Fruth U, Simon HG, Kramer MD. A specific serine proteinase is inducible in Lyt-2+,L3T4- and Lyt-2-,L3T4+ T cells in vitro but is mainly associated with Lyt-2+,L3T4- effector cells in vivo. Eur J Immunol. 1986 Dec;16(12):1559–1568. [PubMed]
  • Townsend AR, Rothbard J, Gotch FM, Bahadur G, Wraith D, McMichael AJ. The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell. 1986 Mar 28;44(6):959–968. [PubMed]
  • Trenn G, Takayama H, Sitkovsky MV. Exocytosis of cytolytic granules may not be required for target cell lysis by cytotoxic T-lymphocytes. Nature. 1987 Nov 5;330(6143):72–74. [PubMed]
  • Utsunomiya N, Nakanishi M. A serine protease triggers the initial step of transmembrane signalling in cytotoxic T cells. J Biol Chem. 1986 Dec 15;261(35):16514–16517. [PubMed]
  • Welsh RM., Jr Cytotoxic cells induced during lymphocytic choriomeningitis virus infection of mice. I. Characterization of natural killer cell induction. J Exp Med. 1978 Jul 1;148(1):163–181. [PMC free article] [PubMed]
  • Welsh RM. Regulation and role of large granular lymphocytes in arenavirus infections. Curr Top Microbiol Immunol. 1987;134:185–209. [PubMed]
  • Yang HY, Dundon PL, Nahill SR, Welsh RM. Virus-induced polyclonal cytotoxic T lymphocyte stimulation. J Immunol. 1989 Mar 1;142(5):1710–1718. [PubMed]
  • Yang H, Welsh RM. Induction of alloreactive cytotoxic T cells by acute virus infection of mice. J Immunol. 1986 Feb 15;136(4):1186–1193. [PubMed]
  • Young JD, Liu CC. Multiple mechanisms of lymphocyte-mediated killing. Immunol Today. 1988 May;9(5):140–144. [PubMed]
  • Young LH, Klavinskis LS, Oldstone MB, Young JD. In vivo expression of perforin by CD8+ lymphocytes during an acute viral infection. J Exp Med. 1989 Jun 1;169(6):2159–2171. [PMC free article] [PubMed]
  • Zinkernagel RM, Doherty PC. MHC-restricted cytotoxic T cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function, and responsiveness. Adv Immunol. 1979;27:51–177. [PubMed]

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