Analysis of Sequential Substages of the Natural Killer Cell Lethal Hit

  • Richard L. Deem
  • Stephan R. Targan
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 187)


Previous studies have shown that NK cytotoxicity can be resolved into several stages: NK-target cell binding, triggering, programming, and killer cell independent lysis (KCIL)(1, 2). NK cells produce soluble cytolytic factors (NKCF), which can lyse NK-sensitive targets and have been used as another measure of the NK lethal hit (3, 4). Since KCIL and NKCF-mediated cytolysis are independent of the NK cell, and are inhibited by PGE, (5), low temperatue (1), and trypsin (6), it was postulated that the target cell membrane probably played a crucial role during the NK lethal hit. Several hypotheses were proposed to explain these results. [1] Membrane fluidity/movement may play a role in channel formation by the NK lytic complex. [2] Membrane movement or endocytosis, requiring intact metabolic and energy transfer pathways, may be required for completion of the lethal hit. [3] Enzymatic activity, either from target cell enzymes or a function of the NK lytic complex itself, may be a necessary substage of the NK lethal hit. These hypotheses were tested in this study with specific membrane crosslinking/fluidizing agents, inhibitors of energy metabolism, and inhibitors of protease activity.


Natural Killer Benzyl Alcohol Crosslinking Reagent Serine Protease Activity Target Cell Membrane 
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  1. 1.
    Hiserodt, J.C., L.J. Britvan, and S.R. Targan. 1982. Characterization of the cytolytic reaction mechanism of the human natural killer (NK) lymphocyte: resolution into binding, programming, and killer cell-independent steps. J. Immunol. 129: 1782.PubMedGoogle Scholar
  2. 2.
    Targan, S.R. and W. Newman. 1983. Definition of a “Trigger” stage in the NK cytolytic reaction sequence by a monoclonal antibody to the glycoprotein T-200. J. Immunol. 131: 1149.PubMedGoogle Scholar
  3. 3.
    Wright, S.C. and B. Bonavida. 1982. Studies on the mechanism of natural killer (NK) cell-mediated cytotoxicity (CMC). I. Release of cytotoxic factors specific for NK-sensitive target cells (NKCF) during co-culture of NK effector cells with NK target cells. J. Immunol. 129: 433.PubMedGoogle Scholar
  4. 4.
    Farrum, E. and S.R. Targan. 1983. Identification of human natural killer soluble factors (NKCF) derived from NK-enriched lymphocyte populations: specificity of generation and killing. J. Immunol. 130: 1252.Google Scholar
  5. 5.
    Hiserodt, J.C., L. Britvan, and S.R. Targan. 1982. Differential effects of various pharmocologic agents on the cytolytic reaction mechanism of the human natural killer lymphocyte: further resolution of programming for lysis and KCIL into discrete stages. J. Immunol. 129: 2266.PubMedGoogle Scholar
  6. 6.
    Hiserodt, J.C., L. Britvan, and S.R. Targan. 1983. Studies on the mechanism of the human natural killer cell lethal hit: analysis of the mechanism of protease inhibition of the lethal hit. J. Immunol. 131: 2705.PubMedGoogle Scholar
  7. 7.
    Deem, R.L. and S.R. Targan. 1984. Sequential substages of natural killer cell-derived cytolytic factor (NKCF)-mediated cytolysis as defined by glutaraldehyde modulation of the target cell. J. Immunol. 133: in press.Google Scholar
  8. 8.
    Deem, R.L. and S.R. Targan. 1984, Evidence of a dynamic role of the target cell membrane during the early stages of the natural killer cell lethal hit. J. Immunol. 133: in press.Google Scholar
  9. 9.
    Devlin, J.J., R.S. Yamamoto, and G.A. Granger. 1981. Stabilization and functional studies of high-molecular weight murine lymphotoxins. Cell. Immunol. 61: 22.PubMedCrossRefGoogle Scholar
  10. 10.
    Bowes, J.H. and C.W. Cater. 1965. The reaction of glutaralde- hyde with proteins and other biological materials. J. Roy. Micro. Soc. 85: 193CrossRefGoogle Scholar
  11. 11.
    Flitney, F.W. 1965. The time course of fixation of albumin by formaldehyde, glutaraldehyde, acrolein and other higher aldehydes. J. Roy. Micro. Soc. 85:353.CrossRefGoogle Scholar
  12. 12.
    Richards, F.M. and J.R. Knowles. 1968. Glutaraldehyde as a protein cross-linking reagent. J. Mol. Biol. 37: 231.PubMedCrossRefGoogle Scholar
  13. 13.
    Peters, K. and F.M. Knowles. 1977. Chemical cross-linking: reagents and problems in studies of membrane structure. Ann. Rev. Biochem. 46: 523.PubMedCrossRefGoogle Scholar
  14. 14.
    Hopwood, D. 1969. Fixation of proteins by osmium tetroxide, potassium dichromate and potassium permanganate. Histochimie 18: 250PubMedCrossRefGoogle Scholar
  15. 15.
    Bahr, G.F. 1954. Osmium tetroxide and ruthenium tetroxide and their reactions with biologically important substances. Exptl. Cell Res. 7: 457.PubMedCrossRefGoogle Scholar
  16. 16.
    Goldstein, I.J. and C.E. Hayes. 1976. The lectins: carbohydrate-binding proteins of plants and animals. Arch. Biochem. Biophys. 173: 127.Google Scholar
  17. 17.
    Silverstein, S.C., R.M. Steinman, and Z.A. Cohn. 1977. Endocytosis. Ann. Rev. Biochem. 46: 669.PubMedCrossRefGoogle Scholar
  18. 18.
    Chang, T.W. and H.E. Eisen. 1980. Effects of N-tosyl-L-lysyl- chloromethylketone on the activity of cytotoxic T lymphocytes. J. Immunol. 124: 1028.PubMedGoogle Scholar
  19. 19.
    Trinchieri, G. and M. DeMarchi. 1976. Antibody dependent cell-mediated cytotoxicity in humans. III. Effect of protease inhibitors and substrates. J. Immunol. 116: 885.PubMedGoogle Scholar
  20. 20.
    T. Shibata, Y. Sugiura, and S. Iwayanagi. 1982. Effects of benzyl alcohol on phosphatidylcholine lamellar phase with different water contents. Chem. Phys. Lipids 31: 105.PubMedCrossRefGoogle Scholar
  21. 21.
    Podack, E.R. and G. Dennert. 1983. Assembly of two types of tubules with putative cytolytic function by cloned natural killer cells. Nature 302: 442.PubMedCrossRefGoogle Scholar
  22. 22.
    Needham, L., A.D. Whetton, and M.D. Houslay. 1982. The local anaesthetic and bilayer fluidizing agent, benzyl alcohol decreases the thermostability of the integral membrane protein adenylate cyclase. FEBS Lett. 140: 85.PubMedCrossRefGoogle Scholar
  23. 23.
    Tolleshaug, H. and T. Berg. 1982. Evidence for the selective inhibition of fusion between endocytic vesicles and lysosomes by benzyl alcohol. Biochem. Pharmac. 31: 593.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Richard L. Deem
    • 1
    • 2
    • 3
    • 4
  • Stephan R. Targan
    • 1
    • 2
    • 3
    • 4
  1. 1.Geriatric Research Education and Clinical CenterWadsworth Veterans Administration Medical CenterLos AngelesUSA
  2. 2.Department of Medicine University of CaliforniaLos AngelesUSA
  3. 3.Department of MicrobiologyUniversity of CaliforniaLos AngelesUSA
  4. 4.Department of ImmunologyUniversity of CaliforniaLos AngelesUSA

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