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. 1980 Apr;28(1):137–146. doi: 10.1128/iai.28.1.137-146.1980

Analysis of the Lytic Step in the Herpes Simplex Virus Antibody-Dependent Cellular Cytotoxicity System

Steven L Shore 1, Thomas J Romano 1
PMCID: PMC550903  PMID: 6155344

Abstract

An antibody-dependent cellular cytotoxicity (ADCC) system in which herpes simplex virus-infected Chang liver cells are used was assessed for its dependency on cellular energy, ribonucleic acid and protein synthesis, and cytoskeletal structures such as microfilaments and microtubules. The cytotoxic reaction was only slightly inhibited when glycolysis was blocked in a glucose-free medium containing 10−2 M 2-deoxy-d-glucose. It was more substantially inhibited when respiration was blocked with 10−2 M sodium azide. The reaction was totally ablated, however, only when both glycolysis and respiration were suppressed. This inhibitory effect of energy deprivation was mediated solely at the level of the effector cell. Ribonucleic acid synthesis or protein synthesis by the effector cells was not required, as shown by the fact that neither actinomycin D, cycloheximide, nor emetine significantly inhibited ADCC. The ADCC reaction was partially inhibited by cytochalasin B, whose inhibitory effect was rapidly reversible, and was completely and irreversibly inhibited by cytochalasin A. Cytochalasin A acted on the effector cells rather than the target cells. The reaction was also partially inhibited by colchicine, whose inhibitory effect was directed solely against the effector cells and was slowly reversible. The inhibitory effects of cytochalasin B and colchicine, when used in tandem at submaximal inhibitory concentrations, were slightly more than additive. The results suggest a cooperative role for effector cell microfilaments and microtubules in mediating ADCC. Kinetic studies of ongoing herpes simplex virus ADCC reactions after initial centrifugation showed that the lytic step requires expenditure of metabolic energy as well as intact function of both microfilaments and microtubules. These findings, in concert with previous data, indicate that the ADCC process against herpes simplex virus-infected Chang liver cells can be resolved into adhesion and lytic steps. The lytic step can be readily distinguished from the adhesion step by its increased sensitivity to low ambient temperature or metabolic energy deprivation, its sensitivity to thermal inactivation, its requirements for extracellular divalent cations, and its dependence on normal function of both microfilaments and microtubules.

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Selected References

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  1. Brunner K. T., Mauel J., Cerottini J. C., Chapuis B. Quantitative assay of the lytic action of immune lymphoid cells on 51-Cr-labelled allogeneic target cells in vitro; inhibition by isoantibody and by drugs. Immunology. 1968 Feb;14(2):181–196. [PMC free article] [PubMed] [Google Scholar]
  2. Bubbers J. E., Henney C. S. Studies on the mechanism of lymphocyte-mediated cytolysis. V. The use of cytochalasins A and B to dissociate glucose transport from the lytic event. J Immunol. 1975 Jul;115(1):145–149. [PubMed] [Google Scholar]
  3. David J. R., Butterworth A. E., Remold H. G., David P. H., Houba V., Sturrock R. F. Antibody-dependent, eosinophil-mediated damage to 51Cr-labeled schistosomula of Schistosoma mansoni: effect of metabolic inhibitors and other agents which alter cell function. J Immunol. 1977 Jun;118(6):2221–2229. [PubMed] [Google Scholar]
  4. De Petris S. Inhibition and reversal of capping by cytochalasin B, vinblastine and colchicine. Nature. 1974 Jul 5;250(461):54–56. doi: 10.1038/250054a0. [DOI] [PubMed] [Google Scholar]
  5. Gately M. K., Mayer M. M., Henney C. S. Effect of anti-lymphotoxin on cell-mediated cytotoxicity. Evidence for two pathways, one involving lymphotoxin and the other requiring intimate contact between the plasma membranes of killer and target cells. Cell Immunol. 1976 Nov;27(1):82–93. doi: 10.1016/0008-8749(76)90156-8. [DOI] [PubMed] [Google Scholar]
  6. Gelfand E. W., Morris S. A., Resch K. Antibody-dependent cytotoxicity: modulation by the cytochalasins and microtubule-disruptive agents. J Immunol. 1975 Mar;114(3):919–924. [PubMed] [Google Scholar]
  7. Gillespie E., Lichtenstein L. M. Histamine release from human leukocytes: studies with deuterium oxide, colchicine, and cytochalasin B. J Clin Invest. 1972 Nov;51(11):2941–2947. doi: 10.1172/JCI107118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Henney C. S. On the mechanism of T-cell mediated cytolysis. Transplant Rev. 1973;17(0):37–70. doi: 10.1111/j.1600-065x.1973.tb00123.x. [DOI] [PubMed] [Google Scholar]
  9. Joseph B. S., Oldstone M. B. Antibody-induced redistribution of measles virus antigens on the cell surface. J Immunol. 1974 Oct;113(4):1205–1209. [PubMed] [Google Scholar]
  10. Kalina M., Berke G. Contact regions of cytotoxic T lymphocyte-target cell conjugates. Cell Immunol. 1976 Jul;25(1):41–51. doi: 10.1016/0008-8749(76)90095-2. [DOI] [PubMed] [Google Scholar]
  11. Knowles R. W., Person S. Effects of 2-deoxyglucose, glucosamine, and mannose on cell fusion and the glycoproteins of herpes simplex virus. J Virol. 1976 May;18(2):644–651. doi: 10.1128/jvi.18.2.644-651.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MacDonald H. R. Early detection of potentially lethal events in T cell-mediated cytolysis. Eur J Immunol. 1975 Apr;5(4):251–254. doi: 10.1002/eji.1830050406. [DOI] [PubMed] [Google Scholar]
  13. MacDonald H. R. Energy metabolism and T-cell-mediated cytolysis. II. Selective inhibition of cytolysis by 2-deoxy-D-glucose. J Exp Med. 1977 Sep 1;146(3):710–719. doi: 10.1084/jem.146.3.710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MacLennan I. C., Golstein P. Requirement for hexose, unrelated to energy provision, in T-cell-mediated cytolysis at the lethal hit stage. J Exp Med. 1978 Jun 1;147(6):1551–1567. doi: 10.1084/jem.147.6.1551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. MacLennan I. C., Harding B. The role of immunoglobulins in lymphocyte-mediated cell damage, in vitro. II. The mechanism of target cell damage by lymphoid cells from immunized rats. Immunology. 1970 Mar;18(3):405–412. [PMC free article] [PubMed] [Google Scholar]
  16. Martz E., Benacerraf B. An effector-cell independent step in target cell lysis by sensitized mouse lymphocytes. J Immunol. 1973 Nov;111(5):1538–1545. [PubMed] [Google Scholar]
  17. Michl J., Ohlbaum D. J., Silverstein S. C. 2-Deoxyglucose selectively inhibits Fc and complement receptor-mediated phagocytosis in mouse peritoneal macrophages. I. Description of the inhibitory effect. J Exp Med. 1976 Dec 1;144(6):1465–1483. doi: 10.1084/jem.144.6.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nicolson G. L., Poste G. The cancer cell: dynamic aspects and modifications in cell-surface organization (first of two parts). N Engl J Med. 1976 Jul 22;295(4):197–203. doi: 10.1056/NEJM197607222950405. [DOI] [PubMed] [Google Scholar]
  19. Romano T. J., Shore S. L. Analysis of the adhesion step in the herpes simplex virus antibody-dependent cellular cytotoxicity system. Infect Immun. 1979 Oct;26(1):163–172. doi: 10.1128/iai.26.1.163-172.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Romano T. J., Shore S. L. Lysis of virus-infected target cells by antibody-dependent cellular cytotoxicity. I. General requirements of the reaction and temporal relationship between lethal hits and cytolysis. Cell Immunol. 1977 Apr;30(1):66–81. doi: 10.1016/0008-8749(77)90048-x. [DOI] [PubMed] [Google Scholar]
  21. Shore S. L., Black C. M., Melewicz F. M., Wood P. A., Nahmias A. J. Antibody-dependent cell-mediated cytotoxicity to target cells infected with type 1 and type 2 herpes simplex virus. J Immunol. 1976 Jan;116(1):194–201. [PubMed] [Google Scholar]
  22. Shore S. L., Melewicz F. M., Gordon D. S. The mononuclear cell in human blood which mediates antibody-dependent cellular cytotoxicity to virus-infected target cells. I. Identification of the population of effector cells. J Immunol. 1977 Feb;118(2):558–566. [PubMed] [Google Scholar]
  23. Shore S. L., Melewicz F. M., Milgrom H., Nahmias A. J. Antibody-dependent cell-mediated cytotoxicity to target cells infected with herpes simplex viruses. Adv Exp Med Biol. 1976;73(Pt B):217–227. doi: 10.1007/978-1-4684-3300-5_19. [DOI] [PubMed] [Google Scholar]
  24. Shore S. L., Nahmias A. J., Starr S. E., Wood P. A., McFarlin D. E. Detection of cell-dependent cytotoxic antibody to cells infected with herpes simplex virus. Nature. 1974 Sep 27;251(5473):350–352. doi: 10.1038/251350a0. [DOI] [PubMed] [Google Scholar]
  25. Shore S. L., Romano T. J., Hubbard M. R., Gordon D. S. Lysis of virus-infected target cells by antibody-dependent cellular cytotoxicity. II. Reversibility of the heart inactivation of K cell killing and relationship of Fc receptor capping to lysis. Cell Immunol. 1977 Jun 1;31(1):55–61. doi: 10.1016/0008-8749(77)90006-5. [DOI] [PubMed] [Google Scholar]
  26. Strom T. B., Garovoy M. R., Bear R. A., Gribik M., Carpenter C. B. A comparison of the effects of metabolic inhibitors upon direct and antibody-dependent lymphocyte mediated cytotoxicity. Cell Immunol. 1975 Dec;20(2):247–256. doi: 10.1016/0008-8749(75)90102-1. [DOI] [PubMed] [Google Scholar]
  27. Strom T. B., Garovoy M. R., Carpenter D. B., Merrill J. P. Microtubule function in immune and nonimmune lymphocyte-mediated cytotoxicity. Science. 1973 Jul 13;181(4095):171–173. doi: 10.1126/science.181.4095.171. [DOI] [PubMed] [Google Scholar]
  28. Thorn R. M., Henney C. S. Studies on the mechanism of lymphocyte-mediated cytolysis. VI. A reappraisal of the requirement for protein synthesis during T cell-mediated lysis. J Immunol. 1976 Jan;116(1):146–149. [PubMed] [Google Scholar]
  29. Trinchieri G., Bauman P., De Marchi M., Tökés Z. Antibody-dependent cell-mediated cytotoxicity in humans. I. Characterization of the effector cell. J Immunol. 1975 Jul;115(1):249–255. [PubMed] [Google Scholar]
  30. Trinchieri G., De Marchi M. Antibody-dependent cell-mediated cytotoxicity in humans. II. Energy requirement. J Immunol. 1975 Jul;115(1):256–260. [PubMed] [Google Scholar]
  31. Trinchieri G., De Marchi M. Antibody-dependent cell-mediated cytotoxicity in humans. III. Effect of protease inhibitors and substrates. J Immunol. 1976 Apr;116(4):885–891. [PubMed] [Google Scholar]
  32. Wessells N. K., Spooner B. S., Ash J. F., Bradley M. O., Luduena M. A., Taylor E. L., Wrenn J. T., Yamada K. Microfilaments in cellular and developmental processes. Science. 1971 Jan 15;171(3967):135–143. doi: 10.1126/science.171.3967.135. [DOI] [PubMed] [Google Scholar]
  33. Ziegler H. K., Geyer C., Henney C. S. Studies on the cytolytic activity of human lymphocytes. III. An analysis of requirements for K cell-target interaction. J Immunol. 1977 Nov;119(5):1821–1829. [PubMed] [Google Scholar]

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