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. 1997 Apr;65(4):1486–1496. doi: 10.1128/iai.65.4.1486-1496.1997

Human monocyte-derived macrophages infected with virulent Shigella flexneri in vitro undergo a rapid cytolytic event similar to oncosis but not apoptosis.

C M Fernandez-Prada 1, D L Hoover 1, B D Tall 1, M M Venkatesan 1
PMCID: PMC175157  PMID: 9119491

Abstract

Infection of human monocyte-derived macrophages in vitro with virulent Shigella flexneri resulted in cell death which involved rupture of the plasma membrane, cell swelling, disintegration of ultrastructure, and generalized karyolysis. These features bore resemblance to oncosis and are in striking contrast to previously described observations of mouse macrophages, where a similar infection by virulent Shigella resulted in cell death by apoptosis. Cell death by oncosis in human macrophages was confirmed by lactate dehydrogenase release, light microscopy, electron microscopy, terminal deoxynucleotidyltransferase end labeling of DNA ends, DNA fragmentation assays, and fluorescence-activated cell sorter analysis of propidium-labeled nuclei. Thus, the phenomena of cell death induced by virulent Shigella in human and mouse macrophages reflect different biochemical pathways. Interleukin-1beta (IL-1beta) was released in culture supernatants of human macrophages infected with virulent bacteria. Inhibition with IL-1beta-converting enzyme inhibitors indicated, however, that this release occurred as a passive event of cell lysis. The patterns of intracellular survival of Shigella strains within human and mouse macrophages reflect differences that exist not only between Shigella serotypes but also between the two different macrophage cell types.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alpuche-Aranda C. M., Racoosin E. L., Swanson J. A., Miller S. I. Salmonella stimulate macrophage macropinocytosis and persist within spacious phagosomes. J Exp Med. 1994 Feb 1;179(2):601–608. doi: 10.1084/jem.179.2.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bermudez L. E. Differential mechanisms of intracellular killing of Mycobacterium avium and Listeria monocytogenes by activated human and murine macrophages. The role of nitric oxide. Clin Exp Immunol. 1993 Feb;91(2):277–281. doi: 10.1111/j.1365-2249.1993.tb05895.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bär P. R. Apoptosis--the cell's silent exit. Life Sci. 1996;59(5-6):369–378. doi: 10.1016/0024-3205(96)00315-3. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Faherty D. A., Claudy V., Plocinski J. M., Kaffka K., Kilian P., Thompson R. C., Benjamin W. R. Failure of IL-1 receptor antagonist and monoclonal anti-IL-1 receptor antibody to inhibit antigen-specific immune responses in vivo. J Immunol. 1992 Feb 1;148(3):766–771. [PubMed] [Google Scholar]
  6. Fearnhead H. O., Dinsdale D., Cohen G. M. An interleukin-1 beta-converting enzyme-like protease is a common mediator of apoptosis in thymocytes. FEBS Lett. 1995 Nov 20;375(3):283–288. doi: 10.1016/0014-5793(95)01228-7. [DOI] [PubMed] [Google Scholar]
  7. Gordon S., Lawson L., Rabinowitz S., Crocker P. R., Morris L., Perry V. H. Antigen markers of macrophage differentiation in murine tissues. Curr Top Microbiol Immunol. 1992;181:1–37. doi: 10.1007/978-3-642-77377-8_1. [DOI] [PubMed] [Google Scholar]
  8. Granowitz E. V., Porat R., Mier J. W., Pribble J. P., Stiles D. M., Bloedow D. C., Catalano M. A., Wolff S. M., Dinarello C. A. Pharmacokinetics, safety and immunomodulatory effects of human recombinant interleukin-1 receptor antagonist in healthy humans. Cytokine. 1992 Sep;4(5):353–360. doi: 10.1016/1043-4666(92)90078-6. [DOI] [PubMed] [Google Scholar]
  9. Green S. J., Nacy C. A., Meltzer M. S. Cytokine-induced synthesis of nitrogen oxides in macrophages: a protective host response to Leishmania and other intracellular pathogens. J Leukoc Biol. 1991 Jul;50(1):93–103. doi: 10.1002/jlb.50.1.93. [DOI] [PubMed] [Google Scholar]
  10. Hale T. L. Genetic basis of virulence in Shigella species. Microbiol Rev. 1991 Jun;55(2):206–224. doi: 10.1128/mr.55.2.206-224.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henkart P. A. ICE family proteases: mediators of all apoptotic cell death? Immunity. 1996 Mar;4(3):195–201. doi: 10.1016/s1074-7613(00)80428-8. [DOI] [PubMed] [Google Scholar]
  12. High N., Mounier J., Prévost M. C., Sansonetti P. J. IpaB of Shigella flexneri causes entry into epithelial cells and escape from the phagocytic vacuole. EMBO J. 1992 May;11(5):1991–1999. doi: 10.1002/j.1460-2075.1992.tb05253.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hockenbery D. Defining apoptosis. Am J Pathol. 1995 Jan;146(1):16–19. [PMC free article] [PubMed] [Google Scholar]
  14. Jung H. C., Eckmann L., Yang S. K., Panja A., Fierer J., Morzycka-Wroblewska E., Kagnoff M. F. A distinct array of proinflammatory cytokines is expressed in human colon epithelial cells in response to bacterial invasion. J Clin Invest. 1995 Jan;95(1):55–65. doi: 10.1172/JCI117676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kelsall B. L., Strober W. The role of dendritic cells in antigen processing in the Peyer's patch. Ann N Y Acad Sci. 1996 Feb 13;778:47–54. doi: 10.1111/j.1749-6632.1996.tb21113.x. [DOI] [PubMed] [Google Scholar]
  16. Keren D. F., McDonald R. A., Wassef J. S., Armstrong L. R., Brown J. E. The enteric immune response to shigella antigens. Curr Top Microbiol Immunol. 1989;146:213–223. doi: 10.1007/978-3-642-74529-4_23. [DOI] [PubMed] [Google Scholar]
  17. Keren D. F. Mucosal IgA elaboration. Crit Rev Clin Lab Sci. 1989;27(2):159–177. doi: 10.3109/10408368909106592. [DOI] [PubMed] [Google Scholar]
  18. Labrec E. H., Schneider H., Magnani T. J., Formal S. B. EPITHELIAL CELL PENETRATION AS AN ESSENTIAL STEP IN THE PATHOGENESIS OF BACILLARY DYSENTERY. J Bacteriol. 1964 Nov;88(5):1503–1518. doi: 10.1128/jb.88.5.1503-1518.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lazebnik Y. A., Kaufmann S. H., Desnoyers S., Poirier G. G., Earnshaw W. C. Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature. 1994 Sep 22;371(6495):346–347. doi: 10.1038/371346a0. [DOI] [PubMed] [Google Scholar]
  20. Majno G., Joris I. Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol. 1995 Jan;146(1):3–15. [PMC free article] [PubMed] [Google Scholar]
  21. Mangan D. F., Mergenhagen S. E., Wahl S. M. Apoptosis in human monocytes: possible role in chronic inflammatory diseases. J Periodontol. 1993 May;64(5 Suppl):461–466. [PubMed] [Google Scholar]
  22. Mangan D. F., Robertson B., Wahl S. M. IL-4 enhances programmed cell death (apoptosis) in stimulated human monocytes. J Immunol. 1992 Mar 15;148(6):1812–1816. [PubMed] [Google Scholar]
  23. Mangan D. F., Wahl S. M. Differential regulation of human monocyte programmed cell death (apoptosis) by chemotactic factors and pro-inflammatory cytokines. J Immunol. 1991 Nov 15;147(10):3408–3412. [PubMed] [Google Scholar]
  24. Mangan D. F., Welch G. R., Wahl S. M. Lipopolysaccharide, tumor necrosis factor-alpha, and IL-1 beta prevent programmed cell death (apoptosis) in human peripheral blood monocytes. J Immunol. 1991 Mar 1;146(5):1541–1546. [PubMed] [Google Scholar]
  25. McIntyre K. W., Stepan G. J., Kolinsky K. D., Benjamin W. R., Plocinski J. M., Kaffka K. L., Campen C. A., Chizzonite R. A., Kilian P. L. Inhibition of interleukin 1 (IL-1) binding and bioactivity in vitro and modulation of acute inflammation in vivo by IL-1 receptor antagonist and anti-IL-1 receptor monoclonal antibody. J Exp Med. 1991 Apr 1;173(4):931–939. doi: 10.1084/jem.173.4.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mounier J., Vasselon T., Hellio R., Lesourd M., Sansonetti P. J. Shigella flexneri enters human colonic Caco-2 epithelial cells through the basolateral pole. Infect Immun. 1992 Jan;60(1):237–248. doi: 10.1128/iai.60.1.237-248.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Munn D. H., Beall A. C., Song D., Wrenn R. W., Throckmorton D. C. Activation-induced apoptosis in human macrophages: developmental regulation of a novel cell death pathway by macrophage colony-stimulating factor and interferon gamma. J Exp Med. 1995 Jan 1;181(1):127–136. doi: 10.1084/jem.181.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nicoletti I., Migliorati G., Pagliacci M. C., Grignani F., Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271–279. doi: 10.1016/0022-1759(91)90198-o. [DOI] [PubMed] [Google Scholar]
  29. Noriega F. R., Liao F. M., Formal S. B., Fasano A., Levine M. M. Prevalence of Shigella enterotoxin 1 among Shigella clinical isolates of diverse serotypes. J Infect Dis. 1995 Nov;172(5):1408–1410. doi: 10.1093/infdis/172.5.1408. [DOI] [PubMed] [Google Scholar]
  30. Oberhammer F., Wilson J. W., Dive C., Morris I. D., Hickman J. A., Wakeling A. E., Walker P. R., Sikorska M. Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J. 1993 Sep;12(9):3679–3684. doi: 10.1002/j.1460-2075.1993.tb06042.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Parsot C. Shigella flexneri: genetics of entry and intercellular dissemination in epithelial cells. Curr Top Microbiol Immunol. 1994;192:217–241. doi: 10.1007/978-3-642-78624-2_10. [DOI] [PubMed] [Google Scholar]
  32. Perdomo O. J., Cavaillon J. M., Huerre M., Ohayon H., Gounon P., Sansonetti P. J. Acute inflammation causes epithelial invasion and mucosal destruction in experimental shigellosis. J Exp Med. 1994 Oct 1;180(4):1307–1319. doi: 10.1084/jem.180.4.1307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rappolee D. A., Werb Z. Macrophage-derived growth factors. Curr Top Microbiol Immunol. 1992;181:87–140. doi: 10.1007/978-3-642-77377-8_4. [DOI] [PubMed] [Google Scholar]
  35. Raqib R., Lindberg A. A., Wretlind B., Bardhan P. K., Andersson U., Andersson J. Persistence of local cytokine production in shigellosis in acute and convalescent stages. Infect Immun. 1995 Jan;63(1):289–296. doi: 10.1128/iai.63.1.289-296.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Raqib R., Wretlind B., Andersson J., Lindberg A. A. Cytokine secretion in acute shigellosis is correlated to disease activity and directed more to stool than to plasma. J Infect Dis. 1995 Feb;171(2):376–384. doi: 10.1093/infdis/171.2.376. [DOI] [PubMed] [Google Scholar]
  37. Roth P., Stanley E. R. The biology of CSF-1 and its receptor. Curr Top Microbiol Immunol. 1992;181:141–167. doi: 10.1007/978-3-642-77377-8_5. [DOI] [PubMed] [Google Scholar]
  38. Sansonetti P. J., Arondel J., Cantey J. R., Prévost M. C., Huerre M. Infection of rabbit Peyer's patches by Shigella flexneri: effect of adhesive or invasive bacterial phenotypes on follicle-associated epithelium. Infect Immun. 1996 Jul;64(7):2752–2764. doi: 10.1128/iai.64.7.2752-2764.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sansonetti P. J., Arondel J., Cavaillon J. M., Huerre M. Role of interleukin-1 in the pathogenesis of experimental shigellosis. J Clin Invest. 1995 Aug;96(2):884–892. doi: 10.1172/JCI118135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Schindler R., Clark B. D., Dinarello C. A. Dissociation between interleukin-1 beta mRNA and protein synthesis in human peripheral blood mononuclear cells. J Biol Chem. 1990 Jun 25;265(18):10232–10237. [PubMed] [Google Scholar]
  41. Tomei L. D., Shapiro J. P., Cope F. O. Apoptosis in C3H/10T1/2 mouse embryonic cells: evidence for internucleosomal DNA modification in the absence of double-strand cleavage. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):853–857. doi: 10.1073/pnas.90.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wassef J. S., Keren D. F., Mailloux J. L. Role of M cells in initial antigen uptake and in ulcer formation in the rabbit intestinal loop model of shigellosis. Infect Immun. 1989 Mar;57(3):858–863. doi: 10.1128/iai.57.3.858-863.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wyllie A. H. Apoptosis: cell death in tissue regulation. J Pathol. 1987 Dec;153(4):313–316. doi: 10.1002/path.1711530404. [DOI] [PubMed] [Google Scholar]
  44. Yamamoto Y., Klein T. W., Friedman H. Legionella and macrophages. Immunol Ser. 1994;60:329–348. [PubMed] [Google Scholar]
  45. Zakeri Z. F., Quaglino D., Latham T., Lockshin R. A. Delayed internucleosomal DNA fragmentation in programmed cell death. FASEB J. 1993 Mar;7(5):470–478. doi: 10.1096/fasebj.7.5.8462789. [DOI] [PubMed] [Google Scholar]
  46. Zhu H., Fearnhead H. O., Cohen G. M. An ICE-like protease is a common mediator of apoptosis induced by diverse stimuli in human monocytic THP.1 cells. FEBS Lett. 1995 Oct 30;374(2):303–308. doi: 10.1016/0014-5793(95)01116-v. [DOI] [PubMed] [Google Scholar]
  47. Zychlinsky A., Fitting C., Cavaillon J. M., Sansonetti P. J. Interleukin 1 is released by murine macrophages during apoptosis induced by Shigella flexneri. J Clin Invest. 1994 Sep;94(3):1328–1332. doi: 10.1172/JCI117452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Zychlinsky A., Kenny B., Ménard R., Prévost M. C., Holland I. B., Sansonetti P. J. IpaB mediates macrophage apoptosis induced by Shigella flexneri. Mol Microbiol. 1994 Feb;11(4):619–627. doi: 10.1111/j.1365-2958.1994.tb00341.x. [DOI] [PubMed] [Google Scholar]
  49. Zychlinsky A., Prevost M. C., Sansonetti P. J. Shigella flexneri induces apoptosis in infected macrophages. Nature. 1992 Jul 9;358(6382):167–169. doi: 10.1038/358167a0. [DOI] [PubMed] [Google Scholar]

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