Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Sep;65(9):3615–3621. doi: 10.1128/iai.65.9.3615-3621.1997

Necrosis versus apoptosis as the mechanism of target cell death induced by Entamoeba histolytica.

O Berninghausen 1, M Leippe 1
PMCID: PMC175514  PMID: 9284127

Abstract

The human pathogen Entamoeba histolytica is known to kill a variety of host cells, including leukocytes. Using human myeloid cells as targets, we studied whether cytotoxicity of amoebic trophozoites in vitro is equivalent to the induction of apoptosis or whether these target cells die via necrosis. Based upon morphological criteria, incubation of target cells with amoebae resulted in necrosis, with cell swelling, rupture of plasma membrane, and release of cell contents including nucleic acids being detected by light and transmission electron microscopy. On the other hand, the characteristic features of apoptosis such as cell shrinking, surface blebbing, and chromatin condensation were not observed. Moreover, internucleosomal fragmentation of genomic DNA within target cells as a characteristic feature of apoptotic cell death did not occur as judged by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling technique in combination with flow cytometry. Consistently, cleavage of DNA was detectable upon agarose gel electrophoresis only after a substantial part of the target cell population had already been lysed. We also analyzed the mechanism of cell death induced by amoebapores, pore-forming peptides and primary candidate molecules for mediating the cytolytic activity of E. histolytica. At a time point at which the majority of target cells showed membrane injury upon incubation with purified amoebapores, no DNA degradation was detectable in the victim cells. The data suggest that the target cells used in our study undergo necrosis rather than apoptosis when they are killed by viable trophozoites as well as by isolated amoebapores.

Full Text

The Full Text of this article is available as a PDF (2.9 MB).

Selected References

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

  1. Ackers J. P. The invasiveness of Entamoeba histolytica - a continuing enigma. Clin Mol Pathol. 1996 Aug;49(4):M192–M198. doi: 10.1136/mp.49.4.m192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersson M., Gunne H., Agerberth B., Boman A., Bergman T., Sillard R., Jörnvall H., Mutt V., Olsson B., Wigzell H. NK-lysin, a novel effector peptide of cytotoxic T and NK cells. Structure and cDNA cloning of the porcine form, induction by interleukin 2, antibacterial and antitumour activity. EMBO J. 1995 Apr 18;14(8):1615–1625. doi: 10.1002/j.1460-2075.1995.tb07150.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bhakdi S., Weller U., Walev I., Martin E., Jonas D., Palmer M. A guide to the use of pore-forming toxins for controlled permeabilization of cell membranes. Med Microbiol Immunol. 1993 Sep;182(4):167–175. doi: 10.1007/BF00219946. [DOI] [PubMed] [Google Scholar]
  4. Diamond L. S., Harlow D. R., Cunnick C. C. A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg. 1978;72(4):431–432. doi: 10.1016/0035-9203(78)90144-x. [DOI] [PubMed] [Google Scholar]
  5. Ferrante A., Thong Y. H. Optimal conditions for simultaneous purification of mononuclear and polymorphonuclear leucocytes from human blood by the Hypaque-Ficoll method. J Immunol Methods. 1980;36(2):109–117. doi: 10.1016/0022-1759(80)90036-8. [DOI] [PubMed] [Google Scholar]
  6. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Guzmán C. A., Domann E., Rohde M., Bruder D., Darji A., Weiss S., Wehland J., Chakraborty T., Timmis K. N. Apoptosis of mouse dendritic cells is triggered by listeriolysin, the major virulence determinant of Listeria monocytogenes. Mol Microbiol. 1996 Apr;20(1):119–126. doi: 10.1111/j.1365-2958.1996.tb02494.x. [DOI] [PubMed] [Google Scholar]
  8. Jonas D., Walev I., Berger T., Liebetrau M., Palmer M., Bhakdi S. Novel path to apoptosis: small transmembrane pores created by staphylococcal alpha-toxin in T lymphocytes evoke internucleosomal DNA degradation. Infect Immun. 1994 Apr;62(4):1304–1312. doi: 10.1128/iai.62.4.1304-1312.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kolber M. A., Quinones R. R., Gress R. E., Henkart P. A. Measurement of cytotoxicity by target cell release and retention of the fluorescent dye bis-carboxyethyl-carboxyfluorescein (BCECF). J Immunol Methods. 1988 Apr 6;108(1-2):255–264. doi: 10.1016/0022-1759(88)90427-9. [DOI] [PubMed] [Google Scholar]
  10. Leippe M. Amoebapores. Parasitol Today. 1997 May;13(5):178–183. doi: 10.1016/s0169-4758(97)01038-7. [DOI] [PubMed] [Google Scholar]
  11. Leippe M. Ancient weapons: NK-lysin, is a mammalian homolog to pore-forming peptides of a protozoan parasite. Cell. 1995 Oct 6;83(1):17–18. doi: 10.1016/0092-8674(95)90229-5. [DOI] [PubMed] [Google Scholar]
  12. Leippe M., Andrä J., Nickel R., Tannich E., Müller-Eberhard H. J. Amoebapores, a family of membranolytic peptides from cytoplasmic granules of Entamoeba histolytica: isolation, primary structure, and pore formation in bacterial cytoplasmic membranes. Mol Microbiol. 1994 Dec;14(5):895–904. doi: 10.1111/j.1365-2958.1994.tb01325.x. [DOI] [PubMed] [Google Scholar]
  13. Leippe M., Müller-Eberhard H. J. The pore-forming peptide of Entamoeba histolytica, the protozoan parasite causing human amoebiasis. Toxicology. 1994 Feb 28;87(1-3):5–18. doi: 10.1016/0300-483x(94)90151-1. [DOI] [PubMed] [Google Scholar]
  14. Mangan D. F., Taichman N. S., Lally E. T., Wahl S. M. Lethal effects of Actinobacillus actinomycetemcomitans leukotoxin on human T lymphocytes. Infect Immun. 1991 Sep;59(9):3267–3272. doi: 10.1128/iai.59.9.3267-3272.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McCoy J. J., Mann B. J., Petri W. A., Jr Adherence and cytotoxicity of Entamoeba histolytica or how lectins let parasites stick around. Infect Immun. 1994 Aug;62(8):3045–3050. doi: 10.1128/iai.62.8.3045-3050.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Peitsch M. C., Polzar B., Stephan H., Crompton T., MacDonald H. R., Mannherz H. G., Tschopp J. Characterization of the endogenous deoxyribonuclease involved in nuclear DNA degradation during apoptosis (programmed cell death). EMBO J. 1993 Jan;12(1):371–377. doi: 10.1002/j.1460-2075.1993.tb05666.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Peña S. V., Hanson D. A., Carr B. A., Goralski T. J., Krensky A. M. Processing, subcellular localization, and function of 519 (granulysin), a human late T cell activation molecule with homology to small, lytic, granule proteins. J Immunol. 1997 Mar 15;158(6):2680–2688. [PubMed] [Google Scholar]
  18. Ragland B. D., Ashley L. S., Vaux D. L., Petri W. A., Jr Entamoeba histolytica: target cells killed by trophozoites undergo DNA fragmentation which is not blocked by Bcl-2. Exp Parasitol. 1994 Nov;79(3):460–467. doi: 10.1006/expr.1994.1107. [DOI] [PubMed] [Google Scholar]
  19. Ravdin J. I. Amebiasis. Clin Infect Dis. 1995 Jun;20(6):1453–1466. doi: 10.1093/clinids/20.6.1453. [DOI] [PubMed] [Google Scholar]
  20. Steiner H., Hultmark D., Engström A., Bennich H., Boman H. G. Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature. 1981 Jul 16;292(5820):246–248. doi: 10.1038/292246a0. [DOI] [PubMed] [Google Scholar]
  21. Steller H. Mechanisms and genes of cellular suicide. Science. 1995 Mar 10;267(5203):1445–1449. doi: 10.1126/science.7878463. [DOI] [PubMed] [Google Scholar]
  22. Vaux D. L. Toward an understanding of the molecular mechanisms of physiological cell death. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):786–789. doi: 10.1073/pnas.90.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Vermes I., Haanen C. Apoptosis and programmed cell death in health and disease. Adv Clin Chem. 1994;31:177–246. doi: 10.1016/s0065-2423(08)60336-4. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES