Cell and Tissue Research

, Volume 314, Issue 2, pp 223–235 | Cite as

Host and parasite apoptosis following Trypanosoma cruzi infection in in vitro and in vivo models

  • E. M. de Souza
  • T. C. Araújo-Jorge
  • C. Bailly
  • A. Lansiaux
  • M. M. Batista
  • G. M. Oliveira
  • M. N. C. Soeiro
Regular Article

Abstract

The mechanism of cell death which occurs during Chagas' cardiopathy is disputed. To address this issue we analyzed the molecular pathways implicated in the death of cardiomyocytes during T. cruzi invasion and found that they undergo apoptosis during both in vitro and in vivo infections. However, the death rates and onset were related to the parasite stocks belonging to different biodemes, which can be correlated to the different histological inflammation findings that have already been reported. Our in vitro data provide additional support for this hypothesis since higher levels and earlier apoptosis induction were noted during the interaction with the Dm28c (type I) as compared to the Y and CL stocks (type II). Modifications of the surface carbohydrates of the infected cardiomyocytes were observed and these molecular events may be acting as "eat me" tags for their final engulfment by macrophages and/or other non-professional phagocytes. The analysis of other host cell types showed that the in vitro infection of fibroblasts did not result in host apoptosis even when a highly infective stock was used. Conversely, infected macrophages undergo apoptosis but at a higher degree than cardiomyocytes. Apoptotic intracellular parasites were observed to varied extents depending on the T. cruzi stock, which was related to the parasite invasion and proliferation. In summary, our results show that during T. cruzi infection, the extent of apoptosis varies according to the host cell type and the parasite stocks. The apoptosis of both host and T. cruzi can contribute to the silent spreading and persistence of the parasite without triggering an exacerbated inflammatory response.

Keywords

Cardiomyocytes Biodemes Apoptosis Trypanosoma cruzi (Protozoa, Flagellata) 

Notes

Acknowledgements

We thank Bruno Ávila for excellent work on the image processing.

References

  1. Al-Olayan EM, Williams GT, Hurd H (2002) Apoptosis in the malaria protozoan, Plasmodium berghei: a possible mechanism for limiting intensity of infection in the mosquito. J Parasitol 32:1133–1143CrossRefGoogle Scholar
  2. Andrade SG, Magalhães JB (1996) Biodemes and zymodemes of Trypanosoma cruzi strains: correlations with clinical data and experimental pathology. Rev Soc Bras Med Trop 30:27–35PubMedGoogle Scholar
  3. Araújo Jorge TC, Barbosa HS, Moreira AL, De Souza W, Meirelles MN (1986) The interaction of myotropic and macrophagotropic strains of Trypanosoma cruzi with myoblasts and fibers of skeletal muscle. Z Parasitenkd 72:577–584PubMedGoogle Scholar
  4. Araújo-Jorge TC, Sampaio ED, De Souza W, Meirelles MN (1989) T. cruzi: the effect of variations in experimental conditions on the levels of macrophage infection "in vitro". Parasitol Res 75:257–263PubMedGoogle Scholar
  5. Arnoult D, Akarid K, Grodet A, Petit PX, Estaquier J, Ameisen JC (2002) On the evolution of programmed cell death: apoptosis of the unicellular eukaryote Leishmania major involves cysteine proteinase activation and mitochondrion permeabilization. Cell Death Differ 9:65–81CrossRefPubMedGoogle Scholar
  6. Behnia M, Robertson KA, Martin WJ (2000) Role of apoptosis in host defense and pathogenesis of disease. Chest 117:1771–1777CrossRefPubMedGoogle Scholar
  7. Chuenkova MV, Pereira MA (2000) A trypanosomal protein synergizes with the cytokines ciliary neurotrophic factor and leukemia inhibitory factor to prevent apoptosis of neuronal cells. Mol Biol Cell 11:1487–1498PubMedGoogle Scholar
  8. Clark RK, Kuhn RE (1999) Trypanosoma cruzi does not induce apoptosis in murine fibroblasts. Parasitology 118:167–175CrossRefPubMedGoogle Scholar
  9. Contreras VT, Araújo-Jorge TC, Bonaldo MC, Thomaz N, Barbosa HS, Meirelles MN, Goldenberg S (1988) Biological aspects of the Dm28c clone of Trypanosoma cruzi after metacyclogenesis in chemically defined media. Mem Inst Oswaldo Cruz 83:123–133PubMedGoogle Scholar
  10. Corte-Real S, Santos CB, Meirelles MN (1995) Differential expression of the plasma membrane Mg2+ ATPase and Ca2+ ATPase activity during adhesion and interiorization of Leishmania amazonensis in fibroblasts in vitro. J Submicrosc Cytol Pathol 27:359–366Google Scholar
  11. Das G, Vohra H, Saha B, Agrewala JN, Mishra GC (1998) Leismania donovani infection of a susceptible host results in apoptosis of Th1-like cells: rescue of anti-leishmanial CMI by providing Th1-specific bystander costimulation. Microbiol Immunol 42:795–801PubMedGoogle Scholar
  12. Dini L, Pagliara P, Carla EC (2002) Phagocytosis of apoptotic cells by liver: a morphological study. Microsc Res Tech 57:530–540CrossRefPubMedGoogle Scholar
  13. Frasch AC (1994) Trans-sialidase, SAPA amino acid repeats and the relationship between Trypanosoma cruzi and the mammalian host. Parasitology 108:37–44Google Scholar
  14. Freire-de-Lima CG, Nunes MP, Corte-Leal S, Soares MP, Previato JO, Mendonça-Previato, DosReis GA (1998) Proapoptotic activity of a Trypanosoma cruzi ceramida-containing glycolipid turned on in host macrophages by IFN-g. J Immunol 161:4909–4916PubMedGoogle Scholar
  15. Freire-de-Lima CG, Nascimento DO, Soares MB, Bozza PT, Castro-Faria-Neto HC, de Mello FG, Dos Reis GA, Lopes MF (2000) Uptake of apoptotic cells drives the growth of a pathogenic trypanosome in macrophages. Nature 403:199–203CrossRefPubMedGoogle Scholar
  16. Goebel S, Luder CG, Lugert R, Bohne W, Gross U (1998) Toxoplasma gondii inhibits the in vitro induced apoptosis of HL-60 cells. J Exp Clin Med 23:351–356Google Scholar
  17. Goebel S, Luder CG, Gross U (1999) Invasion by Toxoplasma gondii protects human-derived HL-60 cell from actinomycin D-induced apoptosis. Med Microbiol Immunol 187:221–226CrossRefPubMedGoogle Scholar
  18. Henriques-Pons A, Oliveira GM, Paiva MM, Correa AF, Batista MM, Bisaggio RC, Liu CC, Cotta-De-Almeida V, Coutinho CM, Persechini PM, Araújo-Jorge TC (2002) Evidence for a perforin-mediated mechanism controlling cardiac inflammation in Trypanosoma cruzi infection. Int J Exp Pathol 83:67–79CrossRefPubMedGoogle Scholar
  19. Heussler VT, Kuenzi P, Rottenberg S (2001) Inhibition of apoptosis by intracellular protozoan parasites. Int J Parasitol 31:1166–1176CrossRefPubMedGoogle Scholar
  20. Kanoh M, Takemura G, Misao J, Hayakawa Y, Aoyama T, Nishigaki K, Noda T, Fujiwara T, Fukuda K, Minatoguchi S, Fujiwara H (1999) Significance of myocytes with positive DNA in situ nick end-labelling (TUNEL) in hearts with dilated cardiomyopathy: not apoptosis but DNA repair. Circulation 99:2757–2764PubMedGoogle Scholar
  21. Kayakawa K, Takemura G, Koda M, Kawase Y, Maruyama R, Li Y, Minatigushi S, Fujiwara H (2002) Sensitivity to apoptosis signal, clearance rate, and ultrastructural of Fas ligand-induced apoptosis in in vivo adult cardiac cells. Circulation 105:3039–3045CrossRefPubMedGoogle Scholar
  22. Keane J, Remold HG, Kornfeld H (2000) Virulent Mycobacterium tuberculosis strains evade apoptosis of infected alveolar macrophages. J Immunol 164:2016–2020PubMedGoogle Scholar
  23. Khan IA, Matsuura T, Kasper LH (1996) Activation-mediated CD4+ T cell unresponsiveness during acute Toxoplasma gondii infection in mice. Int Immunol 8:887–8896PubMedGoogle Scholar
  24. Kluza J, Lansiaux A, Wattez M, Mahieu C, Osheroff N, Bailly C (2000) Apoptotic response of HL-60 human leukemia cells to the antitumor drug TAS-103. Cancer Res 60:4077–4084PubMedGoogle Scholar
  25. Lee N, Bertholet S, Debrabant A, Muller J, Duncan R, Nakhasi HL (2002) Programmed cell death in the unicellular protozoan parasite Leishmania. Cell Death Differ 9:53–64CrossRefPubMedGoogle Scholar
  26. Leguizamon MS, Mocetti E, Garcia Rivello H, Argibay P, Campetella O (1999) Trans-sialidase from Trypanosoma cruzi induces apoptosis in cells from the immune system in vivo. J Infect Dis 180:1398–1402CrossRefPubMedGoogle Scholar
  27. Lopes MF, Da Veiga VF, Santos AR, Fonseca ME, DosReis GA (1995a) Activation-induced CD4+ T cell death by apoptosis in experimental Chagas' disease. J Immunol 154:744–752PubMedGoogle Scholar
  28. Lopes MF, Cunha JM, Bezerra FL, Gonzalez MS, Gomes JE, Lapa e Silva JR, Garcia ES, DosReis GA (1995b) Trypanosoma cruzi: both chemically induced and triatomine-derived metacyclic trypomastigotes cause the same immunological disturbances in the infected mammalian host. Exp Parasitol 80:194–204CrossRefPubMedGoogle Scholar
  29. Lopes MF, Nunes MP, Henriques-Pons A, Giese N, Morse HC III, Davison WF, Araújo-Jorge TC, DosReis GA (1999) Increased susceptibility of Fas ligand-deficient gld mice to Trypanosoma cruzi infection due to a Th2-biased host immune response. Eur J Immunol 29:81–89CrossRefPubMedGoogle Scholar
  30. Lüder CG, Gross U, Lopes MF (2001) Intracellular protozoan parasites and apoptosis: diverse strategies to modulate parasite-host interactions. Trends Parasitol 17:480–486CrossRefPubMedGoogle Scholar
  31. Machado FS, Martins GA, Aliberti JC, Mestriner FL, Cunha FQ, Silva JS (2000) Trypanosoma cruzi-infected cardiomyocytes produce chemokines and cytokines that trigger potent nitric oxide-dependent trypanocidal activity. Circulation 102:3003–3008PubMedGoogle Scholar
  32. Meirelles MNL, Araújo-Jorge TC, De Souza W (1982) Interaction of Trypanosoma cruzi with macrophages in vitro: dissociation of the attachment and internalization phases by low temperature and cytochalasin B. Z Parasitenkd 68:7–14PubMedGoogle Scholar
  33. Meirelles MNL, Araújo-Jorge TC, Miranda CF, De Souza W, Barbosa HS (1986) Interaction of Trypanosoma cruzi with heart muscle cells: ultrastructural and cytochemical analysis of endocytic vacuole formation and effect upon myogenesis in in vitro. Eur J Cell Biol 41:198–206PubMedGoogle Scholar
  34. Meirelles MN, Pereira MC, Singer RH, Soeiro MN, Garzoni LR, Silva DT, Barbosa HS, Araújo-Jorge TC, Masuda MO, Capella MA, Lopes AG, Vermelho AB (1999) Trypanosoma cruzi-cardiomyocytes: new contributions regarding a better understanding of this interaction. Mem Inst Oswaldo Cruz 94 Suppl 1:149–152Google Scholar
  35. Morris RG, Hargreaves AD, Duvall E, Wyllie AH (1984) Hormone-induced cell death. 2. Surface changes in thymocytes undergoing apoptosis. Am J Pathol 115:426–436PubMedGoogle Scholar
  36. Mucci J, Hidalgo A, Mocetti E, Argibay PF, Leguizamon MS, Campetella O (2002) Thymocyte depletion in Trypanosoma cruzi infection is mediated by trans-sialidase-induced apoptosis on nurse cells complex. Proc Natl Acad Sci U S A 99:3896–3901CrossRefPubMedGoogle Scholar
  37. Nakajiima-Shimada J, Zou C, Takagi M, Umeda M, Nara T, Aoki T (2000) Inhibition of Fas-mediated apoptosis by Trypanosoma cruzi infection. Biochim Biophys Acta 1475:175–183PubMedGoogle Scholar
  38. Palomino SA, Aiello VD, Higuchi ML (2000) Systematic mapping of hearts from chronic chagasic patients: the association between the occurrence of histopathological lesions and Trypanosoma cruzi antigens. Ann Trop Med Parasitol 94:571–579PubMedGoogle Scholar
  39. Pereira MC, Costa M, Chagas Filho C, Meirelles MN (1993) Myofibrillar breakdown and cytoskeletic alterations in heart muscle cells during invasion by Trypanosoma cruzi: immunological and ultrastructural study. J Submicrosc Cytol Pathol 25:559–569Google Scholar
  40. Pereira MC, Singer RH, Meirelles MN (2000) Trypanosoma cruzi infection affects actin mRNA regulation in heart muscle cells. J Eukaryot Microbiol 47:271–279PubMedGoogle Scholar
  41. Piacenza L, Peluffo G, Radi R (2001) l-Arginine-dependent suppression of apoptosis in Trypanosoma cruzi: contribution of the nitric oxide and polyamine pathways. Proc Natl Acad Sci U S A 98:7301–7306CrossRefPubMedGoogle Scholar
  42. Platt N, da Silva RP, Gordon S (1998) Class a scavenger receptors and the phagocytosis of apoptotic cells. Biochem Soc Trans 26:639–644PubMedGoogle Scholar
  43. Quan N, Mhlanga JD, Whiteside MB, McCoy AN, Kristensson K, Herkenham M (1999) Chronic over expression of proinflammatory cytokines and histopathology in the brains of rats infected with Trypanosoma brucei. J Comp Neurol 414:114–130CrossRefPubMedGoogle Scholar
  44. Roggero E, Perez A, Tamae-Kakazu M, Piazzon I, Nepomnaschy I, Wietzerbin J, Serra E, Revelli S, Bottasso O (2002) Differential susceptibility to acute Trypanosoma cruzi infection in BALB/c and C57BL/6 mice is not associated with a distinct parasite load but cytokine abnormalities. Clin Exp Immunol 128:421–428CrossRefPubMedGoogle Scholar
  45. Schenkman S, Jiang MS, Hart GW, Nussenzweig V (1991) A novel cell surface trans-sialidase of Trypanosoma cruzi generates a stage-specific epitope required for invasion of mammalian cells. Cell 65:1117–1125PubMedGoogle Scholar
  46. Soeiro MNC, Silva-Filho FC, Meirelles MNL (1995) Alterations in the surface charge of heart muscle cells during interaction with T. cruzi. Cell Biophysics 26:21–44PubMedGoogle Scholar
  47. Soeiro MNC, Paiva MM, Barbosa HS, Meirelles MNL, Araújo-Jorge TC (1999) A cardiomyocyte mannose receptor system is involved in Trypanosoma cruzi invasion and is down modulated after infection. Cell Struct Funct 24:139–149CrossRefPubMedGoogle Scholar
  48. Soeiro MNC, Mota RA, Batista DGJ, Pereira MCS, Meirelles MNL (2002) Trypanosoma cruzi infection impairs zymosam uptake by cardiomyocytes. Pathobiology 70:69–75CrossRefPubMedGoogle Scholar
  49. Timms MW, van Deursen FJ, Hendriks EF, Matthews KR (2002) Mitochondrial development during life cycle differentiation of African trypanosomes: evidence for a kinetoplast-dependent differentiation control point. Mol Biol Cell 13:3747–3759CrossRefPubMedGoogle Scholar
  50. Ullrich CK, Groopman JE, Ganju RK (2000) HIV-1 gp 120 and gp 160-induced apoptosis in cultured endothelial cells is mediated by caspases. Blood 96:1438–1442PubMedGoogle Scholar
  51. Waghabi MC, Coutinho CM, Soeiro MN, Pereira MC, Feige JJ, Keramidas M, Cosson A, Minoprio P, Van Leuven F, Araújo-Jorge TC (2002) Increased Trypanosoma cruzi invasion and heart fibrosis associated with high transforming growth factor beta levels in mice deficient in alpha (2)-macroglobulin. Infect Immun 70:5115–5123CrossRefPubMedGoogle Scholar
  52. Weinrauch Y, Zychlinsky A (1999) The induction of apoptosis by bacterial pathogens. Annu Rev Microbiol 53:155–187CrossRefPubMedGoogle Scholar
  53. WHO (1997) Chagas' disease: interruption of transmission. Weekly Epidemiol Rec 72:1–5Google Scholar
  54. Zhang J, Andrade ZA, Yu ZX, Andrade SG, Takeda K, Sadirgursky M, Ferrans VJ (1999) Apoptosis in a canine model of acute Chagasic myocarditis. J Mol Cell Cardiol 31:581–596CrossRefPubMedGoogle Scholar
  55. Zingales B, Pereira ME, Almeida KA, Umezawa ES, Nehme NS, Oliveira RP, Macedo A, Souto RP (1997) Biological parameters and molecular markers of clone CL Brener—the reference organism of the Trypanosoma cruzi genome project. Mem Inst Oswaldo Cruz 92:811–814PubMedGoogle Scholar
  56. Zingales B, Stolf BS, Souto RP, Fernandes O, Briones MR (1999) Epidemiology, biochemistry and evolution of Trypanosoma cruzi lineages based on ribosomal RNA sequences. Mem Inst Oswaldo Cruz 94 Suppl 1:159–164Google Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • E. M. de Souza
    • 1
  • T. C. Araújo-Jorge
    • 1
  • C. Bailly
    • 2
  • A. Lansiaux
    • 2
  • M. M. Batista
    • 1
  • G. M. Oliveira
    • 1
  • M. N. C. Soeiro
    • 1
  1. 1.Lab. Biologia Celular, DUBC, FIOCRUZInstituto Oswaldo CruzRio de JaneiroBrazil
  2. 2.INSERM U-524 Centre Oscar LambretLilleFrance

Personalised recommendations