Interação entre Trypanosoma cruzi e macrófagos: diferenças entre tripomastigotas sangüí-colas e de cultivo de tecidos

Kloetzel, Judith K.; Milder, Regina V.; Umezawa, Eufrosina S.

doi:10.1590/S0036-46651984000400001

Resumos

Macrófagos obtidos do peritoneo de camundongos após estímulo, com peptona, foram cultivados em lamínulas, infectados com tripomastigotas das cepas F e Y de T. cruzi, obtidos de cultivo de tecidos ou do sangue de camundongos infectados. Os parasitas, obtidos de cultivo de tecidos, tanto da cepa Y como os da cepa F, são interiorizados por macrófagos em proporção muito mais elevada do que os sanguícolas. Parasitas de cultivo de tecidos incubados com soro de camundongos normais, ou soro híperimune específico em diluição sub-aglutinante, comportam-se essencialmente como parasitas não opsonizados. Foram observadas diferenças a nível ultraestrutural na fase inicial de interação entre macrófagos e tripomastigotas das duas origens. Após 30 minutos, tripomastigotas de cultivo de tecidos localizam-se em agrupamentos na área de contato com os macrófagos. Enquanto os tripomastigotas sanguícolas estão na maioria das vezes no interior de vacúolos fagocíticos largos, após 3 horas de interação os tripomastigotas de cultura situam-se em um único vacúolo estreito. Tanto as formas de cultivo de tecidos quanto os tripomastigotas sanguícolas da cepa Y multiplicam-se em macrófagos; os tripomastigotas sanguícolas da cepa F são destruídos no interior da célula hospedeira, enquanto os tripomastigotas de cultivo de tecidos desta cepa são capazes de multiplicar-se.

Mouse peritoneal elicited macrophages cultured on coverslips were infected with Trypanosoma cruzi trypomastigotes from both the F strain and Y strain obtained either from tissue culture or from the bloodstream of infected mice. Both the Y strain and F parasites obtained from tissue culture were interiorized by macrophages at a much higher rate than bloodstream trypomastigotes. Tissue culture parasites incubated with normal mouse serum, mouse plasma obtained at the 7th day after infection, or specific hyperimmune serum at subagglutinating concentration, behaved essentially as non-opsonized parasites. Ultrastructural differences were seen at the early interaction phase between macrophages and trypimastigotes from both sources. After 30 minutes, tissue culture trypomastigotes were located in clusters at the area of contact with macrophages. While bloodstream trypomastigotes, at 3 hours post-infection were most frequently enclosed in a loose phagocytic vacuole, tissue culture trypomastigotes were enclosed in single tight vacuoles. Both tissue culture and bloodstream trypomastigotes of the Y strain multiplied within macrophages; F strain bloodstream trypomastigotes did not develop within the host cells, while tissue culture trypomastigotes multiplied.

Trypanosoma cruzi interaction with macrophages: differences between tissue culture and bloodstream forms^* (* ) This work was supported by CNPq Grants 2222.8.097/80 and 2222.8.141/80 and FINEP Grant B7680/23/00/00 Address: Av. Dr. Enéias C Aguiar 470, 05403 São Paulo, Brasil

Interação entre Trypanosoma cruzi e macrófagos: diferenças entre tripomastigotas sangüí-colas e de cultivo de tecidos

Judith K. Kloetzel^I; Regina V. Milder^II; Eufrosina S. Umezawa^I

^IInstituto de Medicina Tropical de São Paulo, University of São Paulo School of Medicine, Department of Preventive Medicine

^IIInstituto de Medicina Tropical de São Paulo, University of São Paulo School of Medicine, Department of Parasitology

SUMMARY

Mouse peritoneal elicited macrophages cultured on coverslips were infected with Trypanosoma cruzi trypomastigotes from both the F strain and Y strain obtained either from tissue culture or from the bloodstream of infected mice. Both the Y strain and F parasites obtained from tissue culture were interiorized by macrophages at a much higher rate than bloodstream trypomastigotes. Tissue culture parasites incubated with normal mouse serum, mouse plasma obtained at the 7th day after infection, or specific hyperimmune serum at subagglutinating concentration, behaved essentially as non-opsonized parasites. Ultrastructural differences were seen at the early interaction phase between macrophages and trypimastigotes from both sources. After 30 minutes, tissue culture trypomastigotes were located in clusters at the area of contact with macrophages. While bloodstream trypomastigotes, at 3 hours post-infection were most frequently enclosed in a loose phagocytic vacuole, tissue culture trypomastigotes were enclosed in single tight vacuoles. Both tissue culture and bloodstream trypomastigotes of the Y strain multiplied within macrophages; F strain bloodstream trypomastigotes did not develop within the host cells, while tissue culture trypomastigotes multiplied.

RESUMO

Macrófagos obtidos do peritoneo de camundongos após estímulo, com peptona, foram cultivados em lamínulas, infectados com tripomastigotas das cepas F e Y de T. cruzi, obtidos de cultivo de tecidos ou do sangue de camundongos infectados. Os parasitas, obtidos de cultivo de tecidos, tanto da cepa Y como os da cepa F, são interiorizados por macrófagos em proporção muito mais elevada do que os sanguícolas. Parasitas de cultivo de tecidos incubados com soro de camundongos normais, ou soro híperimune específico em diluição sub-aglutinante, comportam-se essencialmente como parasitas não opsonizados. Foram observadas diferenças a nível ultraestrutural na fase inicial de interação entre macrófagos e tripomastigotas das duas origens. Após 30 minutos, tripomastigotas de cultivo de tecidos localizam-se em agrupamentos na área de contato com os macrófagos. Enquanto os tripomastigotas sanguícolas estão na maioria das vezes no interior de vacúolos fagocíticos largos, após 3 horas de interação os tripomastigotas de cultura situam-se em um único vacúolo estreito. Tanto as formas de cultivo de tecidos quanto os tripomastigotas sanguícolas da cepa Y multiplicam-se em macrófagos; os tripomastigotas sanguícolas da cepa F são destruídos no interior da célula hospedeira, enquanto os tripomastigotas de cultivo de tecidos desta cepa são capazes de multiplicar-se.

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1. ALCANTARA, A. & BRENER, Z. Trypanosoma cruzi: role of macrophage membrane components in the phagocytosis of bloodstream forms. Exp. Parasit. 50: 1-6, 1980.
2. BRENER, Z. Immunity to Trypanosoma cruzi Advanc. Parasit. 18: 247-292, 1980.
3. CHANG, K. P. Leishmania donovani: promastigote-macrophage surface interaction in vitro. Exp. Parasit. 48: 175-189, 1979.
4. DEANE, M. P. & KLOETZEL, J. Lack of protection against Trypanosoma cruzi by multiple doses of T. lewisi culture forms. A discussion on some strains of "lewisi". Exp. Parasit. 35: 406-410, 1974.
5. DVORAK, J. A. & SCHMUNIS, G. A. Trypanosoma cruzi: interaction with mouse peritoneal macrophages. Exp. Parasit. 23: 289-3G0, 1972.
6. KIPNIS, T. L.; DAVID, J. R.; ALPER, C. A.; SHER, A. & DIAS DA SILVA, W. Enzymatic treatment transforms trypomastigotes of Trypanosoma cruzi into activators of alternative complement pathway and potentiates their uptake by macrophages. Proc. Natl. Acad. Sc. USA 78: 602-605, 1981.
7. KLOETZEL, J. & DEANE, M. P. Presence of immunoglobulins on the surface of bloodstream Trypanosoma cruzi Capping during differentiation in culture. Rev. Inst. Med. trop. São Paulo 19: 397-402, 1977.
8. KRETTLI, A. U. & NUSSENZWEIG, R. S. Presence of immunoglobulin on the surface of circulating trypomastigotes of T. cruzi resulting in activation of the alternative pathway of complement and lysis. Washington, P.A.H.O. Sci. Publ. 347: 71-73, 1977.
9. MARIA, T. A.; ALCANTARA, A. & BRENER, Z. Ultrastructural studies on the in vitro interaction of Trypanosoma cruzi bloodstream forms and mouse peritoneal macrophages. Acta trop. 39: 99-109, 1982.
10. MEIRELLES, M. N. L.; CHIARI, E. & SOUZA, W. Interaction of bloodstream, tissue culture-derived and axenic-culture-derived trypomastigotes of Trypanosoma cruzi with macrophages. Acta trop. 39: 195-203, 1982.
11. MILDER, R. & KLOETZEL, J. The development of Trypanosoma cruzi in macrophages in vitro. Interaction with lysosomes and host cell fate. Parasitology 80: 139-145, 1980.
12. MILDER, R.; KLOETZEL, J. & DEANE, M. Observation on the interaction of peritoneal macrophages with Trypanosoma cruzi II. Intracellular fate of bloodstream forms. Rev. Inst. Med. trop. São Paulo 19: 313-322, 1977.
13. MIRANDA-SANTOS, I. K. Ferreira de & CAMPOS NETO, A. Receptors for immunoglobulin Fc on pathogenic but not on nonpathogenic Protozoa of the Trypanosomatidae. J. Exp. Med. 154: 1732-1742, 1981.
14. NOGUEIRA, N.; CHAPLAN, S. & COHN, Z. Trypanosoma cruzi Factors modifying ingestion and fate of blood form trypomastigotes. J. Exp. Med. 152: 447-451, 1980.
15. SANDERSON, C. J. & SOUZA, W. de A morphological study of the interaction between Trypanosoma cruzi and eosinophils, neutrophils and macrophages in vitro. J. Cell Sci. 37: 275-286, 1979.
16. SILVA, L. H. P. da & NUSSENZWEIG, V. Sobre uma cepa de T. cruzi altamente virulenta para o camundongo branco. Folia Clin. et Biol. (S. Paulo) 20: 191-208, 1953.
17. TANOWITZ, H. : WITTNER, M.; KRESS, Y. & BLOOM, B. Studies of the in vitro infection by Trypanosoma cruzi I. Ultrastructural studies on the invasion of macrophages and L-cells. Amer. J. trop. Med. Hyg. 24: 25-33, 1975.

(*

) This work was supported by CNPq Grants 2222.8.097/80 and 2222.8.141/80 and FINEP Grant B7680/23/00/00 Address: Av. Dr. Enéias C Aguiar 470, 05403 São Paulo, Brasil

Datas de Publicação

Publicação nesta coleção
08 Abr 2013
Data do Fascículo
Ago 1984

Histórico

Recebido
09 Nov 1983

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. ALCANTARA, A. & BRENER, Z. Trypanosoma cruzi: role of macrophage membrane components in the phagocytosis of bloodstream forms. Exp. Parasit. 50: 1-6, 1980.

[2] 2. BRENER, Z. Immunity to Trypanosoma cruzi Advanc. Parasit. 18: 247-292, 1980.

[3] 3. CHANG, K. P. Leishmania donovani: promastigote-macrophage surface interaction in vitro. Exp. Parasit. 48: 175-189, 1979.

[4] 4. DEANE, M. P. & KLOETZEL, J. Lack of protection against Trypanosoma cruzi by multiple doses of T. lewisi culture forms. A discussion on some strains of "lewisi". Exp. Parasit. 35: 406-410, 1974.

[5] 5. DVORAK, J. A. & SCHMUNIS, G. A. Trypanosoma cruzi: interaction with mouse peritoneal macrophages. Exp. Parasit. 23: 289-3G0, 1972.

[6] 6. KIPNIS, T. L.; DAVID, J. R.; ALPER, C. A.; SHER, A. & DIAS DA SILVA, W. Enzymatic treatment transforms trypomastigotes of Trypanosoma cruzi into activators of alternative complement pathway and potentiates their uptake by macrophages. Proc. Natl. Acad. Sc. USA 78: 602-605, 1981.

[7] 7. KLOETZEL, J. & DEANE, M. P. Presence of immunoglobulins on the surface of bloodstream Trypanosoma cruzi Capping during differentiation in culture. Rev. Inst. Med. trop. São Paulo 19: 397-402, 1977.

[8] 8. KRETTLI, A. U. & NUSSENZWEIG, R. S. Presence of immunoglobulin on the surface of circulating trypomastigotes of T. cruzi resulting in activation of the alternative pathway of complement and lysis. Washington, P.A.H.O. Sci. Publ. 347: 71-73, 1977.

[9] 9. MARIA, T. A.; ALCANTARA, A. & BRENER, Z. Ultrastructural studies on the in vitro interaction of Trypanosoma cruzi bloodstream forms and mouse peritoneal macrophages. Acta trop. 39: 99-109, 1982.

[10] 10. MEIRELLES, M. N. L.; CHIARI, E. & SOUZA, W. Interaction of bloodstream, tissue culture-derived and axenic-culture-derived trypomastigotes of Trypanosoma cruzi with macrophages. Acta trop. 39: 195-203, 1982.

[11] 11. MILDER, R. & KLOETZEL, J. The development of Trypanosoma cruzi in macrophages in vitro. Interaction with lysosomes and host cell fate. Parasitology 80: 139-145, 1980.

[12] 12. MILDER, R.; KLOETZEL, J. & DEANE, M. Observation on the interaction of peritoneal macrophages with Trypanosoma cruzi II. Intracellular fate of bloodstream forms. Rev. Inst. Med. trop. São Paulo 19: 313-322, 1977.

[13] 13. MIRANDA-SANTOS, I. K. Ferreira de & CAMPOS NETO, A. Receptors for immunoglobulin Fc on pathogenic but not on nonpathogenic Protozoa of the Trypanosomatidae. J. Exp. Med. 154: 1732-1742, 1981.

[14] 14. NOGUEIRA, N.; CHAPLAN, S. & COHN, Z. Trypanosoma cruzi Factors modifying ingestion and fate of blood form trypomastigotes. J. Exp. Med. 152: 447-451, 1980.

[15] 15. SANDERSON, C. J. & SOUZA, W. de A morphological study of the interaction between Trypanosoma cruzi and eosinophils, neutrophils and macrophages in vitro. J. Cell Sci. 37: 275-286, 1979.

[16] 16. SILVA, L. H. P. da & NUSSENZWEIG, V. Sobre uma cepa de T. cruzi altamente virulenta para o camundongo branco. Folia Clin. et Biol. (S. Paulo) 20: 191-208, 1953.

[17] 17. TANOWITZ, H. : WITTNER, M.; KRESS, Y. & BLOOM, B. Studies of the in vitro infection by Trypanosoma cruzi I. Ultrastructural studies on the invasion of macrophages and L-cells. Amer. J. trop. Med. Hyg. 24: 25-33, 1975.