Trypanosoma cruzi: modification of alkaline phosphatase activity induced by trypomastigotes in cultured human placental villi

Fretes, Ricardo E.; Fabro, Sofía P. de

doi:10.1590/S0036-46651990000600003

Abstracts

Human term placental villi cultured ''in vitro" were maintained with bloodstream forms of Trypanosoma cruzi during various periods of time. Two different concentrations of the parasite were employed. Controls contained no T. cruzi. The alkaline phosphatase activity was determined in placental villi by electron microscopy and its specific activity in the culture medium by biochemical methods. Results showed that the hemoflagellate produces a significant decrease in enzyme activity as shown by both ultracytochemical and specific activity studies and this activity was lower in cultures with high doses of parasites. The above results indicate that the reduction in enzyme activity coincides with the time of penetration and proliferation of T. cruzi in mammalian cells. These changes may represent an interaction between human trophoblast and T. cruzi.

Trypanosoma cruzi; Placental villi; Tissue coculture; Alkaline phosphatase; Chagas' disease

Vilos placentários humanos a termo foram mantidos "in vitro" em interação com formas tripomastigotas sangüíneas de Trypanosoma cruzi, durante diversos períodos de tempo. Foram utilizadas concentrações diferentes de parasitas. Os controles não continham T. cruzi. Determinou-se a atividade de fosfatase alcalina em vilos placentários mediante microscopia eletrônica e sua atividade específica no meio de cultura, mediante métodos bioquímicos. Os resultados mostraram que o hemoflagelado produz uma diminuição significante da atividade enzimática tanto pelos estudos ultracitoquímicos como de atividade específica e esta atividade de fosfatase alcalina foi menor em culturas com altas doses de parasitas. Estes resultados são indicadores de que a redução de atividade enzimática coincide com o tempo de penetração e proliferação do T. cruzi nas células. Estas mudanças podem representar uma interação entre o trofoblasto humano e o T. cruzi.

ORIGINAL ARTICLES

Trypanosoma cruzi: modification of alkaline phosphatase activity induced by trypomastigotes in cultured human placental villi

Trypanosoma cruzi: alteração da atividade de fosfatase alcalina induzida por tripomastigotas em culturas de vilos placentários humanos

Ricardo E. Fretes; Sofía P. de Fabro

II Cátedra de Histología y Embriología. Facultad de Ciencias Médicas. Universidad Nacional de Córdoba CONICET, Argentina

^{Address for correspondence} Address for correspondence: Dra. Sofía P. de Fabro La Noria, 1262 (5000) Córdoba, Argentina

SUMMARY

Human term placental villi cultured ''in vitro" were maintained with bloodstream forms of Trypanosoma cruzi during various periods of time. Two different concentrations of the parasite were employed. Controls contained no T. cruzi. The alkaline phosphatase activity was determined in placental villi by electron microscopy and its specific activity in the culture medium by biochemical methods. Results showed that the hemoflagellate produces a significant decrease in enzyme activity as shown by both ultracytochemical and specific activity studies and this activity was lower in cultures with high doses of parasites. The above results indicate that the reduction in enzyme activity coincides with the time of penetration and proliferation of T. cruzi in mammalian cells. These changes may represent an interaction between human trophoblast and T. cruzi.

Key words: Trypanosoma cruzi; Placental villi, human; Tissue coculture; Alkaline phosphatase; Chagas' disease, congenital.

RESUMO

Vilos placentários humanos a termo foram mantidos "in vitro" em interação com formas tripomastigotas sangüíneas de Trypanosoma cruzi, durante diversos períodos de tempo. Foram utilizadas concentrações diferentes de parasitas. Os controles não continham T. cruzi. Determinou-se a atividade de fosfatase alcalina em vilos placentários mediante microscopia eletrônica e sua atividade específica no meio de cultura, mediante métodos bioquímicos. Os resultados mostraram que o hemoflagelado produz uma diminuição significante da atividade enzimática tanto pelos estudos ultracitoquímicos como de atividade específica e esta atividade de fosfatase alcalina foi menor em culturas com altas doses de parasitas. Estes resultados são indicadores de que a redução de atividade enzimática coincide com o tempo de penetração e proliferação do T. cruzi nas células. Estas mudanças podem representar uma interação entre o trofoblasto humano e o T. cruzi.

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ACKNOWLEDGMENTS

We are grateful to Drs. Patricia Paglini and Adriana Gruppi for supplying trypomastigotes of Trypanosoma cruzi employed in this study We thank Dr. Gonzalez Cremer for critical reading of the manuscript. R. E. Fretes is a fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina (CONICET). This work was supported by grant 15732/88 from the Subsecretaría de Ciencia y Técnica de la Nación, Argentina.

Recebido para publicação em 16/2/1990.

1. ADENIYI, F.A. & OLATUNBOSUN, D.A. Origins and significance of the increased plasma alkaline phosphatase during normal pregnancy and pre-eclampsia. Brit. J. Obstet. Gynaec., 91: 857-862, 1984.
2. ANDRADE, S.G. The influence of the strain of Trypanosoma cruzi in placental infections in mice. Trans. roy. Soc. trop. Med. Hyg., 76: 123-128, 1982.
3. ANDREWS, N.W. & COLLI, W. Adhesion and interiorization of Trypanosoma cruzi in mammalian cells. J. Protozool., 29: 264-269, 1982.
4. BITTENCOURT, A.L. Congenital Chagas' disease. Amer. J. Dis. Child., 130: 97-103, 1976.
5. BONGERSZ, V. & HUNGERER, K.D. Trypanosoma cruzi: isolation and characterization of a protease. Exp. Parasit., 45: 8-18, 1978.
6. BRENER, Z. Progrés récents dans le domaine de la maladie de Chagas. Bull. Wld. Hlth. Org., 60: 845-856, 1982.
7. CALDERON, R.O. & FABRO, S.P. de Trypanosoma cruzi: fusogenic ability of membranes from cultured epimastigotes in interaction with human syncytiotrophoblast. Exp. Parasit., 56: 169-179, 1983.
8. CALDERON, R.O.; AGUERRI, A.M. & BRONIA, D.H. Trypanosoma cruzi: variable fusogenic ability by different growth phases of the epimastigote form. Exp. Parasit., 62: 453-455, 1986.
9. CAZZULO, J.J. Protein and aminoacid catabolism in Trypanosoma cruzi. Comp. Biochem. Physiol., 79B: 309-320, 1984.
10. DVORAK, B.J.A. & HYDE, T.P. Trypanosoma cruzi: interaction with vertebrate cells in vitro. I. Individual interaction at the cellular and subcellular levels. Exp. Parasit., 34: 268-283, 1973.
11. ENGEL, J.C.; DOYLE, P.S. & DVORAK, B.J.A. Trypanosoma cruzi: biological characterization of clones derived from chagasic patients. II. Quantitative analysis of the intracellular cycle. J. Protozool., 32: 80-83, 1985.
12. FABRO, S.P. de; MENDEZ, T. & IWAKAWA, L. Un método sencillo para el cultivo de órganos. Rev. Fac. Cienc. med. Univ. Córdoba, 31: 199-209, 1973.
13. FABRO, S.P. de & GOLDEMBERG, L. Modificaciones enzimáticas producidas por la enfermedad de Chagas en la embarazada y en el nińo. Medicine (B. Aires), 40 (Supl. 1): 240-241, 1980.
14. GALSKY, H.; FRIDOVICH, S.E.; WEINSTEIN, D.; DE GROOT, N.; SEGAL, S.; FOLMAN, R. & HOCHBERG, A.A. Synthesis and secretion of alkaline phosphatase in vitro from first trimester and term human placentas. Biochem J., 194: 857-866, 1981.
15. JONES, C.J.P. & FOX, H. An ultrahistochemical study of the distribution of acid and alkaline phosphatases in placentae from normal and complicated pregnancies. J. Path., 118: 143-151, 1976.
16. KARNOVSKY, M.J. A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J. Cell Biol., 27: 137A-138A, 1965.
17. LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.L. & RANDALL, R.J. Protein measurement with the phenol reagent. J. biol. Chem., 193: 265-275, 1951.
18. MARTIN, B.J.; SPICER, S.S. & SMYTHE, N.M. Cytochemical studies of the maternal surface of the syncytiotrophoblast of human early and term placenta. Anat. Rec., 178: 768-785, 1974.
19. MEIRELLES, M.N.L.; ARAÚJO JORGE, T.C. de & SOUZA, W. de Interaction of Trypanosoma cruzi with macrophages in vitro: dissociation of the attachment and internalization phases by low temperature and cytochalasin B. Z. Parasitenkd., 68: 7-14, 1982.
20. MESSER, H.H.; SHAMI, I. & COOP, D.H. Stimulation by ATP of alkaline phosphatase in placental plasma membranes. Biochim. biophys. Acta (Amst.), 391: 61-66, 1975.
21. MOYA, P.R.; VILLAGRA, L. & RISCO, J. Enfermedad de Chagas congénita: hallazgos anatomopatologicos en placenta y cordón umbilical. Rev. Fac. Cienc. méd. Univ. Córdoba, 37: 21-27, 1979.
22. RANGEL, H.A.; ARAÚJO, P.M.M.; REPKA, D. & COSTA, M.G. Trypanosoma cruzi: isolation and characterization of a proteinase. Exp. Parasit., 52: 199-209, 1981.
23. SNARY, D. Receptors and recognition mechanisms of Trypanosoma cruzi. Trans. roy. Soc. trop. Med. Hyg., 79: 587-590, 1985.
24. TAFURI, W.L.; ROCHA, A.; LOPES, E.R.; GOMES, J. & MINEO, J.R. Placentite chagásica. Apresentaçăo de um caso com estudo a microscopia óptica e eletrônica. Rev. Inst. Med. trop. S. Paulo, 26: 152-159, 1984.

Address for correspondence:

Dra. Sofía P. de Fabro

La Noria, 1262

(5000) Córdoba, Argentina

Publication Dates

Publication in this collection
11 Sept 2006
Date of issue
Dec 1990

History

Received
16 Feb 1990

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

[1] 1. ADENIYI, F.A. & OLATUNBOSUN, D.A. Origins and significance of the increased plasma alkaline phosphatase during normal pregnancy and pre-eclampsia. Brit. J. Obstet. Gynaec., 91: 857-862, 1984.

[2] 2. ANDRADE, S.G. The influence of the strain of Trypanosoma cruzi in placental infections in mice. Trans. roy. Soc. trop. Med. Hyg., 76: 123-128, 1982.

[3] 3. ANDREWS, N.W. & COLLI, W. Adhesion and interiorization of Trypanosoma cruzi in mammalian cells. J. Protozool., 29: 264-269, 1982.

[4] 4. BITTENCOURT, A.L. Congenital Chagas' disease. Amer. J. Dis. Child., 130: 97-103, 1976.

[5] 5. BONGERSZ, V. & HUNGERER, K.D. Trypanosoma cruzi: isolation and characterization of a protease. Exp. Parasit., 45: 8-18, 1978.

[6] 6. BRENER, Z. Progrés récents dans le domaine de la maladie de Chagas. Bull. Wld. Hlth. Org., 60: 845-856, 1982.

[7] 7. CALDERON, R.O. & FABRO, S.P. de Trypanosoma cruzi: fusogenic ability of membranes from cultured epimastigotes in interaction with human syncytiotrophoblast. Exp. Parasit., 56: 169-179, 1983.

[8] 8. CALDERON, R.O.; AGUERRI, A.M. & BRONIA, D.H. Trypanosoma cruzi: variable fusogenic ability by different growth phases of the epimastigote form. Exp. Parasit., 62: 453-455, 1986.

[9] 9. CAZZULO, J.J. Protein and aminoacid catabolism in Trypanosoma cruzi. Comp. Biochem. Physiol., 79B: 309-320, 1984.

[10] 10. DVORAK, B.J.A. & HYDE, T.P. Trypanosoma cruzi: interaction with vertebrate cells in vitro. I. Individual interaction at the cellular and subcellular levels. Exp. Parasit., 34: 268-283, 1973.

[11] 11. ENGEL, J.C.; DOYLE, P.S. & DVORAK, B.J.A. Trypanosoma cruzi: biological characterization of clones derived from chagasic patients. II. Quantitative analysis of the intracellular cycle. J. Protozool., 32: 80-83, 1985.

[12] 12. FABRO, S.P. de; MENDEZ, T. & IWAKAWA, L. Un método sencillo para el cultivo de órganos. Rev. Fac. Cienc. med. Univ. Córdoba, 31: 199-209, 1973.

[13] 13. FABRO, S.P. de & GOLDEMBERG, L. Modificaciones enzimáticas producidas por la enfermedad de Chagas en la embarazada y en el nińo. Medicine (B. Aires), 40 (Supl. 1): 240-241, 1980.

[14] 14. GALSKY, H.; FRIDOVICH, S.E.; WEINSTEIN, D.; DE GROOT, N.; SEGAL, S.; FOLMAN, R. & HOCHBERG, A.A. Synthesis and secretion of alkaline phosphatase in vitro from first trimester and term human placentas. Biochem J., 194: 857-866, 1981.

[15] 15. JONES, C.J.P. & FOX, H. An ultrahistochemical study of the distribution of acid and alkaline phosphatases in placentae from normal and complicated pregnancies. J. Path., 118: 143-151, 1976.

[16] 16. KARNOVSKY, M.J. A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J. Cell Biol., 27: 137A-138A, 1965.

[17] 17. LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.L. & RANDALL, R.J. Protein measurement with the phenol reagent. J. biol. Chem., 193: 265-275, 1951.

[18] 18. MARTIN, B.J.; SPICER, S.S. & SMYTHE, N.M. Cytochemical studies of the maternal surface of the syncytiotrophoblast of human early and term placenta. Anat. Rec., 178: 768-785, 1974.

[19] 19. MEIRELLES, M.N.L.; ARAÚJO JORGE, T.C. de & SOUZA, W. de Interaction of Trypanosoma cruzi with macrophages in vitro: dissociation of the attachment and internalization phases by low temperature and cytochalasin B. Z. Parasitenkd., 68: 7-14, 1982.

[20] 20. MESSER, H.H.; SHAMI, I. & COOP, D.H. Stimulation by ATP of alkaline phosphatase in placental plasma membranes. Biochim. biophys. Acta (Amst.), 391: 61-66, 1975.

[21] 21. MOYA, P.R.; VILLAGRA, L. & RISCO, J. Enfermedad de Chagas congénita: hallazgos anatomopatologicos en placenta y cordón umbilical. Rev. Fac. Cienc. méd. Univ. Córdoba, 37: 21-27, 1979.

[22] 22. RANGEL, H.A.; ARAÚJO, P.M.M.; REPKA, D. & COSTA, M.G. Trypanosoma cruzi: isolation and characterization of a proteinase. Exp. Parasit., 52: 199-209, 1981.

[23] 23. SNARY, D. Receptors and recognition mechanisms of Trypanosoma cruzi. Trans. roy. Soc. trop. Med. Hyg., 79: 587-590, 1985.

[24] 24. TAFURI, W.L.; ROCHA, A.; LOPES, E.R.; GOMES, J. & MINEO, J.R. Placentite chagásica. Apresentaçăo de um caso com estudo a microscopia óptica e eletrônica. Rev. Inst. Med. trop. S. Paulo, 26: 152-159, 1984.

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