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ATP diphosphohydrolase from Schistosoma mansoni egg: characterization and immunocytochemical localization of a new antigen

Published online by Cambridge University Press:  10 June 2004

P. FARIA-PINTO ,
M. N. L. MEIRELLES ,
H. L. LENZI ,
E. M. MOTA ,
M. L. O. PENIDO ,
P. M. Z. COELHO  and
E. G. VASCONCELOS
P. FARIA-PINTO
Affiliation:
Departamento de Bioquímica/ICB, Universidade Federal de Juiz de Fora, 36015-400 Juiz de Fora, MG, Brasil Departamento de Ultraestrutura e Biologia Celular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brasil
M. N. L. MEIRELLES
Affiliation:
Departamento de Ultraestrutura e Biologia Celular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brasil
H. L. LENZI
Affiliation:
Departamento de Patologia, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brasil
E. M. MOTA
Affiliation:
Departamento de Patologia, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brasil
M. L. O. PENIDO
Affiliation:
Departamento de Bioquímica/ICB, Universidade Federal de Juiz de Fora, 36015-400 Juiz de Fora, MG, Brasil
P. M. Z. COELHO
Affiliation:
Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, FIOCRUZ, and Sta Casa de Misericórdia de Belo Horizonte, Belo Horizonte, MG, Brasil
E. G. VASCONCELOS
Affiliation:
Departamento de Bioquímica/ICB, Universidade Federal de Juiz de Fora, 36015-400 Juiz de Fora, MG, Brasil
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Abstract

The fact that the Schistosoma mansoni egg has two ATP diphosphohydrolase (EC 3.6.1.5) isoforms with different net charges and an identical molecular weight of 63000, identified by non-denaturing polyacrylamide gel electrophoresis and immunological cross-reactivity with potato apyrase antibodies, is shown. In soluble egg antigen (SEA), only the isoform with the lower net negative charge was detected and seemed to be the predominant species in this preparation. By confocal fluorescence microscopy, using anti-potato apyrase antibodies, the S. mansoni egg ATP diphosphohydrolase was detected on the external surface of miracidium and in von Lichtenberg's envelope. Intense fluorescence was also seen in the outer side of the egg-shell, entrapped by the surface microspines, suggesting that a soluble isoform is secreted. ATP diphosphohydrolase antigenicity was tested using the vegetable protein as antigen. The purified potato apyrase was recognized in Western blots by antibodies present in sera from experimentally S. mansoni-infected mice. In addition, high levels of IgG anti-ATP diphosphohydrolase antibodies were detected by ELISA in the same sera. This work represents the first demonstration of antigenic properties of S. mansoni ATP diphosphohydrolase and immunological cross-reactivity between potato apyrase and sera from infected individuals.

Keywords

ATP diphosphohydrolasepotato apyraseSchistosoma mansoni eggimmunological cross-reactivityantigenicityimmunolocalizationimmunocytochemistry
Type
Research Article
Information
Parasitology , Volume 129 , Issue 1 , July 2004 , pp. 51 - 57
Copyright
© 2004 Cambridge University Press

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References

REFERENCES

ASHTON, P. D., HARROP, R., SHAH, B. & WILSON, R. A. ( 2001). The schistosome egg: development and secretions. Parasitology 122, 329–338.CrossRefGoogle Scholar
BOROS, D. L. & WARREN, K. S. ( 1970). Delayed hypersensitivity-type granuloma formation and dermal reaction induced and elicited by a soluble factor isolated from Schistosoma mansoni eggs. Journal of Experimental Medicine 132, 488–507.CrossRefGoogle Scholar
CESARI, I. M., BALLEN, D. E., PERRONE, T., ORIOL, O., HOEBEKE, J. & BOUT, D. ( 2000). Enzyme activities in Schistosoma mansoni soluble egg antigen. Journal of Parasitology 86, 1137–1140.CrossRefGoogle Scholar
COIMBRA, E. S., GONÇALVES DA COSTA, S. C., CORTE-REAL, S., FREITAS, F. G. R., DURÃO, A., SOUZA, C. S. F., SILVA SANTOS, M. I. & VASCONCELOS, E. G. ( 2002). Characterization and cytochemical localization of an ATP diphosphohydrolase from Leishmania amazonensis promastigotes. Parasitology 124, 137–143.CrossRefGoogle Scholar
DeMARCO, R., KOWALTOWSKI, A. T., MORTARA, R. A. & VERJOVSKI-ALMEIDA, S. ( 2003). Molecular and characterization of Schistosoma mansoni ATP diphosphohydrolase. Biochemical and Biophysical Research Communications 307, 831–838.CrossRefGoogle Scholar
FILIPPINI, A., TAFFS, R. E., AGUI, T. & SITKOVSKY, M. V. ( 1990). Ecto-ATPase activity in cytolytic T-lymphocytes. Journal of Biological Chemistry 265, 334–340.Google Scholar
GENDRON, F. P., BENREZZAK, O., KRUGH, B. W., KONG, Q., WEISMAN, G. A. & BEAUDOIN, A. R. ( 2002). Purine signaling and potential new therapeutic approach: possible outcomes of NTPDase inhibition. Current Drug Targets 3, 229–245.CrossRefGoogle Scholar
HANDA, M. & GUIDOTTI, G. ( 1996). Purification and cloning of a soluble ATP diphosphohydrolase (apyrase) from potato tubers (Solanum tuberosum). Biochemical and Biophysical Research Communications 218, 916–923.CrossRefGoogle Scholar
HARLOW, E. & LANE, D. ( 1988). Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
KETLLUN, A. M., URRA, R., LEYTON, M., VALENZUELA, M. A., MANCILLA, M. & TRAVERSO-CORI, A. ( 1992). Purification and characterization of two isoapyrases from Solanum tuberosum var. Ultimus. Phytochemistry 31, 3691–3696.CrossRefGoogle Scholar
LAEMMLI, U. K. ( 1970). Cleavage of structural proteins during the assembly of the head of bacteriophages T4. Nature, London 227, 680–685.CrossRefGoogle Scholar
LOWRY, O. H., ROSEBROUGH, N. J., FARR, A. L. & RANDALL, R. J. ( 1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265–275.Google Scholar
SMITHERS, S. R. & TERRY, R. J. ( 1965). The infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of adult worms. Parasitology 55, 695–700.CrossRefGoogle Scholar
STADECKER, M. J., HERNANDEZ, H. J. & ASAHI, H. ( 2001). The identification and characterization of new immunogenic egg components: implications for evaluation and control of the immunopathogenic T cell response in schistosomiasis. Memórias do Instituto Oswaldo Cruz 96, 29–33.CrossRefGoogle Scholar
TAUSSKY, H. M. & SHORR, E. ( 1953). A microcolorimetric method for the determination of inorganic phosphorus. Journal of Biological Chemistry 202, 675–685.Google Scholar
VARGAFTIG, B. B., CHIGNARD, M. & BENEVISTE, J. ( 1981). Present concepts on the mechanism of platelet aggregation. Biochemical Pharmacology 30, 263–271.CrossRefGoogle Scholar
VASCONCELOS, E. G., FERREIRA, S. T., CARVALHO, T. M. U., SOUZA, W., KETTLUN, A. M., MANCILLA, M., VALENZUELA, M. A. & VERJOVSKI-ALMEIDA, S. ( 1996). Partial purification and immunohistochemical localization of ATP diphosphohydrolase from Schistosoma mansoni. Journal of Biological Chemistry 271, 22139–22145.CrossRefGoogle Scholar