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A Leishmania (L.) amazonensis ATP diphosphohydrolase isoform and potato apyrase share epitopes: antigenicity and correlation with disease progression

Published online by Cambridge University Press:  16 November 2007

E. S. COIMBRA
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil Departamento de Parasitologia, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
S. C. GONÇALVES-DA-COSTA
Affiliation:
Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
B. L. S. COSTA
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
N. L. L. GIAROLA
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
F. A. REZENDE-SOARES
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
M. R. FESSEL
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
A. P. FERREIRA
Affiliation:
Departamento de Parasitologia, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
C. S. F. SOUZA
Affiliation:
Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
A. L. ABREU-SILVA
Affiliation:
Departamento de Patologia, Universidade Estadual do Maranhão, São Luís, Maranhão, Brazil Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
E. G. VASCONCELOS*
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
*
*Corresponding author: Departamento de Bioquímica, ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, 36036-330, Juiz de Fora, MG, Brazil. Tel: +55 32 3229 3217. Fax: +55 32 3229 3216. E-mail: [email protected]

Summary

A Leishmania (Leishmania) amazonensis ATP diphosphohydrolase isoform was partially purified from plasma membrane of promastigotes by preparative non-denaturing polyacrylamide gel electrophoresis. SDS-PAGE followed by Western blots developed with polyclonal anti-potato apyrase antibodies identified diffuse bands of about 58–63 kDa, possibly glycosylated forms of this protein. By ELISA technique, a significantly higher total IgG antibody level against potato apyrase was found in serum from promastigote-infected mice, as compared to the uninfected mice, confirming both the existence of shared epitopes between the parasite and vegetable proteins, and the parasite ATP diphosphohydrolase antigenicity. By Western blotting, serum from amastigote-infected BALB/c mice recognizes both potato apyrase and this antigenic ATP diphosphohydrolase isoform isolated from promastigotes, suggesting that it is also expressed in the amastigote stage. The infection monitored along a 90-day period in amastigote-infected mice showed reactivity of IgG2a antibody in early steps of infection, while the disappearance of the IgG2a response and elevation of IgG1 antibody serum levels against that shared epitopes were associated with the progression of experimental leishmaniasis. This is the first observation of the antigenicity of a L. (L.) amazonensis ATP diphosphohydrolase isoform, and of the ability of cross-immunoreactivity with potato apyrase to differentiate serologically stages of leishmaniasis in infected mice.

Type
Original Articles
Copyright
Copyright © Cambridge University Press 2007

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References

REFERENCES

Abreu-Silva, A. L., Calabrese, K. S., Cupolilo, S. M. N., Cardoso, F. O., Souza, C. S. F. and Gonçalves Da Costa, S. C. (2004). Histopathological studies of visceralized Leishmania (Leishmania) amazonensis in mice experimentally infected. Veterinary Parasitology 121, 179187.CrossRefGoogle ScholarPubMed
Abreu-Silva, A. L., Calabrese, K. S., Tedesco, R. C., Mortara, R. A. and Gonçalves Da Costa, S. C. (2003). Central nervous system involvement in experimental infection with Leishmania (Leishmania) amazonensis. American Journal of Tropical Medicine and Hygiene 68, 661665.CrossRefGoogle ScholarPubMed
Almeida, R. P., Barral-Neto, M., Jesus, A. M. R., Freitas, L. A. R., Carvalho, E. M. and Barral, A. (1996). Biological behavior of Leishmania amazonensis isolated from human with cutaneous, mucosal, or visceral leishmaniasis in BALB/c mice. American Journal of Tropical Medicine and Hygiene 54, 178184.CrossRefGoogle ScholarPubMed
Asai, T., Mizuno, F., Kojima, S., Takeuchi, T., Kobayashi, A. and Suzuki, Y. (1992). High correlation in antibody titers between the Sabin-Feldman dye test and an enzyme-linked immunosorbent assay detecting immunoglobulin G antibodies to the nucleoside triphosphate hydrolase of Toxoplasma gondii. Journal of Clinical Microbiology 30, 12911293.Google Scholar
Bermudes, D., Peck, K. R., Afifi, M. A., Beckers, C. J. M. and Joiner, K. A. (1994). Tandemly repeated genes enconde nucleoside triphosphate hydrolase isoforms secreted into the parasitophorous vacuole of Toxoplasma gondii. The Journal of Biological Chemistry 269, 2925229260.CrossRefGoogle ScholarPubMed
Coimbra, E. S., Gonçalves-Da-Costa, S. C., Corte-Real, S., Freitas, F. G., Durão, A. C., Souza, C. S., Silva-Santos, M. I. and Vasconcelos, E. G. (2002). Characterization and cytochemical localization of an ATP diphosphohydrolase from Leishmania amazonensis promastigotes. Parasitology 124, 137143.Google Scholar
Do Valle, T. Z., Gaspar, E. B., Souza-Lemos, C., Souza, C. S., Marquez, F. B., Baetas-da-Cruz, W., d'Escofier, L. N., Corte-Real, S., Calabrese, K. S. and Gonçalves da Costa, S. C. (2007). Experimental Leishmania (L.) amazonensis leishmaniasis: characterization and immunogenicity of subcellular fractions. Experimental Investigations 36, 473492.Google Scholar
Faria-Pinto, P., Meirelles, M. N., Lenzi, H. L., Mota, E. M., Penido, M. L. O., Coelho, P. M. Z. and Vasconcelos, E. G. (2006). Cross-immunoreactivity between anti-potato apyrase antibodies and mammalian ATP diphosphohydrolases: potential use of the vegetal protein in experimental schistosomiasis. Memórias do Instituto Oswaldo Cruz 101, 359363.CrossRefGoogle ScholarPubMed
Faria-Pinto, P., Meirelles, M. N., Lenzi, H. L., Mota, E. M., Penido, M. L. O., Coelho, P. M. Z. and Vasconcelos, E. G. (2004). ATP diphosphohydrolase from Schistosoma mansoni egg: characterization of a new antigen. Parasitology 129, 5157.CrossRefGoogle ScholarPubMed
Gendron, F. P., Benrezzak, O., Krugh, B. W., Kong, Q., Weisman, G. A. and Beaudoin, A. R. (2002). Purine signaling and potential new therapeutic approach: possible outcomes of NTPDase inhibition. Current Drug Targets 3, 229245.Google Scholar
Gonçalves Da Costa, S. C. (2005). A modulação da resistência do hospedeiro por microorganismos. In Dinâmica das doenças infecciosas e Parasitárias (ed. Coura, J. R.), pp. 165177. Guanabara Koogan Rio de Janeiro, Brazil.Google Scholar
Gonçalves Da Costa, S. C., Barbosa-Santos, E. G. and Lagrange, P. H. (1988). Vaccination of mice against Leishmania mexicana amazonensis with microsomal fraction associated with BCG. Annales de l' Institute Pasteur 139, 143156.CrossRefGoogle ScholarPubMed
Handman, E. (2001). Leishmaniasis: current status of vaccine development. Clinical Microbiological Review 14, 229243.CrossRefGoogle ScholarPubMed
Harlow, E. and Lane, D. (1988). Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
Jones, D. E., Buxbaum, L. U. and Scott, P. (2000). IL-4-independent inhibition of IL-12 responsiveness during Leishmania amazonensis infection. Journal of Immunology 165, 364372.CrossRefGoogle ScholarPubMed
Ketllun, A. M., Urra, R., Leyton, M., Valenzuela, M. A., Mancilla, M. and Traverso-Cori, A. (1992). Purification and characterization of two isoapyrases from Solanum tuberosum var. Ultimus. Phytochemistry 31, 36913696.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophages T4. Nature, London 227, 680685.CrossRefGoogle Scholar
Lowry, O. H., Roseborough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265275.Google Scholar
Penido, M. L., Resende, D. M., Vianello, M. A., Bordin, F. H. S., Jacinto, A. A., Dias, W. D., Montesano, M. A., Nelson, D. L., Coelho, P. M. Z. and Vasconcelos, E. G. (2007). A new series of schistosomicide drugs, the alkylaminoalkanethiosulfuric acids, partially inhibit the activity of Schistosoma mansoni ATP diphosphohydrolase. European Journal of Pharmacology 570, 1017.Google Scholar
Requena, J. M., Alonso, C. and Soto, M. (2000). Evolutionarily conserved proteins as prominent immunogens during Leishmania infections. Parasitology Today 16, 246250.CrossRefGoogle ScholarPubMed
Soong, L., Chang, C.-H., Sun, J., Longley, B. J., Ruddle, N. H., Flavell, R. A. and McMahon-Pratt, D. (1997). Role of CD4+ T Cells in pathogenesis associated with Leishmania amazonensis infection. Journal of Immunology 158, 53745383.Google Scholar
Taussky, H. M. and Shorr, E. (1953). A microcolorimetric method for the determination of inorganic phosphorus. Journal of Biological Chemistry 202, 675685.CrossRefGoogle ScholarPubMed
Vasconcelos, E. G., Ferreira, S. T., Carvalho, T. M., De Souza, W., Kettlun, A. M., Mancilla, M., Valenzuela, M. A. and Verjovski-Almeida, S. (1996). Partial purification and immunohistochemical localization of an ATP diphosphohydrolase from Schistosoma mansoni. Immunological cross reactivities with potato apyrase and Toxoplasma gondii nucleoside triphosphate hydrolase. Journal of Biological Chemistry 271, 2213922145.Google Scholar
Zlotnick, G. W. and Gottlieb, M. (1986). A sensitive staining technique for the dectection of phophohydrolase activities after polyacrylamide gel electrophoresis. Analytical Biochemistry 153, 121125.CrossRefGoogle Scholar