The diagnostic importance of species specific and cross-reactive components of Taenia solium, Echinococcus granulosus, and Hymenolepis nana

Montenegro, Teresa; Gilman, Robert H.; Castillo, Rosa; Tsang, Victor; Brandt, Joy; Guevara, Angela; Sanabria, Hernan; Verastegui, Manuela; Sterling, Charles; Miranda, Elba

doi:10.1590/S0036-46651994000400005

Abstracts

Sera from patients infected with Taenia solium, Hymenolepis nana and Echinococcus granulosus were tested against homologous and heterologous parasite antigens using an ELISA assay, and a high degree of cross-reactivity was verified. To identify polypeptides responsible for this cross reactivity, the Enzyme Linked Immunoelectro Transfer Blot (EITB) was used. Sera from infected patients with T.solium, H.nana, and E.granulosus were assessed against crude, ammonium sulphate precipitated (TSASP), and lentil-lectin purified antigens of T.solium and crude antigens of.H.nana and E.granulosus. Several bands, recognized by sera from patients with T.solium, H.nana, and E.granulosus infections, were common to either two or all three cestodes. Unique reactive bands in H.nana were noted at 49 and 66 K-Da and in E.granulosus at 17-21 K-Da and at 27-32 K-Da. In the crude cysticercosis extract, a specific non glycoprotein band was present at 61-67 K-Da in addiction to specific glycoprotein bands of 50, 42, 24, 21, 18, 14, and 13 K-Da. None of the sera from patients with H.nana or E.granulosus infection cross reacted with these seven glycoprotein bands considered specific for T.solium infection.

Taenia solium; Cysticercus; antigens; Hymenolepis nana; Echinococcus granulosus; Hydatid Cysts; SDS-PAGE; Electrophoresis; Immunoblotting

Soros de pacientes infectados com Taenia solium, Hymenolepis nana e Echinococcus granulosus foram testados contra antígenos parasitários homólogos e heterólogos usando o teste de ELISA e foi verificado alto grau de reatividade cruzada. Para identificar os polipetídeos responsáveis por esta reatividade cruzada foi utilizado o teste "Enzyme Linked Immunoelectro Transfer Blot (EITB)". Soros de pacientes infectados por T.solium, H.nana, e E.granulosus foram colocados em contato com precipitado de sulfato de amônia e antígenos não purificados de T.solium e os de H.nana e E.granulosus. Várias bandas reconhecidas pelos soros de pacientes com infecção por T.solium, H.nana e E.granulosus foram comuns a dois ou três destes cestódeos. Uma única banda foi notada em H.nana a 49 e 66K-Da e no E.granulosus a 17-21 K-Da e 27-32 K-Da. No extrato não purificado de cisticercose uma banda específica não glicoproteica estava presente a 61-67 K-Da além das bandas de glicoproteínas específicas de 50, 42, 24, 21, 18, 14 e 13 K-Da. Nenhum destes soros de pacientes com infecção por H.nana ou E.granulosus reagiu de forma cruzada com estas sete bandas de glicoproteína consideradas específicas à infecção por T.solium

SEROLOGY

The diagnostic importance of species specific and cross-reactive components of Taenia solium, Echinococcus granulosus, and Hymenolepis nana

Importância diagnóstica da reação cruzada espécie-específica de componentes da Taenia solium, Echinococcus granulosus e Hymenolepis nana

Teresa Montenegro^I; Robert H. Gilman^II; Rosa Castillo^I; Victor Tsang^III; Joy Brandt^III; Angela Guevara^I; Hernan Sanabria^I; Manuela Verastegui^I; Charles Sterling^IV; Elba Miranda^I

^IThe Universidad Peruana Cayetano Heredia

^IIThe Johns Hopkins University

^IIIThe Centers for Disease Control

^IVThe University of Arizona

^{Address for Correspondence} Address for Correspondence: MSc. Teresa Montenegro Depto. Microbiologia, Universidad Peruana Cayetano Heredia Av. Honorio Delgado nº430 Urb. Ingenieria, San Martin de Porras Casilla Postal 4314, Lima-Peru

SUMMARY

Sera from patients infected with Taenia solium, Hymenolepis nana and Echinococcus granulosus were tested against homologous and heterologous parasite antigens using an ELISA assay, and a high degree of cross-reactivity was verified. To identify polypeptides responsible for this cross reactivity, the Enzyme Linked Immunoelectro Transfer Blot (EITB) was used.

Sera from infected patients with T.solium, H.nana, and E.granulosus were assessed against crude, ammonium sulphate precipitated (TSASP), and lentil-lectin purified antigens of T.solium and crude antigens of.H.nana and E.granulosus.

Several bands, recognized by sera from patients with T.solium, H.nana, and E.granulosus infections, were common to either two or all three cestodes. Unique reactive bands in H.nana were noted at 49 and 66 K-Da and in E.granulosus at 17-21 K-Da and at 27-32 K-Da. In the crude cysticercosis extract, a specific non glycoprotein band was present at 6167 K-Da in addiction to specific glycoprotein bands of 50, 42, 24, 21, 18, 14, and 13 K-Da.

None of the sera from patients with H.nana or E.granulosus infection cross reacted with these seven glycoprotein bands considered specific for T.solium infection.

Keywords:Taenia solium; Cysticercus; antigens; Hymenolepis nana; Echinococcus granulosus; Hydatid Cysts; SDS-PAGE; Electrophoresis; Immunoblotting.

RESUMO

Soros de pacientes infectados com Taenia solium, Hymenolepis nana e Echinococcus granulosus foram testados contra antígenos parasitários homólogos e heterólogos usando o teste de ELISA e foi verificado alto grau de reatividade cruzada. Para identificar os polipetídeos responsáveis por esta reatividade cruzada foi utilizado o teste "Enzyme Linked Immunoelectro Transfer Blot (EITB)".

Soros de pacientes infectados por T.solium, H.nana, e E.granulosus foram colocados em contato com precipitado de sulfato de amônia e antígenos não purificados de T.solium e os de H.nana e E.granulosus.

Várias bandas reconhecidas pelos soros de pacientes com infecção por T.solium, H.nana e E.granulosus foram comuns a dois ou três destes cestódeos. Uma única banda foi notada em H.nana a 49 e 66K-Da e no E.granulosus a 17-21 K-Da e 27-32 K-Da. No extrato não purificado de cisticercose uma banda específica não glicoproteica estava presente a 61-67 K-Da além das bandas de glicoproteínas específicas de 50, 42, 24, 21, 18, 14 e 13 K-Da. Nenhum destes soros de pacientes com infecção por H.nana ou E.granulosus reagiu de forma cruzada com estas sete bandas de glicoproteína consideradas específicas à infecção por T.solium.

Full text available only in PDF format.

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ACKNOWLEDGMENTS

This work was supported by the Consejo Nacional de Ciencia y Tecnologia (CONCYTEC), Peru., The International Development Research Center (IDRC), Canada., and the PSTC No.7284 Grants.

Recebido para publicação em 31/08/1993

Aceito para publicação em 04/05/1994.

1. BERNTZEN, A. & KOGE, K. - In vitro hatching of oncospheres of four hymenolepidid cestodes. J. Parasit., 51: 235-242, 1965.
2. BRANDFORD, M. - A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye-binding. Ann. Biochem., 72: 248-254, 1976.
3. CASTILLO, R.; GRADOS, P.; CARCAMO, C. et. al. - Effect of treatment on serum antibody to Hymenolepis nana detected by Enzyme-Linked Immunosorbent Assay. J. clin. Microbiol., 29: 413-414, 1991.
4. CODER-VANN, M.; BROWN, P. & GAJDUSEK, C. - Serodiagnosis of human cysticercosis using a chromatofocused antigenic preparation of Taenia solium cysticerci in an enzyme-linked immunosorbent assay (ELISA). Trans. roy. Soc. trop. Med. Hyg., 78: 492-496, 1984.
5. DIAZ, F.; GARCIA, H.; GILMAN, R. et. al. - Epidemiology of Taeniasis and Cysticercosis in a Peruvian village. Amer. J. Epidem., 135: 875-882, 1992.
6. DIWAN, A.; COKER-VANN, M. & BROWN, M. - Enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to cysticerci of Taenia solium. Amer. J. trop. Med. Hyg., 31: 364-369, 1982.
7. ESPINOZA, B.; FLISSER, A.; PLANCARTE, A. & LARRALDE, C. - Immunodiagnosis of human cysticercosis: ELISA and immunoelectrophoresis. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 163-170.
8. ESPINOZA, B.; RUIZ-PALACIOS, G. & TOVAR, A. - Characterization by enzyme-linked immunosorbent assay of the humoral immune response in patients with neurocysticercosis and its application in immunodiagnosis. J.clin. Microbiol., 24: 536-541, 1986.
9. ESTRADA, J.J. & KUHN, R.E. - Immunochemical detection of antigens of larval Taenia solium and anti-larval antibodies in the cerebrospinal fluid of patients with neurocysticercosis. J. Neurol. Sci., 71: 39-48, 1985.
10. FLISSER, A. & LARRALDE, C. - Cysticercosis. In: WALLS, K., ed. Immunodiagnosis of parasitic diseases, helminthic diseases. New York, Academic Press, 1986. v.1, p. 109-161.
11. GREEN, A. & HUGHES, W. - Protein fractionation on the basis solubility in aqueous solutions of salts and organic solvents. In: COLOWICK, S., ed. - Methods in enzymology. New York, Academic Press, 1955. p. 67-78.
12. GROGL, M.; ESTRADA, J.; MACDONALD, G. & KUHN, R. - Antigen-antibody analyses in neurocysticercosis. J. Parasit., 71: 433-442, 1985.
13. HARRIS, W.G. - Antibody response to tapeworm (Hymenolepis diminuta) in the rat. Nature, 246: 21, 1973.
14. LARRALDE, C.; LACLETTE, J.P. & OWEN, S. - Reliable serology of Taenia solium cysticercosis with antigens from cyst vesicular fluid: ELISA and hemagglutination tests. Amer. J. trop. Med.Hyg., 35: 965-973, 1986.
15. MOHAMMAD, I.; HEINER, D.; MILLER, B.; GOLDBERG, M. & KAGAN, I. - Enzyme-linked immunosorbent assay for the diagnosis of cerebral cysticercosis. J. clin. Microbiol., 20: 775-779, 1984.
16. MAHALANABIS, D.; SIMPSON, T. & CHAKRABORTY, M. - Malabsorption of water miscible vitamin A in children with giardiasis and ascariasis. Amer. J. clin. Nutr., 32: 313-318, 1979.
17. MAHAJAN, R. - Geographical distribution of human cysticercosis. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 39-46.
18. NIETO, D. - Cysticercosis of the nervous system. Diagnosis by means of the spinal fluid complement fixation test. Neurology, 6: 725-738, 1956.
19. PAMMENTER, M. & RUSSOUW, E. - The value of an antigenic fraction of Cysticercus cellulosae in the serodiagnosis of cysticercosis. Ann. trop. Med. Parasit., 81: 117-123, 1987.
20. SCHENONE, H.; VILLAROEL, F.; ROJAS, A. & RAMIREZ, R. - Epidemiology of human cysticercosis in Latin America. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 25-38.
21. SHEPHERED, J. & MANUS, D. - Specific and cross-reactive antigen of Echinococcus granulosus hydatid cyst fluid. Molec. Biochem. Parasit., 25: 143-154, 1987.
22. TSANG, V.; HANCOCK, K.; WILSON, M. et. al. - Immunoelectrotransfer Blot Technique (Western Blot) for Human T-Lymphotropic Virus Type III/Lymphadenopathy-Associated Virus (HTLV-III/LAV) Antibodies. Atlanta, Centers for Disease Control, 1986. p. 125. (Monograph: Immunology Series nş 15).
23. TSANG, V.; BRAND, J. & BOYER, A. - An enzyme-linked immunoelectrotransfer blot assay and glycoprotein antigens for diagnosing human cysticercosis (Taenia solium). J. infect. Dis., 159: 50-59, 1989.
24. VALERA-DÍAZ, V.; NAQUIRA, F.; COLTORTI, E. et. al. - La hidatidosis en el Peru: importancia del inmunodiagnostico y de los estudios seroepidemiologicos. Acta med. peru., 11: 21-28, 1984.
25. VALERA-DÍAZ, V. & COLTORTI, E. - Técnicas para el diagnósitico inmunológico de la hidatidosis humana. Buenos Aires, Centro Panamericano de Zoonosis, OPS/OMS, 1974. p. 11-13.(Monografias Científicas y Técnicas, nş 7).
26. ZENTENO, G. - A classification of human cysticercosis. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 107-126.

Address for Correspondence:

MSc. Teresa Montenegro

Depto. Microbiologia, Universidad Peruana Cayetano Heredia

Av. Honorio Delgado nº430

Urb. Ingenieria, San Martin de Porras

Casilla Postal 4314, Lima-Peru

Publication Dates

Publication in this collection
20 Sept 2006
Date of issue
Aug 1994

History

Accepted
04 May 1994
Received
31 Aug 1993

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

[1] 1. BERNTZEN, A. & KOGE, K. - In vitro hatching of oncospheres of four hymenolepidid cestodes. J. Parasit., 51: 235-242, 1965.

[2] 2. BRANDFORD, M. - A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye-binding. Ann. Biochem., 72: 248-254, 1976.

[3] 3. CASTILLO, R.; GRADOS, P.; CARCAMO, C. et. al. - Effect of treatment on serum antibody to Hymenolepis nana detected by Enzyme-Linked Immunosorbent Assay. J. clin. Microbiol., 29: 413-414, 1991.

[4] 4. CODER-VANN, M.; BROWN, P. & GAJDUSEK, C. - Serodiagnosis of human cysticercosis using a chromatofocused antigenic preparation of Taenia solium cysticerci in an enzyme-linked immunosorbent assay (ELISA). Trans. roy. Soc. trop. Med. Hyg., 78: 492-496, 1984.

[5] 5. DIAZ, F.; GARCIA, H.; GILMAN, R. et. al. - Epidemiology of Taeniasis and Cysticercosis in a Peruvian village. Amer. J. Epidem., 135: 875-882, 1992.

[6] 6. DIWAN, A.; COKER-VANN, M. & BROWN, M. - Enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to cysticerci of Taenia solium. Amer. J. trop. Med. Hyg., 31: 364-369, 1982.

[7] 7. ESPINOZA, B.; FLISSER, A.; PLANCARTE, A. & LARRALDE, C. - Immunodiagnosis of human cysticercosis: ELISA and immunoelectrophoresis. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 163-170.

[8] 8. ESPINOZA, B.; RUIZ-PALACIOS, G. & TOVAR, A. - Characterization by enzyme-linked immunosorbent assay of the humoral immune response in patients with neurocysticercosis and its application in immunodiagnosis. J.clin. Microbiol., 24: 536-541, 1986.

[9] 9. ESTRADA, J.J. & KUHN, R.E. - Immunochemical detection of antigens of larval Taenia solium and anti-larval antibodies in the cerebrospinal fluid of patients with neurocysticercosis. J. Neurol. Sci., 71: 39-48, 1985.

[10] 10. FLISSER, A. & LARRALDE, C. - Cysticercosis. In: WALLS, K., ed. Immunodiagnosis of parasitic diseases, helminthic diseases. New York, Academic Press, 1986. v.1, p. 109-161.

[11] 11. GREEN, A. & HUGHES, W. - Protein fractionation on the basis solubility in aqueous solutions of salts and organic solvents. In: COLOWICK, S., ed. - Methods in enzymology. New York, Academic Press, 1955. p. 67-78.

[12] 12. GROGL, M.; ESTRADA, J.; MACDONALD, G. & KUHN, R. - Antigen-antibody analyses in neurocysticercosis. J. Parasit., 71: 433-442, 1985.

[13] 13. HARRIS, W.G. - Antibody response to tapeworm (Hymenolepis diminuta) in the rat. Nature, 246: 21, 1973.

[14] 14. LARRALDE, C.; LACLETTE, J.P. & OWEN, S. - Reliable serology of Taenia solium cysticercosis with antigens from cyst vesicular fluid: ELISA and hemagglutination tests. Amer. J. trop. Med.Hyg., 35: 965-973, 1986.

[15] 15. MOHAMMAD, I.; HEINER, D.; MILLER, B.; GOLDBERG, M. & KAGAN, I. - Enzyme-linked immunosorbent assay for the diagnosis of cerebral cysticercosis. J. clin. Microbiol., 20: 775-779, 1984.

[16] 16. MAHALANABIS, D.; SIMPSON, T. & CHAKRABORTY, M. - Malabsorption of water miscible vitamin A in children with giardiasis and ascariasis. Amer. J. clin. Nutr., 32: 313-318, 1979.

[17] 17. MAHAJAN, R. - Geographical distribution of human cysticercosis. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 39-46.

[18] 18. NIETO, D. - Cysticercosis of the nervous system. Diagnosis by means of the spinal fluid complement fixation test. Neurology, 6: 725-738, 1956.

[19] 19. PAMMENTER, M. & RUSSOUW, E. - The value of an antigenic fraction of Cysticercus cellulosae in the serodiagnosis of cysticercosis. Ann. trop. Med. Parasit., 81: 117-123, 1987.

[20] 20. SCHENONE, H.; VILLAROEL, F.; ROJAS, A. & RAMIREZ, R. - Epidemiology of human cysticercosis in Latin America. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 25-38.

[21] 21. SHEPHERED, J. & MANUS, D. - Specific and cross-reactive antigen of Echinococcus granulosus hydatid cyst fluid. Molec. Biochem. Parasit., 25: 143-154, 1987.

[22] 22. TSANG, V.; HANCOCK, K.; WILSON, M. et. al. - Immunoelectrotransfer Blot Technique (Western Blot) for Human T-Lymphotropic Virus Type III/Lymphadenopathy-Associated Virus (HTLV-III/LAV) Antibodies. Atlanta, Centers for Disease Control, 1986. p. 125. (Monograph: Immunology Series nş 15).

[23] 23. TSANG, V.; BRAND, J. & BOYER, A. - An enzyme-linked immunoelectrotransfer blot assay and glycoprotein antigens for diagnosing human cysticercosis (Taenia solium). J. infect. Dis., 159: 50-59, 1989.

[24] 24. VALERA-DÍAZ, V.; NAQUIRA, F.; COLTORTI, E. et. al. - La hidatidosis en el Peru: importancia del inmunodiagnostico y de los estudios seroepidemiologicos. Acta med. peru., 11: 21-28, 1984.

[25] 25. VALERA-DÍAZ, V. & COLTORTI, E. - Técnicas para el diagnósitico inmunológico de la hidatidosis humana. Buenos Aires, Centro Panamericano de Zoonosis, OPS/OMS, 1974. p. 11-13.(Monografias Científicas y Técnicas, nş 7).

[26] 26. ZENTENO, G. - A classification of human cysticercosis. In: FLISSER, A., ed. Cysticercosis. Present state of knowledge and perspectives. New York, Academic Press, 1982. p. 107-126.

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