Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (Viannia) braziliensis

Vexenat, Ana de Cássia; Santana, Jaime M.; Teixeira, Antonio R.L.

doi:10.1590/S0036-46651996000300003

Abstracts

We have detected antibodies, in the sera of Chagas disease, Kala-azar and Mucocutaneous leishmaniasis patients, that bind multiple antigens shared between the three causative agents. The Chagas disease sera showed 98 to 100% positive results by ELISA when the Leishmania braziliensis and Leishmania chagasi antigens were used, respectively. The Kala-azar sera showed 100% positive results with Trypanosoma cruzi or L. braziliensis antigens by immunofluorescence assays. The antibodies in the sera of Mucocutaneous leishmaniasis patients showed 100% positive results by ELISA assays with T. cruzi or L. chagasi antigens. Furthermore, the direct agglutination of L. chagasi promastigotes showed that 95% of Kala-azar and 35% of Mucocutaneous leishmaniasis sera agglutinated the parasite in dilutions above 1:512. In contrast, 15% of Chagas sera agglutinated the parasite in dilutions 1:16 and below. Western blot analysis showed that the Chagas sera that formed at least 24 bands with the T. cruzi also formed 13 bands with the L. chagasi and 17 bands with the L. braziliensis. The Kala-azar sera that recognized at least 29 bands with the homologous antigen also formed 14 bands with the T. cruzi and 10 bands with the L. braziliensis antigens. Finally, the Mucocutaneous leishmaniasis sera that formed at least 17 bands with the homologous antigen also formed 10 bands with the T. cruzi and four bands with the L. chagasi antigens. These results indicate the presence of common antigenic determinants in several protozoal proteins and, therefore, explain the serologic cross-reactions reported here.

Kinetoplastid protozoa; Trypanosoma cruzi; Leishmania chagasi; Leishmania (Viannia) braziliensis; Cross-reactive antibodies; Human infections

Foram detectados anticorpos, nos soros de pacientes com doença de Chagas, Calazar e Leishnnaniose cutâneo-mucosa, que se ligam a antígenos compartilhados pelos três agentes causais. Os pacientes chagásicos mostraram 98 a 100% de soropositividade pelo ELISA quando antígenos de Leishmania braziliensis e de Leishmania chagasi foram usados, respectivamente. Os soros de Calazar mostraram resultados positivos com antígenos de Trypanosoma cruzi ou com L. braziliensis pela imunofluorescência. Ademais, os anticorpos nos soros de pacientes com Leishmaniose cutâneo-mucosa tinham 100% de resultados positivos pelo ELISA com antígenos de T. cruzi ou de L. chagasi. Ademais, a aglutinação direta de promastigotas de L. chagasi mostrou que 95% dos soros de Calazar e 35% de Leishmaniose cutâneo-mucosa algutinaram o parasito em diluições acima de 1:512. Em contraste, 15% dos soros chagásicos aglutinaram o parasito na diluição de 1:16 ou abaixo. Análises pelo Western blot mostraram que os soros chagásicos que formaram pelo menos 24 bandas com antígeno de T. cruzi também foramaram 13 bandas com antígeno de L. chagasi e 17 bandas com L. braziliensis. Os soros de Calazar que reconheceram pelo menos 29 bandas com o antígeno homólogo também formaram 14 bandas com o T. cruzi e 10 bandas com L. braziliensis. Finalmente, os soros de Leishmaniose cutâneo-mucosa que formaram pelo menos 17 bandas com o antígeno homólogo também formaram 10 bandas com os antígenos de T. cruzi e quatro com L. braziliensis. Esses resultados indicam a presença de determinantes antigênicos comuns em várias proteínas desses protozoários e, portanto, explicam as reações sorológicas cruzadas descritas aqui.

IMMUNOLOGY

Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (Viannia) braziliensis

Reatividade cruzada de anticorpos em pacientes com infecções pelos protozoários Trypanosoma cruzi, Leishmania chagasi e Leishmania (Viannia) braziliensis

Ana de Cássia Vexenat; Jaime M. Santana; Antonio R.L. Teixeira

Laboratório Multidisciplinar de Pesquisa em Doença de Chagas. Faculdade de Ciências da Saúde da Universidade de Brasília. Caixa Postal 04685, 70919-970 Brasilia, Brazil

^{Correspondence to} Correspondence to: Dr. Antonio R.L. Teixeira Laboratório Multidisciplinar de Pesquisa em Doença de Chagas, Faculdade de Ciências da Saúde, Universidade de Brasília Caixa Postal 04685 70919-970 Brasília, DF, Brasil FAX: (061) 273-4645

SUMMARY

We have detected antibodies, in the sera of Chagas disease, Kala-azar and Mucocutaneous leishmaniasis patients, that bind multiple antigens shared between the three causative agents. The Chagas disease sera showed 98 to 100% positive results by ELISA when the Leishmania braziliensis and Leishmania chagasi antigens were used, respectively. The Kala-azar sera showed 100% positive results with Trypanosoma cruzi or L. braziliensis antigens by immunofluorescence assays. The antibodies in the sera of Mucocutaneous leishmaniasis patients showed 100% positive results by ELISA assays with T. cruzi or L. chagasi antigens. Furthermore, the direct agglutination of L. chagasi promastigotes showed that 95% of Kala-azar and 35% of Mucocutaneous leishmaniasis sera agglutinated the parasite in dilutions above 1:512. In contrast, 15% of Chagas sera agglutinated the parasite in dilutions 1:16 and below.

Western blot analysis showed that the Chagas sera that formed at least 24 bands with the T. cruzi also formed 13 bands with the L. chagasi and 17 bands with the L. braziliensis. The Kala-azar sera that recognized at least 29 bands with the homologous antigen also formed 14 bands with the T. cruzi and 10 bands with the L. braziliensis antigens. Finally, the Mucocutaneous leishmaniasis sera that formed at least 17 bands with the homologous antigen also formed 10 bands with the T. cruzi and four bands with the L. chagasi antigens. These results indicate the presence of common antigenic determinants in several protozoal proteins and, therefore, explain the serologic cross-reactions reported here.

Keywords: Kinetoplastid protozoa; Trypanosoma cruzi; Leishmania chagasi; Leishmania (Viannia) braziliensis; Cross-reactive antibodies; Human infections.

RESUMO

Foram detectados anticorpos, nos soros de pacientes com doença de Chagas, Calazar e Leishnnaniose cutâneo-mucosa, que se ligam a antígenos compartilhados pelos três agentes causais. Os pacientes chagásicos mostraram 98 a 100% de soropositividade pelo ELISA quando antígenos de Leishmania braziliensis e de Leishmania chagasi foram usados, respectivamente. Os soros de Calazar mostraram resultados positivos com antígenos de Trypanosoma cruzi ou com L. braziliensis pela imunofluorescência. Ademais, os anticorpos nos soros de pacientes com Leishmaniose cutâneo-mucosa tinham 100% de resultados positivos pelo ELISA com antígenos de T. cruzi ou de L. chagasi. Ademais, a aglutinação direta de promastigotas de L. chagasi mostrou que 95% dos soros de Calazar e 35% de Leishmaniose cutâneo-mucosa algutinaram o parasito em diluições acima de 1:512. Em contraste, 15% dos soros chagásicos aglutinaram o parasito na diluição de 1:16 ou abaixo.

Análises pelo Western blot mostraram que os soros chagásicos que formaram pelo menos 24 bandas com antígeno de T. cruzi também foramaram 13 bandas com antígeno de L. chagasi e 17 bandas com L. braziliensis. Os soros de Calazar que reconheceram pelo menos 29 bandas com o antígeno homólogo também formaram 14 bandas com o T. cruzi e 10 bandas com L. braziliensis. Finalmente, os soros de Leishmaniose cutâneo-mucosa que formaram pelo menos 17 bandas com o antígeno homólogo também formaram 10 bandas com os antígenos de T. cruzi e quatro com L. braziliensis. Esses resultados indicam a presença de determinantes antigênicos comuns em várias proteínas desses protozoários e, portanto, explicam as reações sorológicas cruzadas descritas aqui.

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19. TEIXEIRA, A.R.L. - The stercorarian Trypanosomes. In: SOULSBY, E.J.L., ed. Immune response in parasitic infections: immunology, immunopathology and immunoprophylaxis. Boca Raton, CRC Press, 1987. v. 3, p. 25-118.

Recebido para publicação em 30/10/1995

Aceito para publicação em 07/05/1996

Grant support: FINEP, Financiadora de Estudos e Projetos.

1. ALENCAR, J.E.; ALMEIDA, Y.M.; SILVA, Z.F.; PAIVA, S.A. & FONSECA, M.F. - Aspectos atuais do Calazar no Ceará. Rev. bras. Malar., 25: 27-53, 1974.
2. CAMARGO, M.E. - Fluorescent antibody test for the serodiagnosis of American Trypanosomiasis: technical modification employing preserved culture forms of Trypanosoma cruzi in a slide test. Rev. Inst. Med. trop. S. Paulo, 8: 227-234, 1966.
3. CAMARGO, M.E. & REBONATO, C. - Cross-reactivity in immunofluorescence test for Trypanosoma and Leishmania antibodies. A simple inhibition procedure to ensure specific results. Amer J. trop. Med. Hyg., 18: 500-505, 1969.
4. CHILLER, T.M.; SAMUDIO, M.A. & ZOULEK, G. - IgG antibody reactivity with Trypanosoma cruzi and Leishmania antigens in sera of patients with Chagas' disease and leishmaniasis. Amer. J. trop. Med. Hyg., 43: 249-259, 1990.
5. CUBA-CUBA, C.A.; MARSDEN, P.D.; BARRETO, A.C. et al. - Parasitologic and immunologic diagnosis of American (mucocutaneous) leishmaniasis. Bull. Pan Amer. Hlth. Org., 15: 249-259, 1981.
6. EVANS, D.A. Leishmania reference strains. Parasit. today, 1: 172-173, 1985.
7. GRIMALDI Jr., G.; TESHL, R.B. & MCMAHON-PRATT, D. - A review of the geographic distribution and epidemiology of leishmaniasis in the New World. Amer. J. trop. Med. Hyg., 41: 687-725, 1989.
8. HARITH, A.E.; KOLK, A.H.J.; MINTER GOEDBLOED, E.; KAGER, P.A. & KOLK, A.H.J. - Improvement of a direct agglutination test for field studies of visceral leishmaniasis. J. clin. Microbiol., 26: 1321-1325, 1988.
9. LAEMMLI, U.K. - Cleavage of structural proteins during assembly of the head of bacteriophage. Nature, 227: 680-685, 1970.
10. LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.L. & RANDALL, R.J. - Protein measurement with Folin phenol reagent. J. biol. Chem., 193: 265-275, 1951.
11. MALCHIODI, E.L.; CHIARAMONTE, M.G.; TARANTO, N.J.; ZWIRNER, N.W. & MARGNI, R.A. - Cross-reactivity studies and differential diagnosis of human infections caused by Trypanosoma cruzi and Leishmania spp; use of immunobloting and ELISA with a purified antigen (Ag163Bb). Clin. exp. Immunol., 97: 417-423, 1994.
12. MARSDEN, P.D. - Leishmaniasis. New Engl. J. Med., 300: 350-352, 1974.
13. MASUDA, A.; NASCIMENTO, S.F.; GUERRA, C.S.; PARANHOS, G.S. & FERREIRA, A.W. - Analysis of the specificity of human antibodies to antigens of Leishmania braziliensis braziliensis. Rev. Inst. Med. trop. S. Paulo, 31: 228-234, 1989.
14. MOTT, K.E.; LEHMAN Jr., J.S.; HOFF, R. et al. - The epidemiology and household distribution of seroreactivity to Trypanosoma cruzi in a rural community in northeast Brazil. Amer J. trop. Med. Hyg., 25: 552-562, 1976.
15. NEVA, F.A. & GAM, A. - A complement-fixing antigen from Trypanosoma cruzi grown in cell cultures. Amer. J. trop. Med. Hyg., 26: 37-46, 1977.
16. SHAW, J.J. & LAINSON, R. - Ecology and epidemiology: new world. In: PETERS, W. & KILLICK-KENDRICK, R., ed. The leishmaniasis in biology and medicine. London, Academic-Press, 1987. v. 1, p. 291-363.
17. TEIXEIRA, A.R.L. & PEREIRA, L.M. - Discrepâncias entre resultados de tręs reaçőes sorológicas empregadas para diagnóstico da Doença de Chagas. Rev. bras. Biol., 42: 789-795, 1981.
18. TEIXEIRA, A.R.L.; JUNQUEIRA Jr., L.F.; SOLÓRZANO, E. & ZAPPALA, M. - Doença de Chagas experimental em coelhos isogęnicos III/J. I. Fisiopatologia das arritmias e da morte súbita do chagásico. Rev. Ass. méd. bras., 29: 77-83, 1983.
20. TOWBIN, H.; STAEHLIN, T. & GORDON, J. - Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. nat. Acad. Sci. (Wash.), 76: 4350-4354, 1979.
21. VOLLER, A.; DRAPPER, C; BIDWELL, D.E. & BARTLETT, A. - Microplate enzyme-linked immunosorbent assay of Chagas' disease. Lancet, 1: 426-428, 1975.
22. WHO Control of Chagas' Disease. Report of a WHO Expert Committee. Wld. Hlth. Org. techn. Rep. Ser., (811):95, 1991.

Correspondence to:

Dr. Antonio R.L. Teixeira

Laboratório Multidisciplinar de Pesquisa em Doença de Chagas, Faculdade de Ciências da Saúde, Universidade de Brasília

Caixa Postal 04685

70919-970 Brasília, DF, Brasil

FAX: (061) 273-4645

Publication Dates

Publication in this collection
22 Sept 2006
Date of issue
June 1996

History

Accepted
07 May 1996
Received
30 Oct 1995

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

[1] 1. ALENCAR, J.E.; ALMEIDA, Y.M.; SILVA, Z.F.; PAIVA, S.A. & FONSECA, M.F. - Aspectos atuais do Calazar no Ceará. Rev. bras. Malar., 25: 27-53, 1974.

[2] 2. CAMARGO, M.E. - Fluorescent antibody test for the serodiagnosis of American Trypanosomiasis: technical modification employing preserved culture forms of Trypanosoma cruzi in a slide test. Rev. Inst. Med. trop. S. Paulo, 8: 227-234, 1966.

[3] 3. CAMARGO, M.E. & REBONATO, C. - Cross-reactivity in immunofluorescence test for Trypanosoma and Leishmania antibodies. A simple inhibition procedure to ensure specific results. Amer J. trop. Med. Hyg., 18: 500-505, 1969.

[4] 4. CHILLER, T.M.; SAMUDIO, M.A. & ZOULEK, G. - IgG antibody reactivity with Trypanosoma cruzi and Leishmania antigens in sera of patients with Chagas' disease and leishmaniasis. Amer. J. trop. Med. Hyg., 43: 249-259, 1990.

[5] 5. CUBA-CUBA, C.A.; MARSDEN, P.D.; BARRETO, A.C. et al. - Parasitologic and immunologic diagnosis of American (mucocutaneous) leishmaniasis. Bull. Pan Amer. Hlth. Org., 15: 249-259, 1981.

[6] 6. EVANS, D.A. Leishmania reference strains. Parasit. today, 1: 172-173, 1985.

[7] 7. GRIMALDI Jr., G.; TESHL, R.B. & MCMAHON-PRATT, D. - A review of the geographic distribution and epidemiology of leishmaniasis in the New World. Amer. J. trop. Med. Hyg., 41: 687-725, 1989.

[8] 8. HARITH, A.E.; KOLK, A.H.J.; MINTER GOEDBLOED, E.; KAGER, P.A. & KOLK, A.H.J. - Improvement of a direct agglutination test for field studies of visceral leishmaniasis. J. clin. Microbiol., 26: 1321-1325, 1988.

[9] 9. LAEMMLI, U.K. - Cleavage of structural proteins during assembly of the head of bacteriophage. Nature, 227: 680-685, 1970.

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

[11] 11. MALCHIODI, E.L.; CHIARAMONTE, M.G.; TARANTO, N.J.; ZWIRNER, N.W. & MARGNI, R.A. - Cross-reactivity studies and differential diagnosis of human infections caused by Trypanosoma cruzi and Leishmania spp; use of immunobloting and ELISA with a purified antigen (Ag163Bb). Clin. exp. Immunol., 97: 417-423, 1994.

[12] 12. MARSDEN, P.D. - Leishmaniasis. New Engl. J. Med., 300: 350-352, 1974.

[13] 13. MASUDA, A.; NASCIMENTO, S.F.; GUERRA, C.S.; PARANHOS, G.S. & FERREIRA, A.W. - Analysis of the specificity of human antibodies to antigens of Leishmania braziliensis braziliensis. Rev. Inst. Med. trop. S. Paulo, 31: 228-234, 1989.

[14] 14. MOTT, K.E.; LEHMAN Jr., J.S.; HOFF, R. et al. - The epidemiology and household distribution of seroreactivity to Trypanosoma cruzi in a rural community in northeast Brazil. Amer J. trop. Med. Hyg., 25: 552-562, 1976.

[15] 15. NEVA, F.A. & GAM, A. - A complement-fixing antigen from Trypanosoma cruzi grown in cell cultures. Amer. J. trop. Med. Hyg., 26: 37-46, 1977.

[16] 16. SHAW, J.J. & LAINSON, R. - Ecology and epidemiology: new world. In: PETERS, W. & KILLICK-KENDRICK, R., ed. The leishmaniasis in biology and medicine. London, Academic-Press, 1987. v. 1, p. 291-363.

[17] 17. TEIXEIRA, A.R.L. & PEREIRA, L.M. - Discrepâncias entre resultados de tręs reaçőes sorológicas empregadas para diagnóstico da Doença de Chagas. Rev. bras. Biol., 42: 789-795, 1981.

[18] 18. TEIXEIRA, A.R.L.; JUNQUEIRA Jr., L.F.; SOLÓRZANO, E. & ZAPPALA, M. - Doença de Chagas experimental em coelhos isogęnicos III/J. I. Fisiopatologia das arritmias e da morte súbita do chagásico. Rev. Ass. méd. bras., 29: 77-83, 1983.

[19] 20. TOWBIN, H.; STAEHLIN, T. & GORDON, J. - Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. nat. Acad. Sci. (Wash.), 76: 4350-4354, 1979.

[20] 21. VOLLER, A.; DRAPPER, C; BIDWELL, D.E. & BARTLETT, A. - Microplate enzyme-linked immunosorbent assay of Chagas' disease. Lancet, 1: 426-428, 1975.

[21] 22. WHO Control of Chagas' Disease. Report of a WHO Expert Committee. Wld. Hlth. Org. techn. Rep. Ser., (811):95, 1991.

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