Lytic antibodies elicited by Trypanosoma cruzi infection recognize epitopes present on both bloodstream trypomastigote and epimastigote forms of parasite

Takehara, Harumi A.; Cardoso, Diva F.; Silva, Ana M.M. da; Mota, I.

doi:10.1590/S0036-46651988000500004

Abstracts

Sera of Chaga's disease patients containing anti-T. cruzi lytic antibodies were submitted to affinity chromatography using Sepharose 4B conjugated with antigen extracted from epimasiigote or trypomasiigote forms of the parasite. Epimastigotes were obtained from culture at the exponential growth phase and the trypomastigotes from blood of infected and immunosuppressed mice. Antigen of both parasite forms was obtained by sonication of the parasites followed by centrifugation. Both antigens were then conjugated to activated Sepharose 4B. Affinity chromatography was performed by passing sera from chagasic patients through an immunoadsorbent column containing either epimasiigote or trypomasiigote antigens. Antibodies bound to the column were eluted with cold 0,2 M glycine buffer pH 2,8. The eluted antibodies were analysed regarding their isotype and lytic activity. The results showed that anti-T. cruzi lytic antibodies present in sera from chagasic patients are mainly located in the IgG isotype and recognize epitopes present in both trypomasiigote and epimastigote forms. A brief report of this work has already been published12.

Lytic antibody; Trypanosoma cruzi

Soro de pacientes com doença de Chagas na fase crônica foram submetidos a cromatografia de afinidade com Sepharose 4B conjugada com um extrato antigênico obtido de formas epimastigotas ou tripomastigotas de T. cruzi: os epimastigotas foram obtidos de cultura na fase exponencial de crescimento e os tripomastigotas de sangue de camundongos infectados e imunossuprimidos. Os antígenos de ambas formas parasitárias foram obtidos por tratamento dos parasitas por ultra-som, seguido de centrifugação. A cromatografia de afinidade foi feita passando-se os soros chagásicos através de uma coluna de imunoadsorvente contendo antígenos de epimastigotas ou tripomastigotas. Os anticorpos foram eluídos da coluna com tampão glicina 0,2 M pH 2,8 a 4°C. Os anticorpos eluidos foram analisados quanto ao seu isotipo e atividade lítica. Os resultados mostraram que os anticorpos anti-T. cruzi com atividade lítica presentes em soros chagásicos estão localizados no isotipo IgG e reconhecem epitopos presentes tanto nos tripomastigotas quanto nos epimastigotas.

ORIGINAL ARTICLES

Lytic antibodies elicited by Trypanosoma cruzi infection recognize epitopes present on both bloodstream trypomastigote and epimastigote forms of parasite

Anticorpos líticos induzidos por infecção pelo Trypanosoma cruzi reconhecem epitopos presentes nas formas tripomastigotas e epimastigotas do parasita

Harumi A. Takehara; Diva F. Cardoso; Ana M.M. da Silva; I. Mota

Centro de Pesquisa e Formação em Imunologia da OMS/OPS. Instituto Butantan, São Paulo, Brasil

^{Address for correspondence} Address for correspondence: Dra. Harumi A. Takehara Centro de Pesquisa e Formação em Imunologia da OMS/OPS Instituto Butantan Av. Dr. Vital Brasil, 1500 Caixa Postal 65 CEP 05504 São Paulo, SP., Brasil

SUMMARY

Sera of Chaga's disease patients containing anti-T. cruzi lytic antibodies were submitted to affinity chromatography using Sepharose 4B conjugated with antigen extracted from epimasiigote or trypomasiigote forms of the parasite. Epimastigotes were obtained from culture at the exponential growth phase and the trypomastigotes from blood of infected and immunosuppressed mice. Antigen of both parasite forms was obtained by sonication of the parasites followed by centrifugation. Both antigens were then conjugated to activated Sepharose 4B. Affinity chromatography was performed by passing sera from chagasic patients through an immunoadsorbent column containing either epimasiigote or trypomasiigote antigens. Antibodies bound to the column were eluted with cold 0,2 M glycine buffer pH 2,8. The eluted antibodies were analysed regarding their isotype and lytic activity. The results showed that anti-T. cruzi lytic antibodies present in sera from chagasic patients are mainly located in the IgG isotype and recognize epitopes present in both trypomasiigote and epimastigote forms. A brief report of this work has already been published¹².

Key words: Lytic antibody; Trypanosoma cruzi

RESUMO

Soro de pacientes com doença de Chagas na fase crônica foram submetidos a cromatografia de afinidade com Sepharose 4B conjugada com um extrato antigênico obtido de formas epimastigotas ou tripomastigotas de T. cruzi: os epimastigotas foram obtidos de cultura na fase exponencial de crescimento e os tripomastigotas de sangue de camundongos infectados e imunossuprimidos. Os antígenos de ambas formas parasitárias foram obtidos por tratamento dos parasitas por ultra-som, seguido de centrifugação.

A cromatografia de afinidade foi feita passando-se os soros chagásicos através de uma coluna de imunoadsorvente contendo antígenos de epimastigotas ou tripomastigotas. Os anticorpos foram eluídos da coluna com tampão glicina 0,2 M pH 2,8 a 4°C. Os anticorpos eluidos foram analisados quanto ao seu isotipo e atividade lítica. Os resultados mostraram que os anticorpos anti-T. cruzi com atividade lítica presentes em soros chagásicos estão localizados no isotipo IgG e reconhecem epitopos presentes tanto nos tripomastigotas quanto nos epimastigotas.

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ACKNOWLEDGMENTS

This work was supported by the Conselho Nacional de Pesquisas, Brazil (process 40.0695/85). We thank Dr. Rodolfo Pereira Mendes for the generous supply of sera from chagasic patients and Mr. Carlos Jared for technical assistance.

Recebido para publicação em 16/3/1988.

1. ARAÚJO, F.G. & REMINGTON, J.S. - Characterization of stages and strain of Trypanosoma cruzi by analysis of cell membrane components. J. Immunol., 127: 855-859, 1981.
2. CHAO, D. & DUSANIC, D.G. - Monoclonal antibodies to metacyclic stage antigens of Trypanosoma cruzi Amer. J. trop. Med. Hyg.,34: 694-701, 1985.
3. KIERSZENBAUM, F. - Cross-reactivity of lytic antibodies against blood forms of Trypanosoma cruzi J. Parasit., 62: 134-135, 1976.
4. KIPNIS, T.L.; SUCUPIRA, M. & DIAS DA SILVA; W. - Transformadon of Trypanosoma cruzi trypomastigote bloodstream forms by immune IgM and its Fatyi fragment into activators of the alternative complement pathway. Braz. J. med. biol. Res., 20: 105-114, 1987.
5. KRETTLI, A.U. - Efeito de anticorpos e do complemento sobre tripomastigotas sanguíneos de camundongos infectados com Trypanosoma cruzi. Belo Horizonte, 1978. (Tese de doutoramento - Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais).
6. KRETTLI, A.U. & BRENER, Z. - Resistance against Trypanosoma cruzi associated to anti-living trypomastigote antibodies. J. Immunol., 128: 2009-2012, 1982.
7. KRETTLI, A.U.; WEISS-CARRINGTON, P. & NUSSENZWEIG, R.S. - Membrane-bound antibodies to bloodstream Trypanosoma cruzi in mice: strain differences in susceptibility to complement-mediated lysis. Clin. exp. Immunol., 37: 416-423, 1979.
8. ROMEIRO, S.A.; TAKEHARA, H.A. & MOTA, I. - Isotype of lytic antibodies in serum of Chagas' disease patients. Clin. exp. Immunol., 55: 413-418, 1984.
9. SEGURA, E.L.; BUA, J.; ROSENSTEIN DE CAMPANINI, A.; SUBIAS, E.; ESTEVA, M.; MORENO, M. & RUIZ, A.M. - Monoclonal antibodies against the flagellar fraction of epimastigotes of Trypanosoma cruzi: complement - mediated lytic activity against trypomastigotes and passive immunoprotection in mice. Immunol. Lett., 13: 165-171, 1986.
10. SNARY, D. - Cell surface glycoproteins of Trypanosoma cruzi: protective immunity in mice and antibody levels in human chagasic sera. Trans. roy. Soc. trop. Med. Hyg., 77: 126-129, 1983.
11. SNARY, D. & HUDSON, L. - Trypanosoma cruzi cell surface proteins: identification and isolation of one major glycoprotein. FEBS Letters, 100:166-170, 1979.
12. TAKEHARA, H.A.; CARDOSO, D.F. & MOTA, I. - Isolation of lytic antibodies with T. cruzi antigens. In: INTERNATIONAL CONGRESS OF IMMUNOLOGY, 6., Toronto, Canada, 1986. Abstracts, p. 649.
13. TEIXEIRA, M.M.G. & YOSHIDA, N. - Stagespecific surface antigens of metacyclic trypomastigote of Trypanosoma cruzi identified by monoclonal antibodies. Molec. Biochem. Parasit., 18: 271-282, 1986.
14. TOWBIN, H.; STAHELIN, T. & GORDON, J. - Eletrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. nat. Acad. Sci. (Wash.), 76: 4350-4354, 1979.
15. YOSHIDA, N. - Surface antigens of metacyclic trypomastigote of Trypanosoma cruzi Infect. Immun., 40: 836-839, 1983.
16. YOSHIDA, N. - Trypanosoma cruzi: recognition of trypomastigote surface antigens by lytic antisera from mice resistant to acute infection. Exp. Parasit., 61: 184-191, 1986.

Address for correspondence:

Dra. Harumi A. Takehara

Centro de Pesquisa e Formação em Imunologia da OMS/OPS

Instituto Butantan

Av. Dr. Vital Brasil, 1500

Caixa Postal 65 CEP 05504 São Paulo, SP., Brasil

Publication Dates

Publication in this collection
17 Feb 2011
Date of issue
Oct 1988

History

Received
16 Mar 1988

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

[1] 1. ARAÚJO, F.G. & REMINGTON, J.S. - Characterization of stages and strain of Trypanosoma cruzi by analysis of cell membrane components. J. Immunol., 127: 855-859, 1981.

[2] 2. CHAO, D. & DUSANIC, D.G. - Monoclonal antibodies to metacyclic stage antigens of Trypanosoma cruzi Amer. J. trop. Med. Hyg.,34: 694-701, 1985.

[3] 3. KIERSZENBAUM, F. - Cross-reactivity of lytic antibodies against blood forms of Trypanosoma cruzi J. Parasit., 62: 134-135, 1976.

[4] 4. KIPNIS, T.L.; SUCUPIRA, M. & DIAS DA SILVA; W. - Transformadon of Trypanosoma cruzi trypomastigote bloodstream forms by immune IgM and its Fatyi fragment into activators of the alternative complement pathway. Braz. J. med. biol. Res., 20: 105-114, 1987.

[5] 5. KRETTLI, A.U. - Efeito de anticorpos e do complemento sobre tripomastigotas sanguíneos de camundongos infectados com Trypanosoma cruzi. Belo Horizonte, 1978. (Tese de doutoramento - Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais).

[6] 6. KRETTLI, A.U. & BRENER, Z. - Resistance against Trypanosoma cruzi associated to anti-living trypomastigote antibodies. J. Immunol., 128: 2009-2012, 1982.

[7] 7. KRETTLI, A.U.; WEISS-CARRINGTON, P. & NUSSENZWEIG, R.S. - Membrane-bound antibodies to bloodstream Trypanosoma cruzi in mice: strain differences in susceptibility to complement-mediated lysis. Clin. exp. Immunol., 37: 416-423, 1979.

[8] 8. ROMEIRO, S.A.; TAKEHARA, H.A. & MOTA, I. - Isotype of lytic antibodies in serum of Chagas' disease patients. Clin. exp. Immunol., 55: 413-418, 1984.

[9] 9. SEGURA, E.L.; BUA, J.; ROSENSTEIN DE CAMPANINI, A.; SUBIAS, E.; ESTEVA, M.; MORENO, M. & RUIZ, A.M. - Monoclonal antibodies against the flagellar fraction of epimastigotes of Trypanosoma cruzi: complement - mediated lytic activity against trypomastigotes and passive immunoprotection in mice. Immunol. Lett., 13: 165-171, 1986.

[10] 10. SNARY, D. - Cell surface glycoproteins of Trypanosoma cruzi: protective immunity in mice and antibody levels in human chagasic sera. Trans. roy. Soc. trop. Med. Hyg., 77: 126-129, 1983.

[11] 11. SNARY, D. & HUDSON, L. - Trypanosoma cruzi cell surface proteins: identification and isolation of one major glycoprotein. FEBS Letters, 100:166-170, 1979.

[12] 12. TAKEHARA, H.A.; CARDOSO, D.F. & MOTA, I. - Isolation of lytic antibodies with T. cruzi antigens. In: INTERNATIONAL CONGRESS OF IMMUNOLOGY, 6., Toronto, Canada, 1986. Abstracts, p. 649.

[13] 13. TEIXEIRA, M.M.G. & YOSHIDA, N. - Stagespecific surface antigens of metacyclic trypomastigote of Trypanosoma cruzi identified by monoclonal antibodies. Molec. Biochem. Parasit., 18: 271-282, 1986.

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

[15] 15. YOSHIDA, N. - Surface antigens of metacyclic trypomastigote of Trypanosoma cruzi Infect. Immun., 40: 836-839, 1983.

[16] 16. YOSHIDA, N. - Trypanosoma cruzi: recognition of trypomastigote surface antigens by lytic antisera from mice resistant to acute infection. Exp. Parasit., 61: 184-191, 1986.

Brasil

Brasil

Lytic antibodies elicited by Trypanosoma cruzi infection recognize epitopes present on both bloodstream trypomastigote and epimastigote forms of parasite

Anticorpos líticos induzidos por infecção pelo Trypanosoma cruzi reconhecem epitopos presentes nas formas tripomastigotas e epimastigotas do parasita

Abstracts

Publication Dates

History