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.
Texto completo disponível apenas em PDF.
Full text available only in PDF format.
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.
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Publication Dates
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Publication in this collection
17 Feb 2011 -
Date of issue
Oct 1988
History
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Received
16 Mar 1988