Leishmania braziliensis: isolation of carbohydrate-containing antigen and possibility of its use in the immunodiagnosis of American Cutaneous Leishmaniasis

Leishmania braziliensis: isolamento de antígeno contendo carboidrato e a possibilidade de sua aplicação no imunodiagnóstico da Leishmaniose Tegumentar Americana

T. G. V. Silveira C Kemmelmeier About the authors

Abstracts

Leishmania braziliensis is a causative agent of American Cutaneous Leishmaniasis (ACL). The 034-JCG strain, isolated from a patient from the northern region of Paraná State, Brazil, was cultivated in Blood Agar Base medium, lyophilized and submitted to phenol-water extraction. The extract was treated with RNase I. The carbohydrate containing-antigen (Ag-CHO) was immunogenic to rabbits and showed at least a fraction with some negative charge at pH 8.2. This antigen showed cross-reactivity with the phenol-water extract of the growth medium used for the culture of promastigotes and with the surface antigens of promastigotes. Its composition is: 24.3% of total sugars, from which 11.2% of galactose, 7.5% of mannose and 5.6% of ribose. Protein content was 5.4% and phosphate 18.5%. The antigenic activity was maintained after: repeated freezing-thawing; lyophilization; heating at 100ºC for 30 minutes; treatment with RNase, trichloroacetic acid and sodium metaperiodate. The precipitin line obtained is Periodic Acid Schiff positive. The application of the Ag-CHO in counterimmunoelectrophoresis reaction for the immunodiagnosis of ACL showed 60% sensitivity, and no cross-reaction with the five sera of Chagas' disease patients tested. The use of this antigen in a more sensitive technique, with more samples of sera, may improve these results.

Leishmania braziliensis; Carbohydrate-containing antigen; Immunodiagnosis


A Leishmania braziliensis é um dos agentes causadores da Leishmaniose Tegumentar Americana (LTA). A cepa 034-JCG, isolada de paciente da região norte do estado do Paraná, Brasil, foi cultivada em meio Blood Agar Base, liofilizada e submetida a extração com fenol-água e o extrato obtido foi tratado com RNase I. O antígeno contendo carboidrato (Ag-CHO) mostrou-se imunogênico para coelhos e apresentou pelo menos uma fração com poucas cargas negativas em pH 8,2. Este antígeno apresenta reação cruzada com extrato fenólico do meio de cultura usado para o cultivo de promastigotas e com antígenos de superfície de promastigotas. É composto por 24,3% de açúcares totais, dos quais 11,2% de galactose, 7,5% de manose e 5,6% de ribose. As proteínas foram estimadas em 5,4% e fosfato em 18,5%. A atividade antigênica foi mantida após: congelamentos e descongelamentos repetidos; liofilização; aquecimento a 100ºC durante 30 minutos; tratamento com RNase, ácido tricloroacético e periodato de sódio. As linhas obtidas nas reações de precipitação coram-se pelo Ácido Periódico de Schiff. A utilização do Ag-CHO na reação de contraimunoeletroforese para o imunodiagnóstico de LTA mostrou 60% de sensibilidade, e nenhuma reação cruzada com os 5 soros de pacientes com Doença de Chagas testados. O uso deste antígeno com técnicas mais sensíveis, e com um maior número de amostras poderá melhorar estes resultados.


DIAGNOSTIC STUDIES

Leishmania braziliensis: isolation of carbohydrate-containing antigen and possibility of its use in the immunodiagnosis of American Cutaneous Leishmaniasis

Leishmania braziliensis: isolamento de antígeno contendo carboidrato e a possibilidade de sua aplicação no imunodiagnóstico da Leishmaniose Tegumentar Americana

T. G. V. SilveiraI; C. KemmelmeierII

IDepartamento de Análises Clinicas - Universidade Estadual de Maringá. Maringá, Pr., Brasil

IIDepartamento de Bioquímica - Universidade Estadual de Maringá. Maringá, Pr., Brasil

Correspondence to

SUMMARY

Leishmania braziliensis is a causative agent of American Cutaneous Leishmaniasis (ACL). The 034-JCG strain, isolated from a patient from the northern region of Paraná State, Brazil, was cultivated in Blood Agar Base medium, lyophilized and submitted to phenol-water extraction. The extract was treated with RNase I. The carbohydrate containing-antigen (Ag-CHO) was immunogenic to rabbits and showed at least a fraction with some negative charge at pH 8.2. This antigen showed cross-reactivity with the phenol-water extract of the growth medium used for the culture of promastigotes and with the surface antigens of promastigotes. Its composition is: 24.3% of total sugars, from which 11.2% of galactose, 7.5% of mannose and 5.6% of ribose. Protein content was 5.4% and phosphate 18.5%. The antigenic activity was maintained after: repeated freezing-thawing; lyophilization; heating at 100ºC for 30 minutes; treatment with RNase, trichloroacetic acid and sodium metaperiodate. The precipitin line obtained is Periodic Acid Schiff positive. The application of the Ag-CHO in counterimmunoelectrophoresis reaction for the immunodiagnosis of ACL showed 60% sensitivity, and no cross-reaction with the five sera of Chagas' disease patients tested. The use of this antigen in a more sensitive technique, with more samples of sera, may improve these results.

Keywords:Leishmania braziliensis; Carbohydrate-containing antigen; Immunodiagnosis.

RESUMO

A Leishmania braziliensis é um dos agentes causadores da Leishmaniose Tegumentar Americana (LTA). A cepa 034-JCG, isolada de paciente da região norte do estado do Paraná, Brasil, foi cultivada em meio Blood Agar Base, liofilizada e submetida a extração com fenol-água e o extrato obtido foi tratado com RNase I. O antígeno contendo carboidrato (Ag-CHO) mostrou-se imunogênico para coelhos e apresentou pelo menos uma fração com poucas cargas negativas em pH 8,2. Este antígeno apresenta reação cruzada com extrato fenólico do meio de cultura usado para o cultivo de promastigotas e com antígenos de superfície de promastigotas. É composto por 24,3% de açúcares totais, dos quais 11,2% de galactose, 7,5% de manose e 5,6% de ribose. As proteínas foram estimadas em 5,4% e fosfato em 18,5%. A atividade antigênica foi mantida após: congelamentos e descongelamentos repetidos; liofilização; aquecimento a 100ºC durante 30 minutos; tratamento com RNase, ácido tricloroacético e periodato de sódio. As linhas obtidas nas reações de precipitação coram-se pelo Ácido Periódico de Schiff. A utilização do Ag-CHO na reação de contraimunoeletroforese para o imunodiagnóstico de LTA mostrou 60% de sensibilidade, e nenhuma reação cruzada com os 5 soros de pacientes com Doença de Chagas testados. O uso deste antígeno com técnicas mais sensíveis, e com um maior número de amostras poderá melhorar estes resultados.

Full text available only in PDF format.

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ACKNOWLEDGMENT

The authors wish to thank Dr. Adelar Bracht for the English review of the manuscript.

  • Correspondence to:
    T. G. V. Silveira
    Lab. Imunologia Clínica, Departamento de Análises Clínicas - Universidade Estadual de Maringá
    Av. Colombo, 5790
    87.020-900 Maringá, Pr., Brasil
  • Recebido para publicação em 07/06/1994.

    Aceito para publicação em 02/05/1995.

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    Correspondence to: T. G. V. Silveira Lab. Imunologia Clínica, Departamento de Análises Clínicas - Universidade Estadual de Maringá Av. Colombo, 5790 87.020-900 Maringá, Pr., Brasil

    Publication Dates

    • Publication in this collection
      21 Sept 2006
    • Date of issue
      June 1995

    History

    • Accepted
      02 May 1995
    • Received
      07 June 1994
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