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A RECOMBINANT CYSTEINE PROTEINASE FROM LEISHMANIA (LEISHMANIA) CHAGASI SUITABLE FOR SERODIAGNOSIS OF AMERICAN VISCERAL LEISHMANIASIS

A recombinant protein, rLdccys1, which was produced by expression of the gene encoding a 30 kDa cysteine proteinase from Leishmania (Leishmania) chagasi, was used for detection of antibodies in sera from patients with active visceral leishmaniasis (VL) in enzyme-linked immunosorbent assays. Analysis of the predicted amino acid sequence of rLdccys1 showed that it contains all the characteristics of a cysteine proteinase. The ability of the protein to react with sera from humans with VL was also shown by Western blotting. The sensitivity for detection of specific antibodies to L. (L.) chagasi bodies using rLdccys1, L. (L.) chagasi promastigote lysates, and amastigote lysates was 80%, 98%, and 99%, respectively. No cross-reactivity between rLdccys1 and Chagas disease was observed, and there was little positive reactivity with sera from patients with cutaneous leishmaniasis and tuberculosis, compared with promastigote and amastigote extracts. Our findings indicate that rLdccys1 from L. (L.) chagasi constitutes a potential tool for the diagnosis of American VL.

Received March 4, 2004. Accepted for publication April 24, 2004.

Acknowledgments: We are grateful to Drs. Maurício Martins Rodrigues, Miriam Dorta, and Reinaldo Salomão for providing the human sera. We also thank Dr. Lindsay Pirrit for a critical reading of the manuscript.

Financial support: This work was supported by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior and the Fundação de Amparo à Pesquisa do Estado de Sao Paulo of Brazil.

Authors’ addresses: Suzana de Souza Dias, Simone Katz, Márcia Regina Machado dos Santos, and Clara Lúcia Barbiéri, Department of Microbiology, Immunology and Parasitology, Universidade Federal de São Paulo, Escola Paulista de Medicina, Rua Botucatu, 862, 60 Andar, 04023-062, São Paulo, SP, Brazil, Telephone: 55-11-5576-4532, Fax: 55-11-5571-1095, E-mails: suzi.dias{at}bol.com.br, simone.epm{at}uol.com.br, mrmachados{at}ecb.epm.br, and barbiericl{at}ecb.epm.br. Paulo Henrique da Costa Pinheiro, Associação de Ensino Superior e Tecnológico do Piauí, Rua Vitorino Ortiges Fernandes, 6123, 64057-100, Teresina, Piauí, Brazil, Telephone: 55-86-231-0700, Fax: 55-86- 231-0284, E-mail: ph.pinhe{at}bol.com.br.

Reprint requests: Clara Lúcia Barbiéri, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Botucatu, 862, 60 Andar, 04023-062, São Paulo, SP, Brazil.

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