This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Oliveira, T. R.
Right arrow Articles by Nascimento, A. L. T. O.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Oliveira, T. R.
Right arrow Articles by Nascimento, A. L. T. O.

 Previous Article  |  Next Article 

Clinical and Vaccine Immunology, November 2008, p. 1715-1722, Vol. 15, No. 11
1071-412X/08/$08.00+0     doi:10.1128/CVI.00214-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Evaluation of Leptospiral Recombinant Antigens MPL17 and MPL21 for Serological Diagnosis of Leptospirosis by Enzyme-Linked Immunosorbent Assays {triangledown}

Tatiane R. Oliveira,1 Mariana T. Longhi,1 Zenaide M. de Morais,2 Eliete C. Romero,3 Roberta M. Blanco,3 Karin Kirchgatter,4 Silvio A. Vasconcellos,2 and Ana L. T. O. Nascimento1,5*

Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, São Paulo, SP 05503-900, Brazil,1 Laboratório de Zoonoses Bacterianas do VPS, Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, São Paulo, Brazil,2 Divisão de Biologia Medica, Instituto Adolfo Lutz, São Paulo, Brazil,3 Núcleo de Estudos em Malária, Superintendência de Controle de Endemias/Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brazil,4 Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, USP, São Paulo, Brazil5

Received 9 June 2008/ Returned for modification 11 August 2008/ Accepted 6 September 2008

Leptospirosis is a zoonosis of multisystem involvement caused by pathogenic strains of the genus Leptospira. In the last few years, intensive studies aimed at the development of a vaccine have provided important knowledge about the nature of the immunological mechanisms of the host. The purpose of this study was to analyze the immune responses to two recombinant proteins, MPL17 and MPL21 (encoded by the genes LIC10765 and LIC13131, respectively) of Leptospira interrogans serovar Copenhageni in individuals during infection. The recombinant proteins were expressed in Escherichia coli as six-His tag fusion proteins and were purified from the soluble bacterial fraction by affinity chromatography with Ni2+-charged resin. The recombinant proteins were used to evaluate their ability to bind to immunoglobulin G (IgG) (and IgG subclass) or IgM antibodies in serum samples from patients in the early and convalescent phases of leptospirosis (n = 52) by enzyme-linked immunosorbent assays. The prevalences of total IgG antibodies against MPL17 and MPL21 were 38.5% and 21.2%, respectively. The titers achieved with MPL17 were statistically significantly higher than those obtained by the reference microscopic agglutination test. The specificity of the assay was estimated to be 95.5% for MPL17 and 80.6% for MPL21 when serum samples from individuals with unrelated febrile diseases and control healthy donors were tested. The proteins are conserved among Leptospira strains that cause human and animal diseases. MPL17 and MPL21 are most likely new surface proteins of leptospires, as revealed by liquid-phase immunofluorescence assays with living organisms. Our results demonstrate that these recombinant proteins are highly immunogenic and, when they are used together, might be useful as a means of diagnosing leptospirosis.

* Corresponding author. Mailing address: Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil, 1500, São Paulo, SP 05503-900, Brazil. Phone: (5511) 37220019. Fax: (5511) 37261505. E-mail: tabet{at}butantan.gov.br

{triangledown} Published ahead of print on 17 September 2008.

Clinical and Vaccine Immunology, November 2008, p. 1715-1722, Vol. 15, No. 11
1071-412X/08/$08.00+0     doi:10.1128/CVI.00214-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»