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Development of a chimeric protein based on a proteomic approach for the serological diagnosis of human tegumentary leishmaniasis

  • Applied Genetics and Molecular Biotechnology
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Abstract

Leishmania braziliensis is responsible for most cases of human tegumentary leishmaniasis (HTL) and has caused a wide range of clinical manifestations, including cutaneous (CL) and mucosal leishmaniasis (ML). The diagnosis is based on criteria that consider epidemiological data, clinical findings, and laboratory tests and is hard to establish. For laboratory tests, none of the assays available can be considered gold standards for disease detection. In addition, the Montenegro skin test, essential to supporting infectologists in the clinical management of the disease, is no longer available in Brazil. Thus, the aim of this study was to develop new targets to be used in diagnostic tests for HTL. In the first step, we carried out two-dimensional gel electrophoresis, followed by mass spectrometry, combined with heat map analysis and immunoproteomics approach, and disclosed eight proteins expressed in the amastigote stage specifically recognized by serum from CL and ML patients. A chimeric protein was designed based on the combination of thirteen linear B-cell epitopes, identified by immunoinformatics analysis, from L. braziliensis proteins. Our results showed that the strategy used in this work was successful in developing an antigen to be used in immunological assays (100.0% sensitivity and specificity) in the detection of HTL cases and in comparison with results obtained from an ELISA using soluble L. braziliensis antigen (SLb-Antigen) and immunofluorescence assay (Bio-Manguinhos/FIOCRUZ). The present technology opens the door for its use in field exams by means of an immunochromatographic test, which will be even more helpful in regions without laboratory structures.

Key points

• Rational strategy to develop antigens.

• Integration between immunoproteomic and immunoinformatics analysis.

• Chimeric protein shows high performance in HTL diagnosis.

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Data availability

The authors declare that (the/all other) data supporting the findings of this study are available within the article (and its supplementary information files).

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Acknowledgements

MCD, TAOM, EAFC, and DM-S wish to thank Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brazil, for their fellowships.

Funding

This work was supported by grants from PRPq/UFMG (87–05/2016), CAPES (23038.004862/2015–74), FAPEMIG (APQ-03599–18 and APQ-03821–18), and CNPq (309939/2016–0, 408408/2016–2, 311426/2019–0, and 313070/2018–0).

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Authors and Affiliations

  1. Programa de Pós-Graduação Em Ciências da Saúde: Infectologia E Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil

    Guilherme Caetano Garcia, Ana Maria Ravena Severino Carvalho, Mariana Costa Duarte, Matheus Fernandes Costa e Silva, Fernanda Alvarenga Cardoso Medeiros, Eduardo Antonio Ferraz Coelho, Dênia Monteiro de Moura Franco, Denise Utsch Gonçalves & Daniel Menezes-Souza

  2. Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil

    Mariana Costa Duarte, Eduardo Antonio Ferraz Coelho & Daniel Menezes-Souza

  3. Departamento de Bioquímica E Biologia Molecular, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil

    Tiago Antônio de Oliveira Mendes

Authors
  1. Guilherme Caetano Garcia
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  2. Ana Maria Ravena Severino Carvalho
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  3. Mariana Costa Duarte
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  4. Matheus Fernandes Costa e Silva
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  6. Eduardo Antonio Ferraz Coelho
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  8. Denise Utsch Gonçalves
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  9. Tiago Antônio de Oliveira Mendes
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  10. Daniel Menezes-Souza
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Contributions

DM-S and TAOM conceived and designed research. GCG, AMRS, MCD, MFCS, FACM, and DMMF conducted experiments. DM-S, EAFC, and DTG contributed new reagents or analytical tools. DM-S, TAOM, GCG, and MCD performed result interpretation and data analysis. GCG and DM-S wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Daniel Menezes-Souza.

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Ethical approval

The use of human samples was approved by the Ethics Committee of the Federal University of Minas Gerais (UFMG), Belo Horizonte, Brazil (protocol CAAE-32343114.9.0000.5149). All patients received an individual copy of the study policy, which was reviewed by an independent person, and all participants signed an informed consent form, in Portuguese, before blood collection. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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The authors declare no competing interests.

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Garcia, G.C., Carvalho, A.M.R.S., Duarte, M.C. et al. Development of a chimeric protein based on a proteomic approach for the serological diagnosis of human tegumentary leishmaniasis. Appl Microbiol Biotechnol 105, 6805–6817 (2021). https://doi.org/10.1007/s00253-021-11518-1

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