Identification of Strain-Specific B-cell Epitopes in Trypanosoma cruzi Using Genome-Scale Epitope Prediction and High-Throughput Immunoscreening with Peptide Arrays

Tiago Antônio de Oliveira Mendes, João Luís Reis Cunha, Rodrigo de Almeida Lourdes, Gabriela Flávia Rodrigues Luiz, Lucas Dhom Lemos, Ana Rita Rocha dos Santos, Antônia Cláudia Jácome da Câmara, Lúcia Maria da Cunha Galvão, Caryn Bern, Robert H Gilman, Ricardo Toshio Fujiwara, Ricardo Tostes Gazzinelli, Daniella Castanheira Bartholomeu

Research output: Contribution to journalArticle

Abstract

Background:The factors influencing variation in the clinical forms of Chagas disease have not been elucidated; however, it is likely that the genetics of both the host and the parasite are involved. Several studies have attempted to correlate the T. cruzi strains involved in infection with the clinical forms of the disease by using hemoculture and/or PCR-based genotyping of parasites from infected human tissues. However, both techniques have limitations that hamper the analysis of large numbers of samples. The goal of this work was to identify conserved and polymorphic linear B-cell epitopes of T. cruzi that could be used for serodiagnosis and serotyping of Chagas disease using ELISA.Methodology:By performing B-cell epitope prediction on proteins derived from pair of alleles of the hybrid CL Brener genome, we have identified conserved and polymorphic epitopes in the two CL Brener haplotypes. The rationale underlying this strategy is that, because CL Brener is a recent hybrid between the TcII and TcIII DTUs (discrete typing units), it is likely that polymorphic epitopes in pairs of alleles could also be polymorphic in the parental genotypes. We excluded sequences that are also present in the Leishmania major, L. infantum, L. braziliensis and T. brucei genomes to minimize the chance of cross-reactivity. A peptide array containing 150 peptides was covalently linked to a cellulose membrane, and the reactivity of the peptides was tested using sera from C57BL/6 mice chronically infected with the Colombiana (TcI) and CL Brener (TcVI) clones and Y (TcII) strain.Findings and Conclusions:A total of 36 peptides were considered reactive, and the cross-reactivity among the strains is in agreement with the evolutionary origin of the different T. cruzi DTUs. Four peptides were tested against a panel of chagasic patients using ELISA. A conserved peptide showed 95.8% sensitivity, 88.5% specificity, and 92.7% accuracy for the identification of T. cruzi in patients infected with different strains of the parasite. Therefore, this peptide, in association with other T. cruzi antigens, may improve Chagas disease serodiagnosis. Together, three polymorphic epitopes were able to discriminate between the three parasite strains used in this study and are thus potential targets for Chagas disease serotyping.

Original languageEnglish (US)
Article numbere2524
JournalPLoS Neglected Tropical Diseases
Volume7
Issue number10
DOIs
StatePublished - Oct 2013

Fingerprint

B-Lymphocyte Epitopes
Trypanosoma cruzi
Epitopes
Genome
Chagas Disease
Peptides
Parasites
Serotyping
Serologic Tests
Enzyme-Linked Immunosorbent Assay
Alleles
Leishmania major
Viral Tumor Antigens
Inbred C57BL Mouse
Cellulose
Haplotypes
Clone Cells
Genotype
Sensitivity and Specificity
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Infectious Diseases
  • Public Health, Environmental and Occupational Health
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

de Oliveira Mendes, T. A., Reis Cunha, J. L., de Almeida Lourdes, R., Rodrigues Luiz, G. F., Lemos, L. D., dos Santos, A. R. R., ... Bartholomeu, D. C. (2013). Identification of Strain-Specific B-cell Epitopes in Trypanosoma cruzi Using Genome-Scale Epitope Prediction and High-Throughput Immunoscreening with Peptide Arrays. PLoS Neglected Tropical Diseases, 7(10), [e2524]. https://doi.org/10.1371/journal.pntd.0002524

Identification of Strain-Specific B-cell Epitopes in Trypanosoma cruzi Using Genome-Scale Epitope Prediction and High-Throughput Immunoscreening with Peptide Arrays. / de Oliveira Mendes, Tiago Antônio; Reis Cunha, João Luís; de Almeida Lourdes, Rodrigo; Rodrigues Luiz, Gabriela Flávia; Lemos, Lucas Dhom; dos Santos, Ana Rita Rocha; da Câmara, Antônia Cláudia Jácome; da Cunha Galvão, Lúcia Maria; Bern, Caryn; Gilman, Robert H; Fujiwara, Ricardo Toshio; Gazzinelli, Ricardo Tostes; Bartholomeu, Daniella Castanheira.

In: PLoS Neglected Tropical Diseases, Vol. 7, No. 10, e2524, 10.2013.

Research output: Contribution to journalArticle

de Oliveira Mendes, TA, Reis Cunha, JL, de Almeida Lourdes, R, Rodrigues Luiz, GF, Lemos, LD, dos Santos, ARR, da Câmara, ACJ, da Cunha Galvão, LM, Bern, C, Gilman, RH, Fujiwara, RT, Gazzinelli, RT & Bartholomeu, DC 2013, 'Identification of Strain-Specific B-cell Epitopes in Trypanosoma cruzi Using Genome-Scale Epitope Prediction and High-Throughput Immunoscreening with Peptide Arrays', PLoS Neglected Tropical Diseases, vol. 7, no. 10, e2524. https://doi.org/10.1371/journal.pntd.0002524
de Oliveira Mendes, Tiago Antônio ; Reis Cunha, João Luís ; de Almeida Lourdes, Rodrigo ; Rodrigues Luiz, Gabriela Flávia ; Lemos, Lucas Dhom ; dos Santos, Ana Rita Rocha ; da Câmara, Antônia Cláudia Jácome ; da Cunha Galvão, Lúcia Maria ; Bern, Caryn ; Gilman, Robert H ; Fujiwara, Ricardo Toshio ; Gazzinelli, Ricardo Tostes ; Bartholomeu, Daniella Castanheira. / Identification of Strain-Specific B-cell Epitopes in Trypanosoma cruzi Using Genome-Scale Epitope Prediction and High-Throughput Immunoscreening with Peptide Arrays. In: PLoS Neglected Tropical Diseases. 2013 ; Vol. 7, No. 10.
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abstract = "Background:The factors influencing variation in the clinical forms of Chagas disease have not been elucidated; however, it is likely that the genetics of both the host and the parasite are involved. Several studies have attempted to correlate the T. cruzi strains involved in infection with the clinical forms of the disease by using hemoculture and/or PCR-based genotyping of parasites from infected human tissues. However, both techniques have limitations that hamper the analysis of large numbers of samples. The goal of this work was to identify conserved and polymorphic linear B-cell epitopes of T. cruzi that could be used for serodiagnosis and serotyping of Chagas disease using ELISA.Methodology:By performing B-cell epitope prediction on proteins derived from pair of alleles of the hybrid CL Brener genome, we have identified conserved and polymorphic epitopes in the two CL Brener haplotypes. The rationale underlying this strategy is that, because CL Brener is a recent hybrid between the TcII and TcIII DTUs (discrete typing units), it is likely that polymorphic epitopes in pairs of alleles could also be polymorphic in the parental genotypes. We excluded sequences that are also present in the Leishmania major, L. infantum, L. braziliensis and T. brucei genomes to minimize the chance of cross-reactivity. A peptide array containing 150 peptides was covalently linked to a cellulose membrane, and the reactivity of the peptides was tested using sera from C57BL/6 mice chronically infected with the Colombiana (TcI) and CL Brener (TcVI) clones and Y (TcII) strain.Findings and Conclusions:A total of 36 peptides were considered reactive, and the cross-reactivity among the strains is in agreement with the evolutionary origin of the different T. cruzi DTUs. Four peptides were tested against a panel of chagasic patients using ELISA. A conserved peptide showed 95.8{\%} sensitivity, 88.5{\%} specificity, and 92.7{\%} accuracy for the identification of T. cruzi in patients infected with different strains of the parasite. Therefore, this peptide, in association with other T. cruzi antigens, may improve Chagas disease serodiagnosis. Together, three polymorphic epitopes were able to discriminate between the three parasite strains used in this study and are thus potential targets for Chagas disease serotyping.",
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T1 - Identification of Strain-Specific B-cell Epitopes in Trypanosoma cruzi Using Genome-Scale Epitope Prediction and High-Throughput Immunoscreening with Peptide Arrays

AU - de Oliveira Mendes, Tiago Antônio

AU - Reis Cunha, João Luís

AU - de Almeida Lourdes, Rodrigo

AU - Rodrigues Luiz, Gabriela Flávia

AU - Lemos, Lucas Dhom

AU - dos Santos, Ana Rita Rocha

AU - da Câmara, Antônia Cláudia Jácome

AU - da Cunha Galvão, Lúcia Maria

AU - Bern, Caryn

AU - Gilman, Robert H

AU - Fujiwara, Ricardo Toshio

AU - Gazzinelli, Ricardo Tostes

AU - Bartholomeu, Daniella Castanheira

PY - 2013/10

Y1 - 2013/10

N2 - Background:The factors influencing variation in the clinical forms of Chagas disease have not been elucidated; however, it is likely that the genetics of both the host and the parasite are involved. Several studies have attempted to correlate the T. cruzi strains involved in infection with the clinical forms of the disease by using hemoculture and/or PCR-based genotyping of parasites from infected human tissues. However, both techniques have limitations that hamper the analysis of large numbers of samples. The goal of this work was to identify conserved and polymorphic linear B-cell epitopes of T. cruzi that could be used for serodiagnosis and serotyping of Chagas disease using ELISA.Methodology:By performing B-cell epitope prediction on proteins derived from pair of alleles of the hybrid CL Brener genome, we have identified conserved and polymorphic epitopes in the two CL Brener haplotypes. The rationale underlying this strategy is that, because CL Brener is a recent hybrid between the TcII and TcIII DTUs (discrete typing units), it is likely that polymorphic epitopes in pairs of alleles could also be polymorphic in the parental genotypes. We excluded sequences that are also present in the Leishmania major, L. infantum, L. braziliensis and T. brucei genomes to minimize the chance of cross-reactivity. A peptide array containing 150 peptides was covalently linked to a cellulose membrane, and the reactivity of the peptides was tested using sera from C57BL/6 mice chronically infected with the Colombiana (TcI) and CL Brener (TcVI) clones and Y (TcII) strain.Findings and Conclusions:A total of 36 peptides were considered reactive, and the cross-reactivity among the strains is in agreement with the evolutionary origin of the different T. cruzi DTUs. Four peptides were tested against a panel of chagasic patients using ELISA. A conserved peptide showed 95.8% sensitivity, 88.5% specificity, and 92.7% accuracy for the identification of T. cruzi in patients infected with different strains of the parasite. Therefore, this peptide, in association with other T. cruzi antigens, may improve Chagas disease serodiagnosis. Together, three polymorphic epitopes were able to discriminate between the three parasite strains used in this study and are thus potential targets for Chagas disease serotyping.

AB - Background:The factors influencing variation in the clinical forms of Chagas disease have not been elucidated; however, it is likely that the genetics of both the host and the parasite are involved. Several studies have attempted to correlate the T. cruzi strains involved in infection with the clinical forms of the disease by using hemoculture and/or PCR-based genotyping of parasites from infected human tissues. However, both techniques have limitations that hamper the analysis of large numbers of samples. The goal of this work was to identify conserved and polymorphic linear B-cell epitopes of T. cruzi that could be used for serodiagnosis and serotyping of Chagas disease using ELISA.Methodology:By performing B-cell epitope prediction on proteins derived from pair of alleles of the hybrid CL Brener genome, we have identified conserved and polymorphic epitopes in the two CL Brener haplotypes. The rationale underlying this strategy is that, because CL Brener is a recent hybrid between the TcII and TcIII DTUs (discrete typing units), it is likely that polymorphic epitopes in pairs of alleles could also be polymorphic in the parental genotypes. We excluded sequences that are also present in the Leishmania major, L. infantum, L. braziliensis and T. brucei genomes to minimize the chance of cross-reactivity. A peptide array containing 150 peptides was covalently linked to a cellulose membrane, and the reactivity of the peptides was tested using sera from C57BL/6 mice chronically infected with the Colombiana (TcI) and CL Brener (TcVI) clones and Y (TcII) strain.Findings and Conclusions:A total of 36 peptides were considered reactive, and the cross-reactivity among the strains is in agreement with the evolutionary origin of the different T. cruzi DTUs. Four peptides were tested against a panel of chagasic patients using ELISA. A conserved peptide showed 95.8% sensitivity, 88.5% specificity, and 92.7% accuracy for the identification of T. cruzi in patients infected with different strains of the parasite. Therefore, this peptide, in association with other T. cruzi antigens, may improve Chagas disease serodiagnosis. Together, three polymorphic epitopes were able to discriminate between the three parasite strains used in this study and are thus potential targets for Chagas disease serotyping.

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