Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications
Visceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have b...
| Autores: | , , , , , , , , , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Fecha de publicación: | 2014 |
| País: | España |
| Institución: | Universidad Autónoma de Madrid |
| Repositorio: | Biblos-e Archivo. Repositorio Institucional de la UAM |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.uam.es:10486/662771 |
| Acceso en línea: | http://hdl.handle.net/10486/662771 https://dx.doi.org/10.1128/CVI.00583-13 |
| Access Level: | acceso abierto |
| Palabra clave: | Leishmaniasis Leishmania infantum Potential serodiagnosis applications Biología y Biomedicina / Biología |
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Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applicationsCosta, Lourena EmanueleLima, Mayara Ingrid SousaChávez-Fumagalli, Miguel AngelMenezes-Souza, DanielMartins, Vivian TamiettiDuarte, Mariana CostaLage, Paula SousaLopes, Eliane Gonçalves PaivaLage, Daniela P.Ribeiro, Tatiana GomesAndrade, Pedro H RDe Magalhães Soares, Danielle FerreiraSoto Álvarez, ManuelTavares, Carlos Alberto PereiraGoulart, Luiz RicardoCoelho, Eduardo Anton̂io FerrazLeishmaniasisLeishmania infantumPotential serodiagnosis applicationsBiología y Biomedicina / BiologíaVisceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy and Trypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected with Leishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n 31) compared to those from vaccinated dogs (n 21), experimentally infected dogs with cross-reactive parasites (n 23), and healthy controls (n 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no crossreactivity with T. cruzi- or Ehrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes of L. infantum antigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programsThis work was supported by grants from the Pró-Reitoria de Pesquisa of UFMG (supported 03/2013), the Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (INCT Nano-Biofar), Rede Nanobiotec/Brasil-UFU (CAPES), PRONEX-FAPEMIG (APQ-01019- 09), FAPEMIG (APQ-00496-11 and APQ-00819-12), and CNPq (APQ- 472090/2011-9 and APQ-482976/2012-8). E.A.F.C. and L.R.G. are recipients of grants from CNPq. M.A.C.-F. is the recipient of a grant from FAPEMIG/CAPESAmerican Society for Microbiology.Departamento de Biología MolecularFacultad de CienciasCentro de Biología Molecular Severo Ochoa (CBM)20142014-01-01research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10486/662771https://dx.doi.org/10.1128/CVI.00583-13reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/6627712026-06-23T12:46:27Z |
| dc.title.none.fl_str_mv |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications |
| title |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications |
| spellingShingle |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications Costa, Lourena Emanuele Leishmaniasis Leishmania infantum Potential serodiagnosis applications Biología y Biomedicina / Biología |
| title_short |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications |
| title_full |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications |
| title_fullStr |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications |
| title_full_unstemmed |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications |
| title_sort |
Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications |
| dc.creator.none.fl_str_mv |
Costa, Lourena Emanuele Lima, Mayara Ingrid Sousa Chávez-Fumagalli, Miguel Angel Menezes-Souza, Daniel Martins, Vivian Tamietti Duarte, Mariana Costa Lage, Paula Sousa Lopes, Eliane Gonçalves Paiva Lage, Daniela P. Ribeiro, Tatiana Gomes Andrade, Pedro H R De Magalhães Soares, Danielle Ferreira Soto Álvarez, Manuel Tavares, Carlos Alberto Pereira Goulart, Luiz Ricardo Coelho, Eduardo Anton̂io Ferraz |
| author |
Costa, Lourena Emanuele |
| author_facet |
Costa, Lourena Emanuele Lima, Mayara Ingrid Sousa Chávez-Fumagalli, Miguel Angel Menezes-Souza, Daniel Martins, Vivian Tamietti Duarte, Mariana Costa Lage, Paula Sousa Lopes, Eliane Gonçalves Paiva Lage, Daniela P. Ribeiro, Tatiana Gomes Andrade, Pedro H R De Magalhães Soares, Danielle Ferreira Soto Álvarez, Manuel Tavares, Carlos Alberto Pereira Goulart, Luiz Ricardo Coelho, Eduardo Anton̂io Ferraz |
| author_role |
author |
| author2 |
Lima, Mayara Ingrid Sousa Chávez-Fumagalli, Miguel Angel Menezes-Souza, Daniel Martins, Vivian Tamietti Duarte, Mariana Costa Lage, Paula Sousa Lopes, Eliane Gonçalves Paiva Lage, Daniela P. Ribeiro, Tatiana Gomes Andrade, Pedro H R De Magalhães Soares, Danielle Ferreira Soto Álvarez, Manuel Tavares, Carlos Alberto Pereira Goulart, Luiz Ricardo Coelho, Eduardo Anton̂io Ferraz |
| author2_role |
author author author author author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Departamento de Biología Molecular Facultad de Ciencias Centro de Biología Molecular Severo Ochoa (CBM) |
| dc.subject.none.fl_str_mv |
Leishmaniasis Leishmania infantum Potential serodiagnosis applications Biología y Biomedicina / Biología |
| topic |
Leishmaniasis Leishmania infantum Potential serodiagnosis applications Biología y Biomedicina / Biología |
| description |
Visceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy and Trypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected with Leishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n 31) compared to those from vaccinated dogs (n 21), experimentally infected dogs with cross-reactive parasites (n 23), and healthy controls (n 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no crossreactivity with T. cruzi- or Ehrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes of L. infantum antigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programs |
| publishDate |
2014 |
| dc.date.none.fl_str_mv |
2014 2014-01-01 |
| dc.type.none.fl_str_mv |
research article http://purl.org/coar/resource_type/c_2df8fbb1 VoR http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10486/662771 https://dx.doi.org/10.1128/CVI.00583-13 |
| url |
http://hdl.handle.net/10486/662771 https://dx.doi.org/10.1128/CVI.00583-13 |
| dc.language.none.fl_str_mv |
Inglés eng |
| language_invalid_str_mv |
Inglés |
| language |
eng |
| dc.rights.none.fl_str_mv |
open access http://purl.org/coar/access_right/c_abf2 |
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info:eu-repo/semantics/openAccess |
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open access http://purl.org/coar/access_right/c_abf2 |
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openAccess |
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application/pdf |
| dc.publisher.none.fl_str_mv |
American Society for Microbiology. |
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American Society for Microbiology. |
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reponame:Biblos-e Archivo. Repositorio Institucional de la UAM instname:Universidad Autónoma de Madrid |
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Universidad Autónoma de Madrid |
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Biblos-e Archivo. Repositorio Institucional de la UAM |
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15.198674 |