Subtractive Phage Display Selection from Canine Visceral Leishmaniasis Identifies Novel Epitopes That Mimic Leishmania infantum Antigens with Potential Serodiagnosis Applications

Subtractive Phage Display Selection from Canine Visceral Leishmaniasis Identifies Novel Epitopes That Mimic Leishmania infantum Antigens with Potential Serodiagnosis Applications

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Published in:Clinical and Vaccine Immunology Vol. 21; no. 1; pp. 96 - 106
Main Authors: Costa, Lourena E, Lima, Mayara I S, Chávez-Fumagalli, Miguel A, Menezes-Souza, Daniel, Martins, Vivian T, Duarte, Mariana C, Lage, Paula S, Lopes, Eliane G P, Lage, Daniela P, Ribeiro, Tatiana G, Andrade, Pedro H R, de Magalhães-Soares, Danielle F, Soto, Manuel, Tavares, Carlos A P, Goulart, Luiz R, Coelho, Eduardo A F
Format: Journal Article
Language:English
Published: United States American Society for Microbiology 01-01-2014
Subjects:
Animals
Antibodies, Protozoan - blood
Antigens, Protozoan - isolation & purification
Brazil
Diagnostic Laboratory Immunology
Dog Diseases - diagnosis
Dog Diseases - parasitology
Dogs
Ehrlichia
Enzyme-Linked Immunosorbent Assay - methods
Epitopes - isolation & purification
Female
Leishmania infantum
Leishmania infantum - immunology
Leishmaniasis, Visceral - diagnosis
Leishmaniasis, Visceral - parasitology
Leishmaniasis, Visceral - veterinary
Male
Peptide Library
Peptides - isolation & purification
Predictive Value of Tests
Sensitivity and Specificity
Serologic Tests - methods
Trypanosoma
Brazil
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Abstract Classifications Services CVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue CVI About CVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy CVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 1556-6811 Online ISSN: 1556-679X Copyright © 2014 by the American Society for Microbiology.   For an alternate route to CVI .asm.org, visit: CVI       
AbstractList 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 cross-reactivity 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.
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 cross-reactivity 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.
Classifications Services CVI Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue CVI About CVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy CVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 1556-6811 Online ISSN: 1556-679X Copyright © 2014 by the American Society for Microbiology.   For an alternate route to CVI .asm.org, visit: CVI       
Author Daniel Menezes-Souza
Vivian T. Martins
Pedro H. R. Andrade
Eliane G. P. Lopes
Danielle F. de Magalhães-Soares
Carlos A. P. Tavares
Daniela P. Lage
Miguel A. Chávez-Fumagalli
Manuel Soto
Eduardo A. F. Coelho
Paula S. Lage
Lourena E. Costa
Mayara I. S. Lima
Mariana C. Duarte
Luiz R. Goulart
Tatiana G. Ribeiro
Author_xml – sequence: 1
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  surname: Costa
  fullname: Costa, Lourena E
  organization: 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
– sequence: 2
  givenname: Mayara I S
  surname: Lima
  fullname: Lima, Mayara I S
– sequence: 3
  givenname: Miguel A
  surname: Chávez-Fumagalli
  fullname: Chávez-Fumagalli, Miguel A
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  surname: Martins
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  fullname: Andrade, Pedro H R
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  surname: de Magalhães-Soares
  fullname: de Magalhães-Soares, Danielle F
– sequence: 13
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  fullname: Goulart, Luiz R
– sequence: 16
  givenname: Eduardo A F
  surname: Coelho
  fullname: Coelho, Eduardo A F
BackLink https://www.ncbi.nlm.nih.gov/pubmed/24256622$$D View this record in MEDLINE/PubMed
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Visceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of...
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StartPage 96
SubjectTerms Animals
Antibodies, Protozoan - blood
Antigens, Protozoan - isolation & purification
Brazil
Diagnostic Laboratory Immunology
Dog Diseases - diagnosis
Dog Diseases - parasitology
Dogs
Ehrlichia
Enzyme-Linked Immunosorbent Assay - methods
Epitopes - isolation & purification
Female
Leishmania infantum
Leishmania infantum - immunology
Leishmaniasis, Visceral - diagnosis
Leishmaniasis, Visceral - parasitology
Leishmaniasis, Visceral - veterinary
Male
Peptide Library
Peptides - isolation & purification
Predictive Value of Tests
Sensitivity and Specificity
Serologic Tests - methods
Trypanosoma
Title Subtractive Phage Display Selection from Canine Visceral Leishmaniasis Identifies Novel Epitopes That Mimic Leishmania infantum Antigens with Potential Serodiagnosis Applications
URI http://cvi.asm.org/content/21/1/96.abstract
https://www.ncbi.nlm.nih.gov/pubmed/24256622
https://search.proquest.com/docview/1490777542
https://search.proquest.com/docview/1492626268
https://pubmed.ncbi.nlm.nih.gov/PMC3910914
Volume 21
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