title: Use of Phage Display technology in development of canine visceral leishmaniasis vaccine using synthetic peptide trapped in sphingomyelin/cholesterol liposomes.
abstract: BACKGROUND
  Leishmania parasites can cause visceral or cutaneous disease and are found in subtropical and tropical regions of the Old and New World. The pathology of the infection is determined by both host immune factors and species/strain differences of the parasite. Dogs represent the major reservoir of Leishmania infantum (syn. L. chagasi) and vaccines are considered the most cost-effective control tools for canine disease.
  
  
  METHODS
  Selection of immunodominant peptides was performed by Phage Display to identify sequences recognized by L. infantum naturally infected animals. Sera from Leishmania infected animals were used in the biopanning to selection of specific peptides. Serum samples from T. cruzi infected and healthy animals were used as control. After selection, synthetic peptides were produced in membrane (spot-synthesis) in soluble form and blotting and ELISA were performed for validation of serum reactivity. Selected peptide was formulated with aluminum hydroxide and liposomes and immunization was performed in BALB/c mice. Protection was determined by qPCR after challenge infection with virulent L. infantum.
  
  
  RESULTS
  We reported the selection of Peptide 5 through Phage Display technique and demonstrate its ability to promote a state of immunity against L. infantum infection in murine model after immunization using liposomes as vaccine carrier. Our results demonstrate that immunization with Peptide 5 when formulated with aluminum hydroxide and liposomes is immunogenic and elicited significant protection associated with the induction of mixed Th1/Th2 immune response against L. infantum infection.
  
  
  CONCLUSIONS
  Peptide 5 is a promising vaccine candidate and the findings obtained in the present study encourage canine trials to confirm the effectiveness of a vaccine against CVL.
authors.affiliation: Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. cmonerat@yahoo.com.br.
authors.forename: Christina Monerat
authors.initials: CM
authors.lastname: Toledo-Machado
authors.affiliation: Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. lilacerdabueno@gmail.com.
authors.forename: Lilian Lacerda
authors.initials: LL
authors.lastname: Bueno
authors.affiliation: Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. daniel.ufop@gmail.com.
authors.forename: Daniel
authors.initials: D
authors.lastname: Menezes-Souza
authors.affiliation: Unidade Acadêmica de Ciências da Saúde, Universidade do Extremo Sul Catarinense - CEP: 88.806-000, Criciúma, Santa Catarina, Brazil. randrez@yahoo.com.br.
authors.forename: Ricardo Andrez
authors.initials: RA
authors.lastname: Machado-de-Avila
authors.affiliation: SysDiag CNRS-BioRad UMR 3145, Cap Delta/Parc Euromédecine, 1682 rue de la Valsière, CS 61003, 34184, Montpellier Cedex 4, France. nguyen@sysdiag.cnrs.fr.
authors.forename: Christophe
authors.initials: C
authors.lastname: Nguyen
authors.affiliation: SysDiag CNRS-BioRad UMR 3145, Cap Delta/Parc Euromédecine, 1682 rue de la Valsière, CS 61003, 34184, Montpellier Cedex 4, France. claude_granier@bbox.fr.
authors.forename: Claude
authors.initials: C
authors.lastname: Granier
authors.affiliation: Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. daniella@icb.ufmg.br.
authors.forename: Daniella Castanheira
authors.initials: DC
authors.lastname: Bartholomeu
authors.affiliation: Departamento Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. olortegi@icb.ufmg.br.
authors.forename: Carlos
authors.initials: C
authors.lastname: Chávez-Olórtegui
authors.affiliation: Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. fujiwara@icb.ufmg.br.
authors.forename: Ricardo Toshio
authors.initials: RT
authors.lastname: Fujiwara
mesh_terms: D000818:Animals; D002784:Cholesterol; D004283:Dog Diseases / Q000276:immunology* / Q000469:parasitology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D018314:Leishmania infantum / Q000276:immunology*; D054332:Leishmaniasis Vaccines / Q000276:immunology*; D007898:Leishmaniasis, Visceral / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*; D008081:Liposomes; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D019151:Peptide Library; D010455:Peptides / Q000276:immunology*; D013109:Sphingomyelins
chemical_list: D054332:Leishmaniasis Vaccines; D008081:Liposomes; D019151:Peptide Library; D010455:Peptides; D013109:Sphingomyelins; D002784:Cholesterol