title: Linear B-cell epitopes in BthTX-1, BthTX-II and BthA-1, phospholipase A₂'s from Bothrops jararacussu snake venom, recognized by therapeutically neutralizing commercial horse antivenom.
abstract: The benefits from treatment with antivenom sera are indubitable. However, the mechanism for toxin neutralization has not been completely elucidated. A mixture of anti-bothropic and anti-crotalic horse antivenom has been reported to be more effective in neutralizing the effects of Bothrops jararacussu snake venom than anti-bothropic antivenom alone. This study determined which regions in the three PLA₂s from B. jararacussu snake venom are bound by antibodies in tetravalent anti-bothropic and monovalent anti-crotalic commercial horse antivenom. Mapping experiments of BthTX-I, BthTX-II and BthA-I using two small libraries of 69 peptides each revealed six major IgG-binding epitopes that were recognized by both anti-bothropic and anti-crotalic horse antivenom. Two epitopes in BthTX-I were only recognized by the anti-bothropic horse antivenom, while anti-crotalic horse antivenom recognized four unique epitopes across the three PLA₂s. Our studies suggest that the harmful activities of the PLA₂s present in the venom of B. jararacussu are neutralized by the combinatorial treatment with both antivenom sera through their complementary binding sites, which provides a wide coverage on the PLA₂s. This is the first peptide microarray of PLA₂s from B. jararacussu snake venom to survey the performance of commercial horse antiophidic antivenom. Regions recognized by the protective antivenom sera are prime candidates for improved venom cocktails or a chimeric protein encoding the multiple epitopes to immunize animals as well as for designing future synthetic vaccines.
authors.affiliation: Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças Negligenciadas-INCT-IDN, Centro de Desenvolvimento Tecnológico em Saúde-CDTS, FIOCRUZ, Rio de Janeiro, RJ, Brazil. dsimone@ioc.fiocruz.br
authors.forename: Salvatore G
authors.initials: SG
authors.lastname: De-Simone
authors.forename: Paloma
authors.initials: P
authors.lastname: Napoleão-Pego
authors.forename: Luiz A L
authors.initials: LA
authors.lastname: Teixeira-Pinto
authors.forename: Jonathas D L
authors.initials: JD
authors.lastname: Santos
authors.forename: Thatiane S
authors.initials: TS
authors.lastname: De-Simone
authors.forename: Anibal R
authors.initials: AR
authors.lastname: Melgarejo
authors.forename: Aniesse S
authors.initials: AS
authors.lastname: Aguiar
authors.forename: Daniela P
authors.initials: DP
authors.lastname: Marchi-Salvador
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000997:Antivenins / Q000737:chemistry; D017837:Bothrops; D003429:Cross Reactions; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D054501:Group II Phospholipases A2 / Q000737:chemistry / Q000276:immunology*; D006736:Horses / Q000276:immunology*; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D019151:Peptide Library; D054467:Phospholipases A2 / Q000737:chemistry / Q000276:immunology*; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D000997:Antivenins; D003435:Crotalid Venoms; D018985:Epitopes, B-Lymphocyte; D019151:Peptide Library; C078097:bothropstoxin; D054501:Group II Phospholipases A2; D054467:Phospholipases A2; C072031:bothropstoxin II
keywords: Antigenic determinants; B-epitopes; Peptide array; Phospholipase A(2); SPOT-synthesis; Synthetic peptides