title: Innovative immunization protocols using chimeric recombinant protein for the production of polyspecific loxoscelic antivenom in horses.
abstract: A chimeric protein (rCpLi) was constructed expressing three epitopes of rLiD1, a dermonecrotic toxin from the venom of Loxosceles intermedia spider. We have analyzed the neutralization potential of sera obtained by immunization of horses with rCpLi and rCpLi combined with initial doses of venoms and compared these with antivenom traditionally produced in horses using crude Loxosceles gaucho, Loxosceles laeta and L. intermedia venoms as antigens. We have demonstrated by ELISA that horses immunized with three initial doses of crude venom containing mixtures of L. intermedia, L. gaucho and L. laeta followed by nine doses of rCpLi generate antibodies with the same reactivity as those produced following immunization with traditional antivenom, towards the venoms of the three Loxosceles sp. species. Results from in vivo and in vitro neutralization assays showed that the new horse sera are able to neutralize the dermonecrotic activity of Loxosceles venoms, which are of medical importance in Brazil and some of these sera are capable of meeting the necessary potency requirements that could allow for their therapeutic use in humans. This immunization strategy combining both antigens used approximately 67% less crude Loxosceles venoms compared to traditional immunization protocol and can mean the development of Loxosceles antivenoms with the consequent reduction of devastation of arachnid fauna.
authors.affiliation: Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil; Centro de Produção e Pesquisa de Imunobiológicos (CPPI), Piraquara, PR, Brazil.
authors.forename: Luís F M
authors.initials: LF
authors.lastname: Figueiredo
authors.affiliation: Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: Camila
authors.initials: C
authors.lastname: Dias-Lopes
authors.affiliation: Universidade Federal do Paraná - Setor de Ciências Biológicas, Departamento de Patologia Básica, Curitiba, PR, Brazil.
authors.forename: Larissa M
authors.initials: LM
authors.lastname: Alvarenga
authors.affiliation: Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: Thais M
authors.initials: TM
authors.lastname: Mendes
authors.affiliation: Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: Ricardo A
authors.initials: RA
authors.lastname: Machado-de-Ávila
authors.affiliation: National Heart and Lung Institute, Faculty of Medicine, Imperial College London, Sir Alexander Fleming Building, London SW7 2AZ, UK.
authors.forename: Jessica
authors.initials: J
authors.lastname: McCormack
authors.affiliation: Centro de Produção e Pesquisa de Imunobiológicos (CPPI), Piraquara, PR, Brazil.
authors.forename: João C
authors.initials: JC
authors.lastname: Minozzo
authors.affiliation: Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil.
authors.forename: Evanguedes
authors.initials: E
authors.lastname: Kalapothakis
authors.affiliation: Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP 486, CEP 31270-901, Belo Horizonte, MG, Brazil. Electronic address: olortegi@icb.ufmg.br.
authors.forename: Carlos
authors.initials: C
authors.lastname: Chávez-Olórtegui
mesh_terms: D000818:Animals; D000997:Antivenins / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D006736:Horses / Q000276:immunology*; D007114:Immunization / Q000379:methods / Q000662:veterinary; D009500:Neutralization Tests; D010727:Phosphoric Diester Hydrolases / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D013111:Spider Venoms / Q000276:immunology*
chemical_list: D000997:Antivenins; D011993:Recombinant Fusion Proteins; D013111:Spider Venoms; C055161:loxosceles venom; D010727:Phosphoric Diester Hydrolases
keywords: B-cell epitopes; Loxoscelic antivenom; Recombinant chimeric protein; Sphingomyelinase D