title: Generation and characterization of a recombinant chimeric protein (rCpLi) consisting of B-cell epitopes of a dermonecrotic protein from Loxosceles intermedia spider venom.
abstract: A chimeric protein was constructed expressing three epitopes of LiD1, a dermonecrotic toxin from the venom of Loxosceles intermedia spider. This species is responsible for a large number of accidents involving spiders in Brazil. We demonstrated that the chimeric protein (rCpLi) generated is atoxic and that antibodies previously developed in rabbits against synthetic epitopes reactive with rCpLi in ELISA and immunoblot assays. The antibody response in rabbits against the rCpLi was evaluated by ELISA and we have detected an antibody response in all immunized animals. Overlapping peptides covering the amino acid sequence of the rCpLi were synthesized on a cellulose membrane, and their recognition by rabbit anti-rCpLi serum assessed. Three different antigenic regions were identified. The percentage of inhibition of the dermonecrotic, hemorrhagic and edematogenic activities caused by the recombinant protein LiD1r in naïve rabbits was assessed by pre-incubation with anti-rCpLi antibodies. Anti-rCpLi induced good dermonecrotic and hemorrhagic protection. The levels of protection were similar to the antiboides anti-LiD1r. In summary, we have developed a polyepitope recombinant chimeric protein capable of inducing multiple responses of neutralizing antibodies in a rabbit model. This engineered protein may be a promising candidate for therapeutic serum development or vaccination.
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: T M
authors.initials: TM
authors.lastname: Mendes
authors.forename: D
authors.initials: D
authors.lastname: Oliveira
authors.forename: L F M
authors.initials: LF
authors.lastname: Figueiredo
authors.forename: R A
authors.initials: RA
authors.lastname: Machado-de-Avila
authors.forename: C G
authors.initials: CG
authors.lastname: Duarte
authors.forename: C
authors.initials: C
authors.lastname: Dias-Lopes
authors.forename: G
authors.initials: G
authors.lastname: Guimarães
authors.forename: L
authors.initials: L
authors.lastname: Felicori
authors.forename: J C
authors.initials: JC
authors.lastname: Minozzo
authors.forename: C
authors.initials: C
authors.lastname: Chávez-Olortegui
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D000997:Antivenins / Q000276:immunology; D004487:Edema / Q000276:immunology / Q000517:prevention & control; D018985:Epitopes, B-Lymphocyte / Q000235:genetics; D006470:Hemorrhage / Q000276:immunology / Q000517:prevention & control; D009336:Necrosis / Q000276:immunology / Q000517:prevention & control; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D012867:Skin / Q000276:immunology / Q000473:pathology; D013108:Sphingomyelin Phosphodiesterase / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D013111:Spider Venoms / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D013112:Spiders
chemical_list: D057134:Antibodies, Neutralizing; D000997:Antivenins; D018985:Epitopes, B-Lymphocyte; D011993:Recombinant Fusion Proteins; D013111:Spider Venoms; C516323:dermonecrotic toxin, Loxosceles intermedia; D013108:Sphingomyelin Phosphodiesterase