Alexandre Antunes Brum1, Andrea de Fatima Silva Rezende1, Francisco Silvestre Brilhante1, Thais Collares1, Karine Begnine2, Fabiana Kommling Seixas2, Tiago Veiras Collares2, Odir Antônio Dellagostin3, Vasco Azevedo4, Anderson Santos5, Ricardo Wagner Portela6, Sibele Borsuk1. 1. Laboratório de Biotecnologia Infecto-parasitária, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil. 2. Grupo de Pesquisa em Oncologia Celular e Molecular, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil. 3. Laboratório de Biologia Molecular, Centro de Desenvolvimento Tecnológico, Biotecnologia, UFPel, Pelotas, RS 96010-900, Brazil. 4. Laboratório de Genética Celular e Molecular, Instituto de Ciências Biológicas, UFMG, Belo Horizonte, MG 31270-901, Brazil. 5. Faculdade de Computação, UFU, Uberlândia, MG 38400-902, Brazil. 6. Laboratório de Imunologia e Biologia Molecular, Instituto de Ciências da Saúde, UFBA, Salvador, BA 40110-100, Brazil.
Abstract
PURPOSE: We tested the efficacy of the esterase encoded by cp1002_RS09720 from Corynebacteriumpseudotuberculosis in recombinant subunit and DNA caseous lymphadenitis (CLA) vaccines. This target was predicted as one of the best CLA vaccine candidates by mature epitope density analysis. METHODOLOGY: Gene cp1002_RS09720 was cloned into two different vectors (pAE for subunit vaccine and pTARGET for DNA vaccine). Four groups of 15 mice each were immunized with the recombinant esterase rCP09720 associated with aluminium hydroxide adjuvant (G1), pTARGET/cp09720 DNA vaccine (G2), a naked pTARGET (G3) or PBS as a negative control (G4). Immunization occurred in two doses intercalated by a 21 day interval. Twenty-one days after the last dose administration, animals were challenged with a virulent C. pseudotuberculosis MIC-6 strain. RESULTS: G1 showed high levels of IgG1 and IgG2a on days 21 and 42 post-immunization and a significant level of IFN-γ (P<0.05), suggesting a Th1 response. The protection levels obtained were 58.3 and 16.6 % for G1 and G2, respectively. CONCLUSION: The subunit vaccine composed of the recombinant esterase rCP09720 and Al(OH)3 is a promising antigenic formulation for use against CLA.
PURPOSE: We tested the efficacy of the esterase encoded by cp1002_RS09720 from Corynebacteriumpseudotuberculosis in recombinant subunit and DNA caseous lymphadenitis (CLA) vaccines. This target was predicted as one of the best CLA vaccine candidates by mature epitope density analysis. METHODOLOGY: Gene cp1002_RS09720 was cloned into two different vectors (pAE for subunit vaccine and pTARGET for DNA vaccine). Four groups of 15 mice each were immunized with the recombinant esterase rCP09720 associated with aluminium hydroxide adjuvant (G1), pTARGET/cp09720 DNA vaccine (G2), a naked pTARGET (G3) or PBS as a negative control (G4). Immunization occurred in two doses intercalated by a 21 day interval. Twenty-one days after the last dose administration, animals were challenged with a virulent C. pseudotuberculosis MIC-6 strain. RESULTS: G1 showed high levels of IgG1 and IgG2a on days 21 and 42 post-immunization and a significant level of IFN-γ (P<0.05), suggesting a Th1 response. The protection levels obtained were 58.3 and 16.6 % for G1 and G2, respectively. CONCLUSION: The subunit vaccine composed of the recombinant esterase rCP09720 and Al(OH)3 is a promising antigenic formulation for use against CLA.
Authors: Alberto Oliveira; Leticia C Oliveira; Flavia Aburjaile; Leandro Benevides; Sandeep Tiwari; Syed B Jamal; Arthur Silva; Henrique C P Figueiredo; Preetam Ghosh; Ricardo W Portela; Vasco A De Carvalho Azevedo; Alice R Wattam Journal: Front Microbiol Date: 2017-10-12 Impact factor: 5.640