title: Mannosylated LigANI produced in Pichia pastoris protects hamsters against leptospirosis.
abstract: The C-terminal region of the Leptospiral immunoglobulin-like A protein (LigA) contains six carboxy-terminal Ig-like repeat domains (LigANI). Subunit vaccine preparations based on recombinant LigANI produced in Escherichia coli, are promising vaccine candidates, albeit with variable efficacy. In the present study, LigANI was expressed in the methylotrophic yeast Pichia pastoris using a 12 L bioreactor to produce mannosylated LigANI (mLigANI) for use in a vaccine preparation against leptospirosis. Hamsters immunized with a mLigANI vaccine preparation produced a significant IgG antibody response (P < 0.001) and were protected (83.3 %; P < 0.001) against lethal challenge with 36× LD50 of a virulent strain of L. interrogans serovar Copenhageni. A vaccine preparation based on demannosylated mLigANI (nmLigANI) elicited an immune response in hamsters, but did not afford protection. The production of mLigANI in bioreactor by P. pastoris yielded ~50 mg L(-1) of recombinant protein. P. pastoris is a potential platform for the production of leptospiral antigens on an industrial scale. The results demonstrate that LigANI secreted by P. pastoris on mannosylated form (mLigANI) protect hamsters as subunit vaccine of L. interrogans lethal infection.
authors.affiliation: Unidade de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil, daianehartwig@gmail.com.
authors.forename: Daiane D
authors.initials: DD
authors.lastname: Hartwig
authors.forename: Kátia L
authors.initials: KL
authors.lastname: Bacelo
authors.forename: Patrícia D de
authors.initials: PD
authors.lastname: Oliveira
authors.forename: Thaís L
authors.initials: TL
authors.lastname: Oliveira
authors.forename: Fabiana K
authors.initials: FK
authors.lastname: Seixas
authors.forename: Marta G
authors.initials: MG
authors.lastname: Amaral
authors.forename: Cláudia P
authors.initials: CP
authors.lastname: Hartleben
authors.forename: Alan J A
authors.initials: AJ
authors.lastname: McBride
authors.forename: Odir A
authors.initials: OA
authors.lastname: Dellagostin
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019149:Bioreactors; D006224:Cricetinae; D005260:Female; D007919:Leptospira / Q000737:chemistry* / Q000235:genetics; D007922:Leptospirosis / Q000276:immunology / Q000517:prevention & control*; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016019:Survival Analysis
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins