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Mannosylated LigANI Produced in Pichia pastoris Protects Hamsters Against Leptospirosis

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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.

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Acknowledgments

We are grateful to Michele dos Santos and Kátia R. Pimenta Cardoso for technical assistance.

This work was supported by CNPq (Grant Number 475830/2012-1) and FAPERGS/ARD-no 003/2012. AJAM and OAD were recipients of research scholarships from CNPq.

Conflict of interest

AJAM and OAD are inventors on a patent submission entitled: LigA and LigB proteins (Leptospiral Ig-like (Lig) domains) for vaccination and diagnosis (Patent nos. BRPI0505529 and WO 2007070996). The other authors declare no conflict of interest.

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Authors and Affiliations

  1. Unidade de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil

    Daiane D. Hartwig, Kátia L. Bacelo, Patrícia D. de Oliveira, Thaís L. Oliveira, Fabiana K. Seixas, Marta G. Amaral, Cláudia P. Hartleben, Alan J. A. McBride & Odir A. Dellagostin

  2. Departamento de Microbiologia e Parasitologia, Instituto de Biologia, Universidade Federal de Pelotas, Campus Universitário, Pelotas, RS, 96010-900, Brazil

    Daiane D. Hartwig

  3. Instituto Gonçalo Moniz, Fiocruz-BA, Ministério da Saúde, Salvador, BA, Brazil

    Alan J. A. McBride

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  1. Daiane D. Hartwig
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  2. Kátia L. Bacelo
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  3. Patrícia D. de Oliveira
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Correspondence to Daiane D. Hartwig.

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Hartwig, D.D., Bacelo, K.L., de Oliveira, P.D. et al. Mannosylated LigANI Produced in Pichia pastoris Protects Hamsters Against Leptospirosis. Curr Microbiol 68, 524–530 (2014). https://doi.org/10.1007/s00284-013-0505-4

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  • DOI: https://doi.org/10.1007/s00284-013-0505-4

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