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Nano-multilamellar lipid vesicles (NMVs) enhance protective antibody responses against Shiga toxin (Stx2a) produced by enterohemorrhagic Escherichia coli strains (EHEC)

  • M. J. Rodrigues-Jesus
  • W. L. Fotoran
  • R. M. Cardoso
  • K. Araki
  • G. Wunderlich
  • Luís C. S. Ferreira
Biotechnology and Industrial Microbiology - Research Paper

Abstract

Microlipid vesicles (MLV) have a broad spectrum of applications for the delivery of molecules, ranging from chemical compounds to proteins, in both in vitro and in vivo conditions. In the present study, we developed a new set of nanosize multilayer lipid vesicles (NMVs) containing a unique combination of lipids. The NMVs enable the adsorption of histidine-tagged proteins at the vesicle surface and were demonstrated to be suitable for the in vivo delivery of antigens. The NMVs contained a combination of neutral (DOPC) and anionic (DPPG) lipids in the inner membrane and an external layer composed of DOPC, cholesterol, and a nickel-containing lipid (DGS-NTA [Ni]). NMVs combined with a recombinant form of the B subunit of the Shiga toxin (rStx2B) produced by certain enterohemorragic Escherichia coli (EHEC) strains enhanced the immunogenicity of the antigen after parenteral administration to mice. Mice immunized with rStx2B-loaded NMVs elicited serum antibodies capable of neutralizing the toxic activities of the native toxin; this result was demonstrated both in vitro and in vivo. Taken together, these results demonstrated that the proposed NMVs represent an alternative for the delivery of antigens, including recombinant proteins, generated in different expression systems.

Keywords

Nanoparticles Delivery system Multilamellar vesicles Lipids vesicles Shiga toxin 

Notes

Acknowledgments

The authors would like to thank Eduardo Gimenez and Carolina Bertelli Ferreira for their technical support, Leticia Barbosa and Roxane Piazza of the Butantan Institute for the supply of the native Stx2a toxin used in this study, and Marina S. Palermo from the Faculty of Medicine at the University of Buenos Aires for supplying the pGEM-stx2 construction. All the data needed to evaluate the conclusions made in this paper are present within the data presented in the paper and/or the Supplemental Materials. Additional data may be requested from the authors.

Authors’ contributions

Jesus-Rodrigues, M.J. performed the rStx2B expression and purification assays, preparation of the NMVs, immunization and challenge of the mice, cell assays, antibody titers, data analysis, and wrote the manuscript. Fotoran, W.L. developed and assisted in the preparation of NMVs. Cardoso, R.M. carried out the physico-chemical characterization tests of NMVs. Araki, K. and Wunderlich, G. contributed scientific and technical assistance to the preparation and characterization of NMVs. Ferreira, L.C.S. supervised the study and the writing of the manuscript. All authors reviewed and commented on the manuscript.

Funding information

This work was carried out at the Institute of Biomedical Sciences of the University of São Paulo and with financial support of the Research Support Foundation of the States of São Paulo (FAPESP—Process: 2014/21141-4 and 2015/17174-7).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

42770_2018_35_MOESM1_ESM.docx (1.3 mb)
ESM 1 (DOCX 1.34 MB)

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Copyright information

© Sociedade Brasileira de Microbiologia 2018

Authors and Affiliations

  • M. J. Rodrigues-Jesus
    • 1
  • W. L. Fotoran
    • 2
  • R. M. Cardoso
    • 3
  • K. Araki
    • 3
  • G. Wunderlich
    • 2
  • Luís C. S. Ferreira
    • 1
  1. 1.Vaccine Development Laboratory, Department of MicrobiologyInstitute of Biomedical Sciences, University of São PauloSão PauloBrazil
  2. 2.Unit for Drug Development and Plasmodium Molecular Biology, Department of ParasitologyInstitute of Biomedical Sciences, University of São PauloSão PauloBrazil
  3. 3.Supramolecular Chemistry and Nanotechnology Laboratory, Department of Fundamental ChemistryInstitute of Chemistry, University of São PauloSão PauloBrazil

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