Rotavirus VP6 protein mucosally delivered by cell wall-derived particles from Lactococcus lactis induces protection against infection in a murine model

PLoS One. 2018 Sep 7;13(9):e0203700. doi: 10.1371/journal.pone.0203700. eCollection 2018.

Abstract

Rotaviruses are the primary cause of acute gastroenteritis in children worldwide. Although the implementation of live attenuated vaccines has reduced the number of rotavirus-associated deaths, variance in their effectiveness has been reported in different countries. This fact, among other concerns, leads to continuous efforts for the development of new generation of vaccines against rotavirus.In this work, we describe the obtention of cell wall-derived particles from a recombinant Lactococcus lactis expressing a cell wall-anchored version of the rotavirus VP6 protein. After confirming by SDS-PAGE, Western blot, flow cytometry and electronic immunomicroscopy that these particles were carrying the VP6 protein, their immunogenic potential was evaluated in adult BALB/c mice. For that, mucosal immunizations (oral or intranasal), with or without the dmLT [(double mutant Escherichia coli heat labile toxin LT(R192G/L211A)] adjuvant were performed. The results showed that these cell wall-derived particles were able to generate anti-rotavirus IgG and IgA antibodies only when administered intranasally, whether the adjuvant was present or not. However, the presence of dmLT was necessary to confer protection against rotavirus infection, which was evidenced by a 79.5 percent viral shedding reduction.In summary, this work describes the production of cell wall-derived particles which were able to induce a protective immune response after intranasal immunization. Further studies are needed to characterize the immune response elicited by these particles as well as to determine their potential as an alternative to the use of live L. lactis for mucosal antigen delivery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity
  • Antigens, Viral / immunology*
  • Antigens, Viral / metabolism
  • Capsid Proteins / immunology*
  • Capsid Proteins / metabolism
  • Cell Wall / metabolism*
  • Disease Models, Animal
  • Drug Carriers / metabolism*
  • Immunoglobulin A / immunology
  • Immunoglobulin G / immunology
  • Lactococcus lactis / cytology*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mucous Membrane / metabolism*
  • Rotavirus / physiology*
  • Rotavirus Infections / prevention & control*

Substances

  • Antigens, Viral
  • Capsid Proteins
  • Drug Carriers
  • Immunoglobulin A
  • Immunoglobulin G
  • VP6 protein, Rotavirus

Grants and funding

This study was supported by PICT-2012-300 to C.F.T from Agencia Nacional de Promoción Científica y Tecnológica (ANCyPT), Argentina; and grant from the Universidad Nacional de Quilmes for the research program “Basic and Applied Molecular Microbiology” (PUNQ1407/15) and the research grants issued by Universidad Nacional Arturo Jauretche, Res.(R) 290/12 and 404/15. Doctoral and Postdoctoral Fellowship from CONICET for D. Silvestre and M.G. Mandile, respectively, are also acknowledged. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.