Lactococcus lactis IBB477 presenting adhesive and muco-adhesive properties as a candidate carrier strain for oral vaccination against influenza virus.

Abstract

In the gastrointestinal tract (GIT), adhesion is a prerequisite for bacterial colonization. Lactococci can be used in functional food (probiotics) and health-related applications (mucosal vaccines, therapeutic drug delivery), both potentially involving adhesive properties. A candidate lactic acid bacterium for influenza antigen delivery through the GIT should display the ability to adhere. The present work probes the interactions between Lactococcus lactis and mucins using pig gastric mucin (PGM) as a model. Two strains were used for the optimization of the screening method for adhesion: L. lactis subsp. cremoris IBB477 persistent in the GIT of germ-free rats, and the low-adhering control strain MG1820. High adhesion to bare and mucin-coated polystyrene of IBB477 in comparison with MG1820 was observed. We searched for genetic determinants potentially involved in the adhesion/ muco-adhesion of IBB477, identifying two such genes: prtP and a gene coding for a protein with MUB and MucBP domains. Based on its persistence in the GIT and adhesive properties, L. lactis IBB477 is a candidate carrier strain for expression of influenza haemagglutinin (HA) protein for induction of mucosal immune response.

In vitro characteristics of Lactobacillus spp. strains isolated from the chicken digestive tract and their role in the inhibition of Campylobacter colonization

“…Campylobacter growth inhibition by supernatant of Lactobacillus strains is marked as +. Bacterial adhesion was determined using the technique described by Radziwill-Bienkowska et al (2014) in three independent experiments. Bacteria were characterized as strongly adherent (A ≥ 6), moderately adherent (6 > A ≥ 3), weakly adherent (3 > A > 2), and nonadherent (A ≤ 2).…”

“…Adhesion of bacterial cells to polystyrene (PS) was tested on the 96‐well microtiter PS plates (Thermo Fischer Scientific Nunc A/S, Denmark) using a modification of the method already described (Radziwill‐Bienkowska, Zochowska, Bardowski, Mercier‐Bonin, & Kowalczyk, ). Briefly, bacteria from overnight cultures diluted to OD 660 nm of 1.0 were harvested by centrifugation (10 3 g ; 1 min) and resuspended in an equal volume of PBS (BioShop, Canada).…”

“…It is postulated that highly adhesive probiotic bacteria have the greatest beneficial effects on host health and they should, at least transiently, colonize the host gut (Bermudez-Brito et al, 2012). The adhesion microtiter plate assay is a well-known and simple method for a preliminary assessment of strain adhesion ability (Radziwill-Bienkowska et al, 2014), which, as we observed in our previous studies, can correlate with its adhesiveness to biotic surfaces Radziwill-Bienkowska et al, 2014. In this study, we checked the adhesive properties of 64 Lactobacillus strains isolated from chicken gastrointestinal tracts and compared them with highly adhesive or nonadhesive L. rhamnosus strains (GG or LOCK 0908, respectively).…”

“…Radziwill-Bienkowska, Zochowska, Bardowski, Mercier-Bonin, & Kowalczyk, 2014). Briefly, bacteria from overnight cultures diluted to OD 660 nm of 1.0 were harvested by centrifugation (10 3 g; 1 min) and re-suspended in an equal volume of PBS (BioShop, Canada).…”

Potential of Lactobacillus plantarum IBB3036 and Lactobacillus salivarius IBB3154 to persistence in chicken after in ovo delivery

“…Bacteria were evaluated for their adhesion ability to bare polystyrene in 96‐well microtiter plates (Thermo Fischer Scientific Nunc A/S, Denmark) using the technique described by (Christensen, Baldassarri, & Simpson, ) and modified by Radziwill‐Bienkowska (Radziwill‐Bienkowska et al. ). Each strain was tested in three independent experiments, and the results are presented as the means ± standard deviations.…”