Inhibition of Streptococcus pneumoniae adherence to human epithelial cells in vitro by the probiotic Lactobacillus rhamnosus GG

Wong, Sook‐San; Toh, Zheng Quan; Dunne, Eileen M.; Mulholland, E. Kim; Tang, Mimi L.K.; Robins-Browne, Roy M.; Licciardi, Paul V.; Satzke, Catherine

doi:10.1186/1756-0500-6-135

Cited by 37 publications

(32 citation statements)

References 46 publications

(41 reference statements)

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“…As described here, the assay does not distinguish between adherent and invasive (internalized) pneumococci. However, previous studies have found the levels of internalized pneumococci to be very small (typically ≤0.01% of inocula) 10 so for most isolates (including PMP843), the vast majority of cell-associated pneumococci are adherent rather than internalized. Additional applications include testing other probiotic species for the ability to inhibit colonization, and/or looking at mutant bacterial strains to investigate the role of specific genes of interest.…”

Section: Discussionmentioning

confidence: 99%

“…In vitro adherence assays have been used to examine pneumococcal adherence 8,9 . These assays have been adapted to investigate the impact of the probiotic Lactobacillus rhamnosus GG (LGG) on pneumococcal adherence 10 .…”

Section: Introductionmentioning

confidence: 99%

“…The protocol presented here describes an adherence assay used to investigate the effects of S. salivarius K12 on pneumococcal adherence to the human epithelial cell line CCL-23. Prior to use in the assay, pneumococcal isolates were evaluated for adherence capabilities, as growth and adherence properties can vary substantially among isolates 10,14 . Growth curves of pneumococci and S. salivarius were performed to determine mid-log phase and estimate concentration (colony forming units or CFU/ml) by optical density (OD, Figure 1).…”

Section: Introductionmentioning

confidence: 99%

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Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Dunne

Toh

John

et al. 2014

JoVE

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Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a prerequisite for pneumococcal infections such as pneumonia and otitis media. In vitro adherence assays can be used to study the attachment of pneumococci to epithelial cell monolayers and to investigate potential interventions, such as the use of probiotics, to inhibit pneumococcal colonization. The protocol described here is used to investigate the effects of the probiotic Streptococcus salivarius on the adherence of pneumococci to the human epithelial cell line CCL-23 (sometimes referred to as HEp-2 cells). The assay involves three main steps: 1) preparation of epithelial and bacterial cells, 2) addition of bacteria to epithelial cell monolayers, and 3) detection of adherent pneumococci by viable counts (serial dilution and plating) or quantitative real-time PCR (qPCR). This technique is relatively straightforward and does not require specialized equipment other than a tissue culture setup. The assay can be used to test other probiotic species and/or potential inhibitors of pneumococcal colonization and can be easily modified to address other scientific questions regarding pneumococcal adherence and invasion. Video LinkThe video component of this article can be found at

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Dunne

Toh

John

et al. 2014

JoVE

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“…These mechanisms lead to the modulation of the microbiota and/or inhibition of colonization by potential pathogens, and bacterial translocation across the intestinal wall (Eizaguirre et al, 2011;Wine et al, 2009;Wong et al, 2013).…”

Section: Probioticsmentioning

confidence: 99%

Labneh with probiotic properties produced from kefir: development and sensory evaluation

Rocha¹,

Martins²,

Santos³

et al. 2014

Food Sci. Technol (Campinas)

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The labneh or labaneh is a popular fermented milk in the Middle East. Another fermented product that deserves special mention is kefir since it has probiotic activity and unique sensory, nutritional, and therapeutic properties. The aim of the present study was to develop a functional probiotic labneh using kefir as a fermenting agent and to perform a sensory analysis of the obtained product. Kefir was obtained by growing grains in pasteurized milk. Samples of skimmed and whole labneh were prepared from the inoculation of 5% kefir milk (skimmed/whole) at 28 °C for 24h, followed by cooling (12-18h) and whey drainage (12-24h), both at 4 °C. Sensory analysis was performed with 70 untrained panelists using a 9-point scale hedonic in the acceptance tests. The paired t-test was used to compare the differences between the means of the scores obtained, with the significance level of 5%. The labneh prepared showed good acceptance by the judges, and the whole labneh samples had the highest scores in the acceptance test. Further studies on the analysis of microbiological viability, nutritional composition, and determination of shelf life, also to improve acceptability of the low-fat version of the product, are needed.

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“…Recent works have investigated the effect of probiotics on pathogen-mediated cytotoxicity in mammalian cells using the CFS approach (Wong et al, 2013;Bermudez-Brito et al, 2014). As the impact of an individual probiotic is influenced by the composition of the microbiota (Howarth & Wang, 2013), the present investigation included both CFS and CFSC.…”

Section: Cytotoxicity Of E Coli O157 : H7mentioning

confidence: 99%

Lactobacillus plantarum LB95 impairs the virulence potential of Gram-positive and Gram-negative food-borne pathogens in HT-29 and Vero cell cultures

Dutra

Silva

Cabrita

et al. 2016

Journal of Medical Microbiology

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Listeria monocytogenes, Salmonella enterica and verocytotoxigenic Escherichia coli (VTEC) are amongst the most important agents responsible for food outbreaks occurring worldwide. In this work, two Lactobacillus spp. strains (LABs), Lactobacillus plantarum (LB95) and Lactobacillus paraplantarum (LB13), previously isolated from spontaneously fermenting olive brines, and two reference probiotic strains, Lactobacillus casei Shirota and Lactobacillus rhamnosus GG, were investigated for their ability to attenuate the virulence of the aforementioned pathogens using animal cell culture assays. In competitive exclusion assays, the relative percentages of adhesion and invasion of S. enterica subsp. enterica serovar Enteritidis were significantly reduced when the human HT-29 cell line was previously exposed to LB95. The relative percentage of invasion by Listeria monocytogenes was significantly reduced when HT-29 cells were previously exposed to LB95. In the cytotoxicity assays, the cell-free supernatant of the co-culture (CFSC) of VTEC with LB95 accounted for the lowest value obtained amongst the co-cultures of VTEC with LABs, and was significantly lower than the value obtained with the co-culture of VTEC with the two probiotic reference strains. The cytotoxicity of CFSC of VTEC with both LB95 and LB13 exhibited values not significantly different from the cell-free supernatant of the nonpathogenic E. coli B strain. Our results suggested that LB95 may be able to attenuate the virulence of Gram-positive and Gram-negative food-borne pathogens; together with other reported features of these strains, our data reveal their possible use in probiotic foods due to their interesting potential in preventing enteric infections in humans.

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Inhibition of Streptococcus pneumoniae adherence to human epithelial cells in vitro by the probiotic Lactobacillus rhamnosus GG

Cited by 37 publications

References 46 publications

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Labneh with probiotic properties produced from kefir: development and sensory evaluation

Lactobacillus plantarum LB95 impairs the virulence potential of Gram-positive and Gram-negative food-borne pathogens in HT-29 and Vero cell cultures

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