Unraveling Mechanisms of Action of Probiotics

Sherman, Philip M.; Ossa, Juan C.; Johnson‐Henry, Kathene C.

doi:10.1177/0884533608329231

Cited by 188 publications

(133 citation statements)

References 42 publications

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“…HIEC play a central role in absorptive and secretory functions, protecting the organisms from bacteria and toxins, providing an effective barrier against the complex antigenic load of intestinal contents (3,25). However, difficulties still exist in establishing an HIEC cell line in China.…”

Section: Discussionmentioning

confidence: 99%

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Liu

Zhang

Zhou

et al. 2010

Braz J Med Biol Res

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Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures) was enhanced to 60% of a total of 10 cultures (initiated from 8 distinct fetal small intestines), allowing the generation of viable epithelial cell cultures. Western blot, real-time PCR and immunofluorescent staining showed that cytokeratins 8, 18 and mouse intestinal mucosa-1/39 had high expression levels in human intestinal epithelial cells. Differentiated markers such as sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV also showed high expression levels in human intestinal epithelial cells. Differentiated human intestinal epithelial cells, with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa-1/39) and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV), may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. This is relatively simple, requiring no sophisticated techniques or instruments, and may have a number of varied applications.

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Section: Discussionmentioning

confidence: 99%

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Liu

Zhang

Zhou

et al. 2010

Braz J Med Biol Res

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“…Multiple mechanisms of action for the beneficial effect of probiotics have been proposed (Ng et al, 2009;Sherman et al, 2009;Lebeer et al, 2008). The ability to adhere to epithelial cells and mucosal surfaces has been suggested as being an important property of many bacterial strains used as probiotics.…”

Section: Introductionmentioning

confidence: 99%

“…Several workers have suggested that the ability of beneficial micro-organisms to aggregate and adhere aids in colonization of the gut and in the establishment of a barrier which prevents enteropathogens from establishing an infection. Factors which prevent infection include the physical presence of beneficial micro-organisms and modulation of the gut immune system by these organisms (Cesena et al, 2001;Voltan et al, 2007;Collado et al, 2008;Sherman et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

In vitro characterization of aggregation and adhesion properties of viable and heat-killed forms of two probiotic Lactobacillus strains and interaction with foodborne zoonotic bacteria, especially Campylobacter jejuni

Tareb

Bernardeau²,

Guéguen

et al. 2013

Journal of Medical Microbiology

113

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Bacterial aggregation and/or adhesion are key factors for colonization of the digestive ecosystem and the ability of probiotic strains to exclude pathogens. In the present study, two probiotic strains, Lactobacillus rhamnosus CNCM-I-3698 and Lactobacillus farciminis CNCM-I-3699, were evaluated as viable or heat-killed forms and compared with probiotic reference Lactobacillus strains (Lb. rhamnosus GG and Lb. farciminis CIP 103136). The autoaggregation potential of both forms was higher than that of reference strains and twice that of pathogenic strains. The coaggregation potential of these two beneficial micro-organisms was evaluated against several pathogenic agents that threaten the global safety of the feed/food chain: Escherichia coli, Salmonella spp., Campylobacter spp. and Listeria monocytogenes. The strongest coaggregative interactions were demonstrated with Campylobacter spp. by a coaggregation test, confirmed by electron microscopic examination for the two forms. Viable forms were investigated for the nature of the bacterial cell-surface molecules involved, by sugar reversal tests and chemical and enzymic pretreatments. The results suggest that the coaggregation between both probiotic strains and C. jejuni CIP 70.2 T is mediated by a carbohydrate-lectin interaction. The autoaggregation potential of the two probiotics decreased upon exposure to proteinase, SDS or LiCl, showing that proteinaceous components on the surface of the two lactobacilli play an important role in this interaction. Adhesion abilities of both Lactobacillus strains were also demonstrated at significant levels on Caco-2 cells, mucin and extracellular matrix material. Both viable and heat-killed forms of the two probiotic lactobacilli inhibited the attachment of C. jejuni CIP 70.2 T to mucin. In conclusion, in vitro assays showed that Lb. rhamnosus CNCM-I-3698 and Lb. farciminis CNCM-I-3699, as viable or heat-killed forms, are adherent to different intestinal matrix models and are highly aggregative in vitro with pathogens, especially Campylobacter spp., the most commonly reported zoonotic agent in the European Union. This study supports the need for further in vivo investigations to demonstrate the potential food safety benefits of Lb. rhamnosus CNCM-I-3698 and Lb. farciminis CNCM-I-3699, live or heat-killed, in the global feed/food chain.

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“…The interaction between intestinal epithelial cells and microorganisms is a complex process (Sherman et al 2009;Di Mauro et al 2013). A toll-like receptor (TLR) family on intestinal epithelial cells plays an important role in recognizing the intestine foreigners, which subsequently initiates the inflammation response Kawai & Akira 2009;O'Neill et al 2009;Lavelle et al 2010).…”

Section: Introductionmentioning

confidence: 99%

The inflammation regulation effects ofEnterococcus faeciumHDRsEf1 on human enterocyte-like HT-29 cells

Tian

Yang

et al. 2016

Animal Cells and Systems

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Enterococcus faecium HDRsEf1 strain used as a probiotic to inhibit intestine inflammation and improve animal growth performance has been proved by our research team; however, it remains unclear how HDRsEf1 was recognized by intestine cells and how it activates the downstream pathway which benefit intestine health. In this study, HDRsEf1 was used to stimulate HT-29 cell line to partially uncover the intestine benefit mechanism of HDRsEf1. The results of cell viability assays showed that HDRsEf1 had no toxicity on HT-29 at concentrations up to 1 × 10 8 CFU/mL, HDRsEf1 could upregulate the TLR1, TLR2, and TLR6 mRNA level, especially TLR2, and significantly downregulate the mRNA level of TLR4, TLR5, TLR7, TLR8, but did not significantly affect the mRNA or protein level of MyD88, which suggests that HDRsEf1 activates the TLR2 pathway in an MyD88-independent pattern. HDRsEf1 could significantly downregulate the mRNA level of pro-inflammatory factors IL-1β, IL-6, IL-8, IL-12p35, IL-17, and TNF-α and did not affect the anti-inflammatory factors IL-10, PPAR-γ, and TSLP; besides HDRsEf1 did not upregulate the degradation of IκB in HT-29 cells. By contrast, enterohemorrhagic E. coli (EHEC) O157:H7 strongly up-regulated the mRNA level of pro-inflammatory factors IL-1β, IL-6, IL-8, IL-23, and TNF-α, downregulated obviously anti-inflammatory factor PPAR-ɣ, and obviously upregulated the degradation of IκB, which suggested that HDRsEf1 may act as an antagonist to regulate intestine inflammation response to intestine pathogen. These findings shed a light on the intestine benefit mechanism of HDRsEf1. ARTICLE HISTORY

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Unraveling Mechanisms of Action of Probiotics

Cited by 188 publications

References 42 publications

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

In vitro characterization of aggregation and adhesion properties of viable and heat-killed forms of two probiotic Lactobacillus strains and interaction with foodborne zoonotic bacteria, especially Campylobacter jejuni

The inflammation regulation effects ofEnterococcus faeciumHDRsEf1 on human enterocyte-like HT-29 cells

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