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LPS-induced chemokine production of human DC is suppressed by VSL#3. A set of genes representing cluster II was confirmed at the protein level. Supernatants were harvested from replicate DC cultures after 48 hours after stimulation and performed in parallel with the cultures used for mRNA profiling. Levels of various cytokines and chemokines were determined by multiplex bead array technology. Results are expressed as pg/ml ± SD. Rank-Wilcoxon paired test: * p<0.05, ** p<0.01, *** p<0.0001.
Source publication
VSL#3, a mixture of 8 different probiotic bacteria, has successfully been used in the clinic to treat Ulcerative Colitis. We previously identified the modulation of chemokines as a major mechanism in the protective effect of the VSL#3 in a mouse model of colitis. This was supported by in
vitro studies that implicated a role for VSL#3 in the suppres...
Citations
... Probiotics particularly Gram-positive members also conserve molecular patterns of microorganisms including lipoteichoic acids, exopolysaccharides, lipopolysaccharides, peptidoglycan, flagellin, the proteins of S and A layers, and the nucleic acids, which are recognized by recognition pattern of receptors. This can persuade a signalling of theimmunological cascades followed by a secretion of cytokines/interleukins, and other molecular effectors which are activators of animal immune functions resulting in induction of immunostimulatory or immunomodulatory effects (Remus et al., 2012;Soltani et al., 2019a;Zhou et al., 2019;Mariman et al., 2014). In addition, there are relations between tool-like cell receptor signalling-mediated recognition of probiotics and stimulation of the animal mucosal immunity (Vinderola et al., 2019;Galdeano et al., 2019). ...
... The pathogenesis of OLP is believed to involve a dysregulation of the immune system, (Jontell and Holmstrup, 2015;Zucoloto et al., 2019) including presentation of an unknown antigen, T lymphocyte activation and migration, production of proinflammatory cytokines and chemokines resulting in sub-epithelial inflammatory infiltrate, keratinocytes damage and disruption of epithelial homeostasis. (Ke et al., 2017;Lu et al., 2015;Marshall et al., 2017) Available evidence suggests that CXCL10 and IFNγ are key players in OLP-associated inflammation, (Tao et al., 2008) and previous in vitro studies showed that multi-strain probiotics are capable of suppressing LPS-induced chemokines, including CXCL10, through the blockade in the phosphorylation of the transcription factor STAT1. (Mariman et al., 2014) Clinically the disease presents with reticular hyperkeratotic changes of the oral mucosa, with more than 50% of the affected individuals also developing long-standing erosion and ulceration leading to reduced quality of life due to pain and dysfunction. (Osipoff et al., 2020) There remains no curative treatment and therefore management of OLP is aimed at reducing mucosal erosions/ulceration and the associated painful symptoms. ...
Objective
To preliminary evaluate the clinical effects of probiotics in individuals with symptomatic oral lichen planus, and the possible mechanisms of action.
Subjects and Methods
30 individuals with symptomatic oral lichen planus were recruited in a double-blind parallel group randomised controlled (1:1) proof-of-concept pilot trial of probiotic VSL#3 vs placebo. Efficacy outcomes included changes in pain numeric rating scale, oral disease severity score and the chronic oral mucosal disease questionnaire. Adverse effects, home diary, and withdrawals were assessed as feasibility outcomes. Mechanistic outcomes included changes in salivary and serum levels of CXCL10 and IFN-γ and in oral microbial composition.
Results
The probiotic VSL#3 was safe and well tolerated. We observed no statistically significant change in pain, disease activity, quality of life, serum/salivary CXCL10 or oral microbial composition with respect to placebo. Salivary IFN-γ levels demonstrate a trend for a reduced level in the active group (p=0.082) after 30 days of probiotic consumption.
Conclusions
The present proof-of-concept study provides some weak not convincing indication of biological and clinical effects of probiotic VSL#3 in individuals with painful oral lichen planus. Further research in this field is needed, with the current study providing useful information to the design of future clinical trials.
... VSL#3 also up-regulates the peroxisome proliferator-activated receptor α (PPARα) signaling pathway to antagonize the NF-κB pathway [32] . An appropriate dose of VSL#3 can induce the maturation of dendrite cells (DC) [27,28] , and VSL#3 can inhibit interferon-inducible protein-10 (IP-10) in intestinal epithelial cells (IEC) [22][23][24] and the lipopolysaccharide (LPS)-induced expression of chemokines (CXCL9, CXCL10, CCL2, CCL7, and CCL8) by inhibiting STAT-1 phosphorylation [27] . VSL#3 is also able to decrease interleukin (IL)-12 (p40) production induced by LPS [30] . ...
... VSL#3 also up-regulates the peroxisome proliferator-activated receptor α (PPARα) signaling pathway to antagonize the NF-κB pathway [32] . An appropriate dose of VSL#3 can induce the maturation of dendrite cells (DC) [27,28] , and VSL#3 can inhibit interferon-inducible protein-10 (IP-10) in intestinal epithelial cells (IEC) [22][23][24] and the lipopolysaccharide (LPS)-induced expression of chemokines (CXCL9, CXCL10, CCL2, CCL7, and CCL8) by inhibiting STAT-1 phosphorylation [27] . VSL#3 is also able to decrease interleukin (IL)-12 (p40) production induced by LPS [30] . ...
... Dendrite cells (DC) are the key cells in the production of immune responses and the modulation of inflammatory signaling pathways. The stimulation of human DC with an appropriate dose of VSL#3 will induce the maturation of immature DC [27,28] . However, VSL#3 (10 5 organisms/mL) did not change the immature phenotype and costimulatory molecule expression of DC, and VSL#3 (10 7 organisms/mL) was shown to increase the expression of CD40, CD80, CD86, and major histocompatibility complex (MHC) class II I-Ad during the last 3 d of culture in bone marrow-derived dendritic cells [29] . ...
Probiotics are known as "live microorganisms" and have been proven to have a health effect on hosts at the proper dose. Recently, a kind of probiotic mixture including eight live bacterial strains, VSL#3, has attracted considerable attention for its combined effect. VSL#3 is the only probiotic considered as a kind of medical food; it mainly participates in the regulation of the intestinal barrier function, including improving tight junction protein function, balancing intestinal microbial composition, regulating immune-related cytokine expression and so on. The objective of this review is to discuss the treatment action and mechanism for the administration of VSL#3 in chronic diseases of animals and humans (including children). We found that VSL#3 has a therapeutic or preventive effect in various systemic diseases per a large number of studies, including digestive systemic diseases (gastrointestinal diseases and hepatic diseases), obesity and diabetes, allergic diseases, nervous systemic diseases, atherosclerosis, bone diseases, and female reproductive systemic diseases.
... In these two-dimensional (2D) models, the interaction with the immune system or other tissues cannot be studied. Although the direct effect of VSL#3 was tested on spleen and dendritic cells (145,146), tissue-tissue interactions were lacking in these models. This problem can be (partially) solved in transwell coculture models, in which bacteria, mucosal immune cells, and intestinal epithelial cells can be studied together (147). ...
The human intestinal ecosystem is characterized by a complex interplay between different microorganisms and the host. The high variation within the human population further complicates the quest toward an adequate understanding of this complex system that is so relevant to human health and well-being. To study host-microbe interactions, defined synthetic bacterial communities have been introduced in gnotobiotic animals or in sophisticated in vitro cell models. This review reinforces that our limited understanding has often hampered the appropriate design of defined communities that represent the human gut microbiota. On top of this, some communities have been applied to in vivo models that differ appreciably from the human host. In this review, the advantages and disadvantages of using defined microbial communities are outlined, and suggestions for future improvement of host-microbe interaction models are provided. With respect to the host, technological advances, such as the development of a gut-on-a-chip system and intestinal organoids, may contribute to more-accurate in vitro models of the human host. With respect to the microbiota, due to the increasing availability of representative cultured isolates and their genomic sequences, our understanding and controllability of the human gut “core microbiota” are likely to increase. Taken together, these advancements could further unravel the molecular mechanisms underlying the human gut microbiota superorganism. Such a gain of insight would provide a solid basis for the improvement of pre-, pro-, and synbiotics as well as the development of new therapeutic microbes.
... Recent studies revealed that probiotic strains differentially modulate the DC expression of cytokines [20][21][22]. Lactobacillus can down-regulate IL-12 expression in lipopolysaccharide (LPS)-induced intestinal DC [23], and Enterococcus faecalis inhibits the expression of IL-4 and costimulatory molecular CD83, CD86 to reduce the number of mature DCs [24]. In our experiment, we isolated and identified the DCs from white blood cells (WBCs) of grass carp spleen and head kidney, and collected the transcriptome data of dendritic cells following Bacillus subtilis stimulation. ...
Bacillus subtilis is a common group of probiotics that have been widely used in the feed industry as they can increase host resistance to pathogens and balance the immune response. However, the regulatory mechanism of Bacillus subtilis on the host immune system remains unclear in teleosts. In this study, we isolated and enriched dendritic cells from white blood cells (WBCs), and then stimulated them with Bacillus subtilis. Morphological features, specific biological functions, and authorized functional molecular markers were used in the identification of dendritic cells. Subsequently, we collected stimulated cells at 0, 4, and 18 h, and then constructed and sequenced the transcriptomic libraries. A transcriptome analysis showed that 2557 genes were up-regulated and 1708 were down-regulated at 4 h compared with the control group (|Fold Change| ≥ 4), and 1131 genes were up-regulated and 1769 were down-regulated between the cells collected at 18 h and 4 h (|Fold Change| ≥ 4). Gene Ontology (GO) annotations suggested many differentially expressed genes (DEGs) (p < 0.05 and |Fold Change| ≥ 4) were involved in immune-related biological functions including immune system progress, cytokine receptor binding, and cytokine binding. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the cytokine–cytokine receptor interaction pathways were significantly enriched at both time points (p < 0.05), which may play a key role in the response to stimulation. Furthermore, mRNA expression level examination of several pro-inflammatory cytokines and anti-inflammatory cytokines genes by quantitative real-time polymerase chain reaction (qRT-PCR) indicated that their expressions can be significantly increased in Bacillus subtili, which suggest that Bacillus subtilis can balance immune response and tolerance. This study provides dendritic cell (DC)-specific transcriptome data in grass carp by Bacillus subtilis stimulation, allowing us to illustrate the molecular mechanism of the DC-mediated immune response triggered by probiotics in grass carp.
... Our findings here further reinforce former data providing evidence for anti-inflammatory properties of VSL#3. For instance, VSL#3 has been shown to suppress MCP-1 production from human dendritic cells in vitro [46] and to down-regulate colonic MCP-1 mRNA expression also in vivo [47]. Moreover VSL#3 application could ameliorate recurrent Th1-mediated TNBS colitis in mice by inducing IL-10 and IL-10-dependent regulatory T cells expressing TGF-β [48]. ...
Background
The incidence of human Campylobacter jejuni infections is progressively increasing worldwide. Probiotic compounds might open up valuable tools to decrease pathogen burden and subsequent pro-inflammatory immune responses, but in vivo data are scarce.
Methods and results
Secondary abiotic mice generated by broad-spectrum antibiotic treatment were perorally challenged with the commercial probiotic compound VSL#3 consisting of Streptococcus thermophilus, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium infantis, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus delbrueckii ssp. bulgaricus) either 5 days before (i.e. prophylactic regimen) or after (i.e. therapeutic regimen) peroral C. jejuni strain 81–176 infection, and analyzed 3 weeks following the initial bacterial re-association. Upon challenge, mice were colonized with the probiotic bacteria and/or C. jejuni at comparable intestinal loads, but co-colonization did not result in reduction of the pathogen burden. Remarkably, prophylactic as well as therapeutic VSL#3 treatment of C. jejuni infected mice ameliorated intestinal apoptosis and pro-inflammatory immune responses as indicated by lower numbers of innate and adaptive immune cell populations in the murine colon upon probiotic prophylaxis or treatment and reduced colonic concentrations of pro-inflammatory mediators including IL-6 and MCP-1. Importantly, concentrations of anti-inflammatory mediators such as IL-10 were significantly elevated in the colon of probiotics treated mice as compared to untreated controls. Strikingly, prophylactic VSL#3 treatment attenuated C. jejuni induced systemic pro-inflammatory responses as indicated by less TNF and IL-12p70 secretion in the spleen of VSL#3 pre-treated as compared to non-treated mice.
Conclusion
Administration of probiotic formulations such as VSL#3 might open up valuable strategies for prophylaxis and/or treatment of C. jejuni induced intestinal and systemic sequelae in vivo by the suppression of pro-inflammatory and induction of anti-inflammatory responses.
... 47 Furthermore, a recent study identified that VSL#3 treatment of dendritic cells reduced lipopolysaccharide-induced STAT-1 phosphorylation and transcription of STAT-1 gene targets including various inflammatory chemokines. 48 This is in broad agreement with our observations that VSL#3 reduces STAT-1 phosphorylation by IFN-g in IEC, and our data are consistent with a role for TCPTP in mediating VSL#3 inhibition of STAT-1 activation in multiple cell types. ...
Background:
VSL#3 is a probiotic compound that has been used in the treatment of inflammatory bowel disease. T-cell protein tyrosine phosphatase (TCPTP) is the protein product of the inflammatory bowel disease candidate gene, PTPN2, and we have previously shown that it protects epithelial barrier function. The aim of this study was to investigate whether VSL#3 improves intestinal epithelial barrier function against the effects of the inflammatory bowel disease-associated proinflammatory cytokine, interferon-gamma (IFN-γ) through activation of TCPTP.
Methods:
Polarized monolayers of T84 intestinal epithelial cells were treated with increasing concentrations of VSL#3 to determine effects on TCPTP expression and enzymatic activity. Therapeutic effects of VSL#3 against barrier disruption by IFN-γ were measured by transepithelial electrical resistance and fluorescein isothiocyanate-dextran permeability. A novel TCPTP-deficient HT-29 intestinal epithelial cell line was generated to study the role of TCPTP in mediating the effects of VSL#3. Tight junction protein distribution was assessed with confocal microscopy.
Results:
VSL#3 increased TCPTP protein levels and enzymatic activity, correlating with a VSL#3-induced decrease in IFN-γ signaling. VSL#3 corrected the decrease in transepithelial electrical resistance and the increase in epithelial permeability induced by IFN-γ. Moreover, the restorative effect of VSL#3 against IFN-γ signaling, epithelial permeability defects, altered expression and localization of the tight junction proteins claudin-2, occludin, and zonula occludens-1, were not realized in stable TCPTP/(PTPN2)-deficient HT-29 intestinal epithelial cells.
Conclusions:
VSL#3 reduces IFN-γ signaling and IFN-γ-induced epithelial barrier defects in a TCPTP-dependent manner. These data point to a key role for TCPTP as a therapeutic target for restoration of barrier function using probiotics.
... Lipopolysaccharide (LPS) and D-galactosamine could induce the phosphorylation of STAT1 and enhance the transcription of CXCL9 leading to the enhancement of liver inflammation, and even liver apoptosis and injury [15]. In this process, VSL#3 (a mixture of eight different probiotic bacteria) can specifically reduce the phosphorylation of STAT-1 [15,16]. STAT-1 can also be activated by IL-27 accompanied with IFN-γ. ...
... In addition, our previous work by Xia et al. identified that HBx protein can induce the CXCL9 transcription by activating NF-κB that binds to its promoter, and CXCL9 promotes the migration of leukocytes in liver with HBV infection [18]. Unlike STAT1, phosphorylation of NF-κB could not be suppressed by VSL#3 [16]. STAT1 and NF-κB can cooperatively regulate the expression of CXCL9, and this transcriptional synergy could cause the enhanced recruitment of RNA polymerase II complex to the promoter via simultaneous interaction of CBP with STAT1 and NF-κB [10]. ...
Chemokines are a group of low molecular weight peptides. Their major function is the recruitment of leukocytes to inflammation sites, but they also play a key role in tumor growth, angiogenesis, and metastasis. In the last few years, accumulated experimental evidence supports that monokine induced by interferon (IFN)-gamma (CXCL9), a member of CXC chemokine family and known to attract CXCR3- (CXCR3-A and CXCR3-B) T lymphocytes, is involved in the pathogenesis of a variety of physiologic diseases during their initiation and their maintenance. This review for the first time presents the most comprehensive summary for the role of CXCL9 in different types of tumors, and demonstrates its contradictory role of CXCL9 in tumor progression. Altogether, this is a useful resource for researchers investigating therapeutic opportunities for cancer.
... Some in vitro studies have implicated that VSL#3 can suppress lipopolysaccharide-induced chemokine production by inhibiting signal transducer and activator of transcription 1 phosphorylation. 34 Mariman et al 35 studied C57BL/6 and BALB/c mice, and found that VSL#3 mediated both pro-and anti-inflammatory responses in bone marrow-derived dendritic cells. Dai et al 36 demonstrated that VSL#3 exerted antiinflammatory activity via the phosphoinositide 3kinase/protein kinase B and nuclear factor-kB pathway in a rat model of dextran sulfate sodiuminduced colitis. ...
Objective
To evaluate the maintenance effect of probiotics versus that of aminosalicylates on ulcerative colitis.
Methods
MEDLINE, EMBASE, the Cochrane Controlled Trials Register, and the Chinese Biomedical Database were searched in English or Chinese. Data extracted were selected with strict criteria.
Results
In six randomized controlled trials (RCTs), a total of 721 participants were enrolled and the maintenance effect of probiotics (n = 364) versus that of aminosalicylates (n = 357) on ulcerative colitis was investigated. No significant difference was observed between probiotics and aminosalicylate groups (relative risk (RR) = 1.08; 95% confidence interval (CI): 0.91–1.28; P = 0.40). Three RCTs compared the incidence of adverse events with probiotics versus those with aminosalicylates. No significant difference was observed in the incidence of adverse events between the two groups (RR = 1.20; 95% CI: 0.92–1.56; P = 0.17).
Conclusions
Probiotics and aminosalicylates both showed a maintenance effect on ulcerative colitis. However, more well-designed RCTs are required.
... The involvement of DCs and Treg cells in probioticmediated immune responses has been previously demonstrated (Drakes et al., 2004;Foligne et al., 2007;Mariman et al., 2014;Strisciuglio et al., 2015). For example, incubation with selected LAB strains induced partial maturation in DCs in vitro, and the transfer of these DCs conferred protection against TNBS-induced colitis and induced the production of CD4 + CD25 + Treg cells (Foligne et al., 2007). ...
Inflammatory bowel disease is a group of chronic, incurable inflammatory disorders of the gastrointestinal tract that cause severe diarrhoea, intestinal inflammation, pain, fatigue and weight loss. In this study, we first developed a model of Citrobacter rodentium-induced colitis and then evaluated the protective effects of selected probiotics on inflammation. The results showed that administration of a combination of probiotics including Lactobacillus rhamnosus ATCC 53103, Lactobacillus acidophilus ATCC 4356 and Lactobacillus plantarum A significantly increased the production of CD11c(+) dendritic cells in the spleen (3.62% vs phosphate buffered saline (PBS)-treated control, P<0.01) and mesenteric lymph nodes (MLNs). In addition, the presence of probiotics significantly up-regulated the development of CD4(+)/CD25(+)/Foxp3(+) regulatory T cells in MLNs by approximately 2.07% compared to the effect observed in the PBS-treated control (P<0.01) and down-regulated the expression of inflammatory cytokines, including interleukin-17, tumour necrosis factor-α and interferon-γ, by 0.11, 0.11 and 0.15%, respectively, compared to the effect observed in the PBS-treated control (P<0.01).These effects conferred protection against colitis, as shown by histopathological analyses.
