Critical Adverse Impact of IL-6 in Acute Pneumovirus Infection. Caroline M Percopo;Michelle Ma;Todd A Brenner;Julia O Krumholz;Timothy J Break;Karen Laky;Helene F Rosenberg. 2018. J Immunol. 202. PMID: 30578308

Severe respiratory virus infections feature robust local host responses that contribute to disease severity. Immunomodulatory strategies that limit virus-induced inflammation may be of critical importance, notably in the absence of antiviral vaccines. In this study, we examined the role of the pleiotropic cytokine IL-6 in acute infection with pneumonia virus of mice (PVM), a natural rodent pathogen that is related to respiratory syncytial virus and that generates local inflammation as a feature of severe infection. In contrast to Influenza A, PVM is substantially less lethal in IL-6 -/- mice than it is in wild-type, a finding associated with diminished neutrophil recruitment and reduced fluid accumulation in lung tissue. Ly6Chi proinflammatory monocytes are recruited in response to PVM via a CCR2-dependent mechanism, but they are not a major source of IL-6 nor do they contribute to lethal sequelae of infection. By contrast, alveolar macrophages are readily infected with PVM in vivo; ablation of alveolar macrophages results in prolonged survival in association with a reduction in virus-induced IL-6. Finally, as shown previously, administration of immunobiotic Lactobacillus plantarum to the respiratory tracts of PVM-infected mice promoted survival in association with diminished levels of IL-6. We demonstrated in this study that IL-6 suppression is a critical feature of the protective mechanism; PVM-infected IL-6 -/- mice responded to low doses of L. plantarum, and administration of IL-6 overcame L. plantarum-mediated protection in PVM-infected wild-type mice. Taken together, these results connect the actions of IL-6 to PVM pathogenesis and suggest cytokine blockade as a potential therapeutic modality in severe infection.
Lipoteichoic Acid Is Involved in the Ability of the Immunobiotic Strain Lactobacillus plantarum CRL1506 to Modulate the Intestinal Antiviral Innate Immunity Triggered by TLR3 Activation. Hiroya Mizuno;Lorena Arce;Kae Tomotsune;Leonardo Albarracin;Ryutaro Funabashi;Daniela Vera;Md Aminul Islam;Maria Guadalupe Vizoso-Pinto;Hideki Takahashi;Yasuko Sasaki;Haruki Kitazawa;Julio Villena. 2020. Front Immunol. 11. PMID: 32328062

Studies have demonstrated that lipoteichoic acid (LTA) is involved in the immunomodulatory properties of some immunobiotic lactobacilli. The aim of this work was to evaluate whether LTA contributes to the capacity of Lactobacillus plantarum CRL1506 in modulating the intestinal innate antiviral immune response. A D-alanyl-lipoteichoic acid biosynthesis protein (dltD) knockout CRL1506 strain (L. plantarumΔdltD) was obtained, and its ability to modulate Toll-like receptor (TLR)-3-mediated immune response was evaluated in vitro in porcine intestinal epithelial (PIE) cells and in vivo in Balb/c mice. Wild-type (WT) CRL1506 (L. plantarum WT) was used as positive control. The challenge of PIE cells with the TLR3 agonist poly(I:C) significantly increased interferon (IFN)-β, interleukin (IL)-6, and monocyte chemoattractant protein (MCP)-1 expressions. PIE cells pretreated with L. plantarumΔdltD or L. plantarum WT showed higher levels of IFN-β while only L. plantarum WT significantly reduced the expression of IL-6 and MCP-1 when compared with poly(I:C)-treated control cells. The oral administration of L. plantarum WT to mice prior the intraperitoneal injection of poly(I:C) significantly increased IFN-β and IL-10 and reduced intraepithelial lymphocytes (CD3+NK1.1+CD8αα+) and pro-inflammatory mediators (TNF-α, IL-6, and IL-15) in the intestinal mucosa. Similar to the WT strain, L. plantarumΔdltD-treated mice showed enhanced levels of IFN-β after poly(I:C) challenge. However, treatment of mice with L. plantarumΔdltD was not able to increase IL-10 or reduce CD3+NK1.1+CD8αα+ cells, TNF-α, IL-6, or IL-15 in the intestine. These results indicate that LTA would be a key molecule in the anti-inflammatory effect induced by the CRL1506 strain in the context of TLR3-mediated inflammation.
Probiotics potentiate IL-6 production in IL-1beta-treated Caco-2 cells through a heat shock-dependent mechanism. Natasha Reilly;Vitaliy Poylin;Michael Menconi;Andrew Onderdonk;Stig Bengmark;Per-Olof Hasselgren. 2007. Am J Physiol Regul Integr Comp Physiol. 293. PMID: 17634198

IL-6 may exert anti-inflammatory and protective effects in intestinal mucosa and enterocytes. The influence of probiotics on mucosal and enterocyte IL-6 production is not known. We tested the hypothesis that the probiotic bacteria Lactobacillus paracasei and Lactobacillus plantarum regulate IL-6 production in intestinal epithelial cells. Cultured Caco-2 cells were treated with 1 ng/ml of IL-1beta in the absence or presence of different concentrations of L. paracasei or L. plantarum followed by measurement of IL-6 production. The role of heat shock response was examined by determining the expression of heat shock protein 70 (hsp70) and hsp27, by downregulating their expression with small interfering RNA (siRNA), or by treating cells with quercetin. Treatment of the Caco-2 cells with IL-1beta resulted in increased IL-6 production, confirming previous reports from this laboratory. Probiotics alone did not influence IL-6 production, but the addition of probitoics to IL-1beta-treated cells resulted in a substantial augmentation of IL-6 production. Treatment of the Caco-2 cells with live L. paracasei increased cellular levels of hsp70 and hsp27 and the potentiating effect on IL-6 production was inhibited by quercetin and by hsp70 or hsp27 siRNA. Results suggest that probiotics may enhance IL-6 production in enterocytes subjected to an inflammatory stimulus and that this effect may, at least in part, be heat shock dependent.
Effects of Lactobacillus plantarum 2142 and sodium n-butyrate in lipopolysaccharide-triggered inflammation: comparison of a porcine intestinal epithelial cell line and primary hepatocyte monocultures with a porcine enterohepatic co-culture system. O Farkas;G Mátis;E Pászti-Gere;O Palócz;A Kulcsár;J Petrilla;Gy Csikó;Zs Neogrády;P Gálfi. 2014. J Anim Sci. 92. PMID: 24987069

This study was based on our previously developed double-layered enterohepatic co-culture system, composed of nontumorigenic porcine intestinal epithelial cell line (IPEC-J2) and primary culture of porcine hepatocytes. The anti-inflammatory effect of spent culture supernatant of Lactobacillus plantarum 2142 (Lp2142; 13.3%) and sodium n-butyrate (2 mM) was tested on IPEC-J2 and hepatocyte monocultures as well as on the gut-liver co-culture. To mimic inflammation, lipopolysaccharide (LPS; 1 and 10 μg/mL) was applied. Production of IL-8 and IL-6 was measured as a marker of inflammatory responses. The paracellular permeability of the intestinal epithelium was also monitored by fluoresceinisothiocyanate-labeled dextran 4 assay. Significant increase of IL-8 concentration was observed in the IPEC-J2 monoculture (P < 0.01) while the level of IL-6 was not changed following LPS treatment. Concentration of IL-8 and IL-6 was grown significantly in hepatocyte monocultures (P < 0.05 and P < 0.001) as well as in the co-culture after 10 μg/mL LPS treatment (P < 0.001 and P < 0.001). One microgram per milliliter LPS caused elevated IL-8 level in the co-culture (P < 0.001) and in the hepatocyte monoculture (P < 0.01), while it caused increased IL-6 level only in the hepatocytes (P < 0.001). Production of IL-8 was significantly decreased by butyrate in case of 1 μg/mL as well as 10 μg/mL LPS exposure in the co-culture (P < 0.001). Application of butyrate also reduced IL-6 level in the co-culture after 10 μg/mL LPS treatment (P < 0.01). Lactobacillus plantarum 2142 decreased IL-8 level after incubation with 1 μg/mL LPS (P < 0.001), while in case of 10 μg/mL LPS treatment only a marginal lowering in IL-8 (P = 0.064) release was measured. The IL-6 concentration was significantly reduced (P < 0.01 in case of 1 μg/mL LPS treatment) by Lp2142 in the co-culture. Contrarily, the elevated IL-8 and IL-6 level of hepatocytes has not been reduced in case of either butyrate or Lp2142 addition. The enterohepatic co-culture model offers a possibility for fast and reliable screening of new candidates against enteric inflammation, which are of special interest in porcine medicine and health management. According to our results, Lp2142 and butyrate both seem to be effective as anti-inflammatory agents in LPS-triggered inflammatory response, tested in the gut-liver co-culture model.
Lactobacillus plantarum prevents and mitigates alcohol-induced disruption of colonic epithelial tight junctions, endotoxemia, and liver damage by an EGF receptor-dependent mechanism. Pradeep K Shukla;Avtar S Meena;Bhargavi Manda;Maria Gomes-Solecki;Paula Dietrich;Ioannis Dragatsis;RadhaKrishna Rao. 2018. FASEB J. . PMID: 29912589

Pathogenesis of alcohol-related diseases such as alcoholic hepatitis involves gut barrier dysfunction, endotoxemia, and toxin-mediated cellular injury. Here we show that Lactobacillus plantarum not only blocks but also mitigates ethanol (EtOH)-induced gut and liver damage in mice. L. plantarum blocks EtOH-induced protein thiol oxidation, and down-regulation of antioxidant gene expression in colon L. plantarum also blocks EtOH-induced expression of TNF-α, IL-1β, IL-6, monocyte chemotactic protein 1 ( MCP1), C-X-C motif chemokine ligand ( CXCL)1, and CXCL2 genes in colon. Epidermal growth factor receptor (EGFR) signaling mediates the L. plantarum-mediated protection of tight junctions (TJs) and barrier function from acetaldehyde, the EtOH metabolite, in Caco-2 cell monolayers. In mice, doxycycline-mediated expression of dominant negative EGFR blocks L. plantarum-mediated prevention of EtOH-induced TJ disruption, mucosal barrier dysfunction, oxidative stress, and inflammatory response in colon. L. plantarum blocks EtOH-induced endotoxemia as well as EtOH-induced pathologic lesions, triglyceride deposition, oxidative stress, and inflammatory responses in the liver by an EGFR-dependent mechanism. L. plantarum treatment after injury accelerated recovery from EtOH-induced TJ, barrier dysfunction, oxidative stress, and inflammatory response in colon, endotoxemia, and liver damage. Results demonstrate that L. plantarum has both preventive and therapeutic values in treatment of alcohol-induced tissue injury, particularly in alcoholic hepatitis.-Shukla, P. K., Meena, A. S., Manda, B., Gomes-Solecki, M., Dietrich, P., Dragatsis, I., Rao, R. Lactobacillus plantarum prevents and mitigates alcohol-induced disruption of colonic epithelial tight junctions, endotoxemia, and liver damage by an EGF receptor-dependent mechanism.
Lactobacillus plantarum C88 protects against aflatoxin B1-induced liver injury in mice via inhibition of NF-κB-mediated inflammatory responses and excessive apoptosis. Li Huang;Zijian Zhao;Cuicui Duan;Chao Wang;Yujuan Zhao;Ge Yang;Lei Gao;Chunhua Niu;Jingbo Xu;Shengyu Li. 2019. BMC Microbiol. 19. PMID: 31357935

BACKGROUND: Probiotics play an important role in the human and animal defense against liver damage. However, the protective mechanism of Lactobacillus plantarum C88 on chronic liver injury induced by mycotoxin remains unclear. RESULTS: In this study, the addition of L. plantarum C88 obviously ameliorated the increased contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total cholesterol and triglyceride, the diminish contents of total protein and albumin in serum of mice challenged with AFB1. Simultaneously, L. plantarum C88 attenuated the inflammatory response via significantly reducing the levels of pro-inflammatory factors, including interleukin-1β (IL-1β), IL-6, IL-8, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in serum. Furthermore, L. plantarum C88 remarkably down-regulated the nuclear factor kappa B (NF-κB) signaling pathways by weakening the expression of toll-like receptor 2 (TLR2) and TLR4, and inhibited NF-κB nuclear translocation through enhancing the expression of NF-κB inhibitor (IκB). Neutralization experiments confirmed that L. plantarum C88 decreased the levels of some pro-inflammatory factors due to the suppression of the NF-κB signaling pathways. Besides, L. plantarum C88 decreased the levels of Bax and Caspase-3, elevated the level of Bcl-2, and reduced mRNA expressions of Fatty acid synthetase receptor (Fas), FAS-associated death domain (FADD), TNF receptor associated death domain (TRADD) and Caspase-8 in the liver. CONCLUSIONS: Probiotic L. plantarum C88 prevented AFB1-induced secretion of pro-inflammatory cytokines by modulating TLR2/NF-κB and TLR4/NF-κB pathways. The molecular mechanisms of L. plantarum C88 in ameliorating AFB1-induced excessive apoptosis included regulating the mitochondrial pathway and cell death receptor pathways.
Identification of the key physiological characteristics of Lactobacillus plantarum strains for ulcerative colitis alleviation. Yang Liu;Yingyue Sheng;Qiqi Pan;Yuzheng Xue;Leilei Yu;Fengwei Tian;Jianxin Zhao;Hao Zhang;Qixiao Zhai;Wei Chen. 2020. Food Funct. 11. PMID: 31984399

Lactobacillus plantarum is a probiotic that is widely used to prevent ulcerative colitis (UC). However, the effects of this species are strain-specific. We believe that the physiological characteristics of L. plantarum strains may affect their UC-alleviating function. Therefore, this study investigated the relationship between the alleviating effect of L. plantarum strains on UC and their physiological characteristics in vitro. The physiological characteristics of 14 L. plantarum strains were assayed in vitro, including gastrointestinal transit tolerance, oligosaccharide fermentation, HT-29 cell adhesion, generation time, exopolysaccharide production, acetic acid production, and conjugated linoleic acid (CLA) synthesis. To create animal models, colitis was established in C57BL/6 mice by adding 3.5% dextran sulfate sodium to drinking water for 7 days. L. plantarum strains with significantly different physiological characteristics were orally administered to the mice at a dose of 3 × 109 CFU. The results indicated that among the tested L. plantarum strains, L. plantarum N13 and L. plantarum CCFM8610 significantly alleviated colitis in the mice, as observed from the restoration of the body weight and disease activity index (DAI) score, recovery of the gut microbiota composition, reduced expression of pro-inflammatory cytokines, and significantly inhibited expression of p65. Correlation analysis indicated that four of the measured physiological characteristics (gastrointestinal transit tolerance, HT-29 cell adhesion, generation time, and CLA synthesis) were related to the UC-alleviating effects to different degrees. The strongest correlation was observed between the CLA synthesis ability and UC-alleviating effects (with Pearson correlation coefficients for IL-1β, IL-6, IL-17F, TNF-α, myeloperoxidase, and the DAI all below -0.95). The ability to synthesize CLA may be the key physiological characteristic of L. plantarum in UC alleviation. Our findings may contribute to the rapid screening of lactic acid bacterial strains with UC-alleviating effects.
[Lactobacillus plantarum NDC 75017 affects il-6 gene expression in Caco-2 cells]. Ying Liu;Chaoxin Man;Xuena Lv;Linlin Han;Yuqing Xue;Xiangyi Yang;Yi Shan;Yinghua Zhang;Yujun Jiang. 2013. Wei Sheng Wu Xue Bao. 52. PMID: 23289322

OBJECTIVE: We tested the expression of immune-related gene interleukin 6 (il-6) in vitro to understand the influence from Lactobacillus plantarum NDC 75017 on host cells and further to reveal the regulatory mechanism. METHODS: Caco-2 cells were cocultured with L. plantarum NDC 75017 for 0, 2, 4, 6, 8, 10 and 12 h, the total RNA were extracted; then the expressions of il-6 and tlr2 genes were analyzed by Real Time RT-PCR. The phosphorylation level of NF-KB was analyzed by Western Blot after the Caco-2 cells stimulation with L. plantarum NDC 75017 at 0, 0.5, 1, 2 and 4 h. Caco-2 cells were pretreated with pyrrolidine dithiocarbamate for 30 min before being treated with L. plantarum NDC 75017 for 2 h, then the total RNA was extracted and the expressions of il-6 and tlr2 genes were analyzed by Real Time RT-PCR. RESULTS: Lactobacillus plantarum NDC 75017 could induce the expressions of il-6 and tlr2 in Caco-2 cells, the il-6 and tlr2 expressions peaked at 8 h and 6 h after cocultured with L. plantarum NDC 75017. L. plantarum NDC 75017 could rapidly activate the phosphorylation of NF-kappaB, and the expressions of il-6 and tlr2 were decreased notably after pretreated with pyrrolidine dithiocarbamate. CONCLUSION: L. plantarum NDC 75017 could up-regulate and then down-regulate the expression of il6 through rapidly activating tlr2-mediated NF-kappaB signaling pathway in Caco-2 cells.
Bacteriocin biosynthesis contributes to the anti-inflammatory capacities of probiotic Lactobacillus plantarum. X Yin;D Heeney;Y Srisengfa;B Golomb;S Griffey;M Marco. 2017. Benef Microbes. 9. PMID: 29065706

Plantaricin EF (PlnEF) is a class IIb bacteriocin produced by Lactobacillus plantarum. We compared L. plantarum NCIMB8826 and LM0419, a plnEFI deletion mutant of that strain lacking plnEF and the gene for the cognate immunity protein plnI, in a 2,4,6-trinitrobenzenesulfonic acid (TNBS) induced mouse model of acute inflammatory bowel disease. Mice fed either L. plantarum NCIMB8826 or LM0419 were not protected against TNBS according to either disease activity or histology (Ameho) scores. Mice consuming NCIMB8826 exhibited intermediate (non-significant) levels of colonic tumour necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) that ranged between the TNBS-treated animals and healthy controls. By comparison, TNF-α and IL-6 quantities were elevated in mice given L. plantarum LM0419 and equivalent to mice given TNBS alone. Both strains survived digestive tract transit in equal numbers and did not result in global changes to the bacterial composition in the intestine according to 16S rRNA gene sequencing either prior to or after TNBS administration. Examination of intestinal taxa showed that mice consuming wild-type L. plantarum, but not LM0419 contained lower proportions of Mucispirillum (Deferribacteres phylum) in the faeces prior to TNBS administration and Parabacteroides (Bacteroidetes phylum) in the caecum after disease induction. Parabacteroides also positively correlated with disease activity and histology scores. These findings suggest a role for PlnEFI production by L. plantarum in benefiting digestive tract health.
Effects of Lactobacillus Plantarum and Lactobacillus Helveticus on Renal Insulin Signaling, Inflammatory Markers, and Glucose Transporters in High-Fructose-Fed Rats. Omer A Korkmaz;Esra Sumlu;H Bugra Koca;M Bilgehan Pektas;Aytac Kocabas;Gokhan Sadi;Fatma Akar. 2019. Medicina (Kaunas). 55. PMID: 31137715

Background and Objectives: The excess consumption of fructose in the diet may cause metabolic syndrome, which is associated with an increased risk of kidney disease. There is limited data on probiotic treatment in high-fructose-induced metabolic syndrome. The present study aims to investigate whether the supplementation of Lactobacillus plantarum (L. plantarum) and Lactobacillus helveticus (L. helveticus) could provide an improving effect on the renal insulin signaling effectors, inflammatory parameters, and glucose transporters in fructose-fed rats. Materials and Methods: The model of metabolic syndrome in male Wistar rats was produced by fructose, which was given as 20% solution in drinking water for 15 weeks. L. plantarum and L. helveticus supplementations were given by gastric gavage from 10 to 15 weeks of age. Results: High-fructose consumption in rats reduced renal protein expressions of insulin receptor substrate (IRS)-1, protein kinase B (AKT), and endothelial nitric oxide synthase (eNOS), which were improved by L. plantarum and partially by L. helveticus supplementations. Dietary fructose-induced elevations in renal tissue levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6, and IL-10, as well as expression of IL-6 mRNA, were attenuated, especially in L. plantarum treated rats. The increased renal expression of sodium-glucose cotransporter-2 (SGLT2), but not that of glucose transporter type-5 (GLUT5), was suppressed by the treatment with L. plantarum. Conclusion: Suppression in insulin signaling pathway together with the induction of inflammatory markers and upregulation of SGLT2 in fructose-fed rats were improved by L. plantarum supplementation. These findings may offer a new approach to the management of renal dysregulation induced by dietary high-fructose.
Immunomodulatory Effects of Lactobacillus plantarum on Inflammatory Response Induced by Klebsiella pneumoniae. Marjolaine Vareille-Delarbre;Sylvie Miquel;Sophie Garcin;Thomas Bertran;Damien Balestrino;Bertrand Evrard;Christiane Forestier. 2019. Infect Immun. 87. PMID: 31481408

Some respiratory infections have been associated with dysbiosis of the intestinal microbiota. The underlying mechanism is incompletely understood, but cross talk between the intestinal microbiota and local immune cells could influence the immune response at distal mucosal sites. This has led to the concept of enhancing respiratory defenses by modulating the intestinal microbiota with exogenous supplementation of beneficial strains. In this study, we examined the effect of Lactobacillus plantarum CIRM653 on the inflammatory response induced by the pathogen Klebsiella pneumoniae Oral administration of L. plantarum CIRM653 to mice subsequently infected by K. pneumoniae via the nasal route (i) reduced the pulmonary inflammation response, with decreased numbers of lung innate immune cells (macrophages and neutrophils) and cytokines (mouse keratinocyte-derived chemokine [KC], interleukin-6 [IL-6], and tumor necrosis factor alpha [TNF-α]) in the bronchoalveolar fluid, and (ii) induced an immunosuppressive Treg response in lungs. In vitro coincubation of L. plantarum CIRM653 and K. pneumoniae with human dendritic cells and peripheral blood mononuclear cells resulted in decreased Th1 (IL-12p70 and interferon gamma [IFN-γ]) and Th17 (IL-23 and IL-17) and increased Treg (IL-10) cytokine levels compared to those observed for K. pneumoniae-infected cells. Neither K. pneumoniae nor L. plantarum CIRM653 had any effect on cytokine production by intestinal epithelial cells in vitro, but the induction of the NF-κB pathway and IL-8 and IL-6 production by K. pneumoniae in airway epithelial cells was significantly reduced when the pathogen was coincubated with L. plantarum CIRM653. The remote IL-10-mediated modulation of the K. pneumoniae inflammatory response by L. plantarum CIRM653 supports the concept of immunomodulation by beneficial bacteria through the gut-lung axis.
Preventive use of Lactobacillus plantarum LS/07 and inulin to relieve symptoms of acute colitis. Emília Hijová;Alena Šoltésová;Rastislav Salaj;Jozef Kuzma;Ladislav Strojný;Alojz Bomba;Kristína Gregová. 2015. Acta Biochim Pol. 62. PMID: 26345092

The aim of presented study was to investigate the influence of Lactobacillus plantarum LS/07 and inulin on the activity of β-glucuronidase enzyme, and counts of coliform and lactobacilli in fresh caecal digesta, cytokine levels (IL-6, IL-8), and trancription nuclear factor kappa beta (NFκB) activities in colon tissue and blood samples of rats with dextran sulphate sodium (DSS) induced acute colitis. The rats were randomly divided into four groups - CG, AC, AC+PRE and AC+PRO. Colitis was induced using of 5% DSS in drinking water for 7d. DSS application increased activity of β-glucuronidase (P < 0.001), increased counts of coliforms, and decreased lactobacilli counts (P < 0.05) in comparison to control group. Serum and tissue levels of IL-6 and IL-8 as well as tissue NFκB activities showed increased expression in acute colitis group. Inulin diet modified counts of microorganims and decreased β-glucuronidase activity, suppressed NFκB activities (P < 0.001) and down regulate synthesis of IL-6 (P < 0.01) in serum and colon tissue and tissue IL-8 (P < 0.05). Lactobacillus plantarum LS/07 decreased β-glucuronidase activity (P < 0.05), levels of IL-6 and IL-8 (P < 0.001). These results were consistent with the addition of histological findings. Our results indicate that dietary intake of Lactobacillus plantarum LS/07 and inulin suppressed expression observed markers, which play an important role in the inflammatory process, which predisposes their use in prevention or treatment of acute colitis.
The anti-allergic activity of Lactobacillus plantarum L67 and its application to yogurt. Sooyeon Song;Sei-Jung Lee;Dong-June Park;Sejong Oh;Kye-Taek Lim. 2016. J Dairy Sci. 99. PMID: 27743673

Recently, interest in the beneficial role of probiotics in the protection and management of allergic diseases caused by immune disorders has been increasing. This study investigated the inhibitory effect of Lactobacillus plantarum L67 on induced allergic inflammatory response in bisphenol A-treated rat basophilic leukemia 2H3 (RBL-2H3) cells and mouse splenocytes. We also evaluated the applicability of L. plantarum L67 as a yogurt starter culture. We measured the ability of Lactobacillus strains to induce the production of IL-12 and IFN- γ in cultured splenocytes by ELISA. Bisphenol A (50μM)-treated RBL-2H3 cells were cotreated with a glycoprotein (18kDa) isolated from L. plantarum L67 (5-100µg/mL) for 30min. We measured the expression of mitogen-activated protein kinase (ERK and p38), AP-1 (c-Fos and c-Jun), T-bet, and GATA-binding protein 3 (GATA-3) using Western blotting to examine the differentiation of T helper cells. Furthermore, we evaluated the gene expression of IL-1β, IL-6, and IL-10 using real-time quantitative PCR. Finally, we evaluated the applicability of L. plantarum L67 as a yogurt starter by measuring pH, enumeration of bacteria, and sensory scores. Our results showed that L67 protein inhibited the phosphorylation of ERK and p38 mitogen-activated protein kinase through the transcriptional activation of AP-1 in bisphenol A-treated RBL-2H3 cells. During differentiation of T helper cells, the expression of transcription factor GATA-3 was significantly suppressed by L67 protein (100µg/mL) treatment, whereas expression of transcription factor T-bet was increased. In addition, the L67 protein significantly attenuated the expression of T helper 2-linked cytokines IL-1β, IL-6, and IL-10. These results indicate that L. plantarum L67, made available as yogurt starters and dietary supplements, has the potential to prevent allergy-related immune disorders.
Effects of Lactobacillus plantarum NCU116 on Intestine Mucosal Immunity in Immunosuppressed Mice. Junhua Xie;Qiang Yu;Shaoping Nie;Songtao Fan;Tao Xiong;Mingyong Xie. 2015. J Agric Food Chem. 63. PMID: 26651209

The effects of Lactobacillus plantarum (L. plantarum) NCU116 isolated from pickled vegetables on intestine mucosal immunity in cyclophosphamide treated mice were investigated. Animals were divided into six groups: normal group (NIM), immunosuppression group (IM), immunosuppression plus L. plantarum NCU116 groups with three different doses (NCU-H, NCU-M, and NCU-L), and plus Bifidobacterium BB12 as positive control group (BB12). Results showed that the thymus indexes of the four treatment groups were significantly higher than that of the IM group (2.02 ± 0.16) (p < 0.05) and close to the index of the NIM group (2.61 ± 0.37) at 10 days. The level of immune factor IL-2 notably increased (IM, 121 ± 9.0) (p < 0.05) and was close to 65% of NIM group's level (230 ± 10.7). The levels of other immune factors (IFN-γ, IL-10, IL-12p70, and sIgA), the gene expression levels of IL-2 and IFN-γ, and the number of IgA-secreting cells showed similar patterns (p < 0.05). However, the level of immune factor IL-4 remarkably decreased (IM, 128 ± 10.2) (p < 0.05) and was only approximately 50% of the NIM group (154 ± 18.2). The levels of other immune factors (IL-6 and IgE) and the gene expression level of IL-6 at 10 days exhibited similar changes (p < 0.05) but showed a slight recovery at 20 days, accompanied by the altered protein expression levels of T-bet and GATA-3 in the small intestine. These findings suggest that L. plantarum NCU116 enhanced the immunity of the small intestine in the immunosuppressed mice.
Antioxidant and Anti-Inflammatory Effect of Probiotic Lactobacillus plantarum KU15149 Derived from Korean Homemade Diced-Radish Kimchi. Kyoung Jun Han;Ji-Eun Lee;Na-Kyoung Lee;Hyun-Dong Paik. 2020. J Microbiol Biotechnol. 30. PMID: 32238771

Lactobacillus plantarum KU15149 was demonstrated to have probiotic behavior and functions, including antioxidant and anti-inflammatory activity. L. plantarum KU15149 obtained from homemade diced-radish kimchi has a high survival rate under artificial gastric acid (pH 2.5, 0.3% pepsin) and bile salt (0.3% oxgall) conditions. However, L. plantarum KU15149 did not produce β-glucuronidase, which is known to be a carcinogenic enzyme with resistance to several antibiotics, such as gentamycin, kanamycin, streptomycin, tetracycline, and ciprofloxacin. L. plantarum KU15149 strongly adhered to HT-29 cells and had high antioxidant activity in terms of 2,2-diphenyl- 1-picrylhydrazyl (DPPH) free radical-scavenging and β-carotene bleaching assays. L. plantarum KU15149 also exhibited a pronounced inhibition of nitric oxide (NO) production, along with expression of nitric oxide synthase (iNOS) and cyclooxygenase -2 (COX-2) as well as proinflammatory cytokines, such as TNF-α, IL-1β, and IL-6, when RAW 264.7 cells were stimulated with LPS. Therefore, L. plantarum KU15149 exhibited pharmaceutical functionality as a potential probiotic.
The Lactobacillus plantarum Eno A1 Enolase Is Involved in Immunostimulation of Caco-2 Cells and in Biofilm Development. Valeria Vastano;Annunziata Pagano;Alessandra Fusco;Gianluca Merola;Margherita Sacco;Giovanna Donnarumma. 2015. Adv Exp Med Biol. 897. PMID: 26577529

The role of probiotics in prevention and treatment of a variety of diseases is now well assessed. The presence of adhesive molecules on the cell surface of probiotics has been related to the ability to confer health benefit to the host. We have previously shown that the enolase EnoA1 of Lactobacillus plantarum, one of the most predominant species in the gut microbiota of healthy individuals, is cell surface-expressed and is involved in binding with human fibronectin and plasminogen. By means of comparative analysis between L. plantarum LM3 (wild type) and its isogenic LM3-CC1 (ΔenoA1) mutant strain, here we show that EnoA1 affects the ability of this bacterium to modulate immune response as determined by analysis of expression of immune system molecules in Caco-2 cells. Indeed, we observed induction of TLR2 expression in cells exposed to L. plantarum LM3, while no induction was detectable in cells exposed to LM3-CC1. This difference was much less consistent when expression of TLR4 was determined in cells exposed to the two strains. Pro-inflammatory (IL-6) and anti-inflammatory cytokines (IL-10, TGF-β), and the antimicrobial peptide HBD-2 were induced in Caco-2 cells exposed to L. plantarum LM3, while lower levels of induction were detected in cells exposed to LM3-CC1. We also analyzed the ability to develop biofilm of the two strains, and observed a decrease of about 65 % in the development of mature biofilm in LM3-CC1 compared to the wild type.
Evaluation of Lactic Acid Bacteria on the Inhibition of Vibrio parahaemolyticus Infection and Its Application to Food Systems. Cheng-Chih Tsai;Yung-Hsien Hung;Lan-Chun Chou. 2018. Molecules. 23. PMID: 29789490

This study tested the effect of lactic acid bacteria (LAB) inhibition on Vibrio parahaemolyticus BCRC (Bioresource Collection and Research Center) 10806 and BCRC 12865 in a food model. MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays indicated that Caco-2 cells were not damaged after a two-hour treatment with lactic acid bacteria (LAB) and V. parahaemolyticus. The LAB cell culture and supernatant effectively inhibited the growth of V. parahaemolyticus in a food model. ELISA (Enzyme-linked immunosorbent assay) results indicated the significant inhibition of TNF-α; IL-1β; and IL-6; but Lactobacillus plantarum PM 222 and L. plantarum LP 735 did not significantly affect IL-8 levels. Real-time polymerase chain reaction (PCR) results indicated that LAB could inhibit the mRNA expression of proinflammatory cytokines IL-8; IL-6; and TNF-α; which were induced by V. parahaemolyticus. After rat-received LAB; the expression levels of TNF-α; IL-6; and IL-8 in the serum decreased significantly. In intestinal histology; the rat that received L. plantarum PM 222 and L. plantarum LP 010 was able to alleviate the intestinal villi damage caused by V. parahaemolyticus; which also helped reduce cell apoptosis. In conclusion; our results indicate that LAB can inhibit inflammatory responses caused by V. parahaemolyticus and can effectively inhibit the growth of V. parahaemolyticus in food products.
The Beneficial Effects of Lactobacillus plantarum PS128 on High-Intensity, Exercise-Induced Oxidative Stress, Inflammation, and Performance in Triathletes. Wen-Ching Huang;Chen-Chan Wei;Chi-Chang Huang;Wen-Lin Chen;Hui-Yu Huang. 2019. Nutrients. 11. PMID: 30736479

A triathlon, which consists of swimming, bicycling, and running, is a high-intensity and long-term form of exercise that can cause injuries such as muscular damage, inflammation, oxidative stress, and energy imbalance. Probiotics are thought to play an important role in disease incidence, health promotion, and nutrient metabolism, but only a few studies have focused on physiological adaptations to exercise in sports science. Previous studies indicated that Lactobacillus supplementation could improve oxidative stress and inflammatory responses. We investigate the effects of Lactobacillus plantarum PS128 supplementation on triathletes for possible physiological adaptation. The triathletes were assigned to one of two groups with different exercise intensity stimulations with different time-points to investigate the effects of body compositions, inflammation, oxidative stress, performance, fatigue, and injury-related biochemical indices. L. plantarum PS128 supplementation, combined with training, can significantly alleviate oxidative stress (such as creatine kinase, Thioredoxin, and Myeloperoxidase indices) after a triathlon (p < 0.05). This effect is possibly regulated by a 6⁻13% decrease of indicated pro-inflammation (TNF-α, IL-6, and IL-8) cytokines (p < 0.05) and 55% increase of anti-inflammation (IL-10) cytokines (p < 0.05) after intensive exercise stimulation. In addition, L. plantarum PS128 can also substantially increase 24⁻69% of plasma-branched amino acids (p < 0.05) and elevate exercise performance, as compared to the placebo group (p < 0.05). In conclusion, L. plantarum PS128 may be a potential ergogenic aid for better training management, physiological adaptations to exercise, and health promotion.
Zearalenone nephrotoxicity: DNA fragmentation, apoptotic gene expression and oxidative stress protected by Lactobacillus plantarum MON03. Jalila Ben Salah-Abbès;Hela Belgacem;Khawla Ezzdini;Mosaad A Abdel-Wahhab;Samir Abbès. 2019. Toxicon. 175. PMID: 31830485

The present study was conducted to determine the abilities of the living Lactobacillus plantarum MON03 cells to degrade Zearalenone (ZEN) in liquid medium, and to elucidate the preventive effect in ZEN-contaminated balb/c mice showing kidney damage. The DNA fragmentation, Bcl-2 and Bax gene expression, caspase-3 activity, mRNA level of inflammation-regulating cytokines and histology of kidney tissues were examined. Female Balb/c mice were divided into four groups (10/group) and treated daily for 2 wk by oral gavage with lactic acid bacteria (L. plantarum MON03) 2 × 109 CFU/L, ~2 mg/kg only, ZEN (40 mg/kg BW) only, ZEN (40 mg/kg BW) + lactic acid bacteria (L. plantarum MON03, 2 × 109 CFU/L, ~2 mg/kg). Control group received vehicle. At the end of experiment, the kidney was collected for the determination of DNA fragmentation, Bcl-2 and Bax gene expression,caspase-3 activity, Malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) content, as well as for any alterations in expression of total antioxidant activity (TAC) and mRNA levels of inflammation-regulating cytokines (e.g., IL-10, IL-6, TNF-alpha). The results indicated that, kidney cells exposure to ZEN led to increased caspase-3 activity, MDA, and IL-10, IL-6, TNF-alpha and Bax mRNA levels, but decreased TAC content and down-regulated expression of GSH-Px and CAT and Bcl-2 mRNA. Co-treatment with ZEN plus LP suppressed the levels of DNA fragmentation; normalized kidney MDA and increased CAT levels, up-regulated expression of GSH-Px and CAT, and normalized mRNA levels of the analyzed cytokines. It's concluded that ZEN might have toxic effects in kidney. Further, it can be seen that use of LP induced protective effects against the oxidative stress and kidney toxicity of ZEN in part through adhesion (and so likely diminished bioavailability).
Production of nitric oxide, tumor necrosis factor-alpha and interleukin-6 by RAW264.7 macrophage cells treated with lactic acid bacteria isolated from kimchi. Haeng Jeon Hur;Ki Won Lee;Hyong Joo Lee. 2005. Biofactors. 21. PMID: 15630182

The present study investigated immunopotentiating activities of lactic acid bacteria isolated from kimchi (KLAB) in vitro. A RAW264.7 macrophage cell line was stimulated with four strains of KLAB and two strains of bifidobacteria, and then the production of nitric oxide (NO) and cytokines--tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6)--was determined. Lactobacillus plantarum, a lactic acid bacteria involved in the latent fermentation of kimchi, was the most effective for the generation of NO, TNF-alpha, and IL-6 in macrophage. All strains generated NO, while for the TNF-alpha and IL-6, only L. plantarum had significant activity. Our results indicate that L. plantarum plays an important role in the immunopotentiating activity of kimchi.
Effect of Lactobacillus plantarum CJLP243 on the growth performance and cytokine response of weaning pigs challenged with enterotoxigenic Escherichia coli. J S Lee;E G Awji;S J Lee;D D Tassew;Y B Park;K S Park;M K Kim;B Kim;S C Park. 2012. J Anim Sci. 90. PMID: 22859771

The purpose of the present study was to evaluate the effect of diets containing Lactobacillus plantarum CJLP243 on the growth and cytokine response of weaning pigs (Sus scrofa) challenged with enterotoxigenic Escherichia coli (ETEC). In a 28-d experiment (14 d before and 14 d after challenge), a total of 108 pigs at 20 ± 1 d of age were allotted to 1 of 6 diets. These were a control diet without ETEC challenge (CON) and 5 treatment diets with ETEC challenge, including a control diet with ETEC challenge (negative control, NC); a positive control diet containing antibiotics (PC); control diet plus (10(8), 10(9), or 10(10)) cfu/kg L. plantarum CJLP243 (T1, T2, and T3, respectively). After challenge, NC showed the least ADFI, whereas PC and T3 had the greatest ADFI (P = 0.002). The ADG of PC, T2, and T3 were greater (P = 0.001) than that of CON, NC, and T1 during wk 1 to wk 2. During wk 3 to wk 4, a marked decline was seen in NC (P = 0.001) compared with CON, whereas PC and T3 showed increased ADG (P = 0.001). The overall ADG of PC and T3 were greater (P < 0.001) than the remaining groups. The PC and T3 had the greatest G:F during the second 2 wk (P = 0.002), and the overall 4-wk experimental period (P = 0.003). At 3 h after challenge, all groups except CON had greater rectal temperatures (RT; P < 0.05). The RT decreased to prechallenge temperatures at 9 h (PC and T3), 24 h (T1 and T2), and remained increased until d 7 in NC. At 7 and 14 d postinfection, the number of animals detected positive for ETEC by PCR assay was the greatest in NC; however, the PC group had the fewest ETEC-positive animals (P < 0.05), which was similar to T3. All challenged pigs, except T2, had greater concentrations of serum haptoglobin compared with CON, with the greatest concentration observed in NC (P < 0.001). Challenged pigs had increased serum concentrations of tumor necrosis factor alpha (TNF-α) 3 to 48 h postinfection, with the greatest concentration of TNF-α at 48 h observed in NC (P < 0.05). Similarly, greater (P < 0.05) serum concentrations of interferon-γ were observed for 9 h (T1 and T3), 24 h (T2 and PC), and 48 h (NC) postinfection. The serum concentration of IL-6 increased (P < 0.05) for 3 h in T3 and 24 h in NC. In conclusion, our findings suggest that L. plantarum CJLP243, at a concentration of 10(10) cfu/kg, may serve as a potential alternative to antibiotic supplementation to improve the growth and health performance of weaning pigs, especially during acute inflammation of the gut after bacterial infections.
Evaluation of probiotic properties of Lactobacillus plantarum WLPL04 isolated from human breast milk. Meiling Jiang;Fen Zhang;Cuixiang Wan;Yonghua Xiong;Nagendra P Shah;Hua Wei;Xueying Tao. 2016. J Dairy Sci. 99. PMID: 26805974

Lactobacillus plantarum WLPL04, a specific strain isolated from human breast milk, was investigated for its survival capacity (acid and bile salt tolerance, survival in simulated gastrointestinal tract, inhibition of pathogens, antibiotic susceptibility, yield of exopolysaccharides) and probiotic properties (antiadhesion of pathogens, protection from harmful effect of sodium dodecyl sulfate, and antiinflammatory stress on Caco-2 cells). The results showed that Lb. plantarum WLPL04 had broad-spectrum activity against gram-positive strains (Listeria monocytogenes CMCC54007, Bacillus cereus ATCC14579, and Staphylococcus aureus CMCC26003) and gram-negative strains (Pseudomonas aeruginosa MCC10104, Shigella sonnei ATCC25931, Enterobacter sakazakii ATCC29544, Salmonella typhimurium ATCC13311, and Escherichia coli O157:H7). Antibiotic susceptibility tests showed that Lb. plantarum WLPL04 was susceptible to 8 of 14 antibiotics (e.g., erythromycin and nitrofurantoin) and resistant to 6 of 14 antibiotics (e.g., kanamycin and bacitracin). Lactobacillus plantarum WLPL04 was able to survive at pH 2.5 for 3h and at 0.45% bile salt for 12h, suggesting that it can survive well in the gastrointestinal tract. In addition, the exopolysaccharide yield of Lb. plantarum WLPL04 reached 426.73 ± 65.56 mg/L at 24h. With strategies of competition, inhibition, and displacement, Lb. plantarum WLPL04 reduced the adhesion of E. coli O157:H7 (35.51%), Sal. typhimurium ATCC 13311 (8.10%), and Staph. aureus CMCC 26003 (40.30%) on Caco-2 cells by competition, and subsequently by 59.80, 62.50, and 42.60%, respectively, for the 3 pathogens through inhibition, and by 75.23, 39.97, and 52.88%, respectively, through displacement. Lactobacillus plantarum WLPL04 attenuated the acute stress induced by sodium dodecyl sulfate on Caco-2 cells and significantly inhibited the expression of inflammatory cytokines (IL-6, IL-8 and tumor necrosis factor-α) on Caco-2 cells but increased IL-10 expression in vitro compared with the Salmonella-treated group. In summary, Lb. plantarum WLPL04 from breast milk could be considered as a probiotic candidate for dairy products to promote human health.
Chlorogenic Acid Combined with Lactobacillus plantarum 2142 Reduced LPS-Induced Intestinal Inflammation and Oxidative Stress in IPEC-J2 Cells. Orsolya Palócz;Erzsébet Pászti-Gere;Péter Gálfi;Orsolya Farkas. 2016. PLoS One. 11. PMID: 27861533

This study was carried out to investigate protective effect of chlorogenic acid against lipopolysaccharide-induced inflammation and oxidative stress in intestinal epithelial cells. As a marker of inflammatory response, IL-6, IL-8, TNF-α mRNA and protein levels, furthermore, COX-2 mRNA level were followed up. Intracellular redox status and extracellular H2O2 level were also monitored by two fluorescent assays (DCFH-DA, Amplex Red). Moreover, the effect of gut microbiota metabolites in the above mentioned processes was taken into account in our model using Lactobacillus plantarum 2142 bacterial strain. Our data revealed that chlorogenic acid had significant lowering effect on the inflammatory response. Treatment with chlorogenic acid (25-50 μM) significantly decreased gene expression and concentration of proinflammatory cytokines IL-6 and IL-8 compared to LPS-treated cells. COX-2 and TNF-α mRNA levels were also reduced. Furthermore, chlorogenic acid reduced the level of reactive oxygen species in IPEC-J2 cells. Simultaneous application of chlorogenic acid and Lactobacillus plantarum 2142 supernatant resulted protective effect against LPS-induced inflammation and oxidative stress as well.
Lactobacillus plantarum CAU1055 ameliorates inflammation in lipopolysaccharide-induced RAW264.7 cells and a dextran sulfate sodium-induced colitis animal model. Sun-Hae Choi;Sun-Hee Lee;Min Gon Kim;Hong Jin Lee;Geun-Bae Kim. 2019. J Dairy Sci. 102. PMID: 31155246

This study aimed to screen lactic acid bacteria (LAB) for their anti-inflammatory activity by using RAW264.7 cells and dextran sulfate sodium (DSS)-induced colitis. In all, 192 LAB strains were isolated from healthy human feces, of which 8 strains showed excellent nitric oxide (NO) inhibitory activity. Peptidoglycan extracts of these 8 LAB strains were subjected to NO assay, Western blot, and ELISA. Among the 8 tested strains, extracts of 4 strains significantly inhibited the production of NO, related enzyme activities such as inducible nitric oxide synthase and cyclooxygenase 2, and key cytokines such as tumor necrosis factor-α and IL-6 in RAW264.7 cells. The 4 strains belonged to Lactobacillus (CAU1054, CAU1055, CAU1064, and CAU1301). Oral administration of the 4 strains inhibited DSS-induced body weight loss, colon shortening, and colon damage in ICR mice. The colon tissue of the mice treated with Lactobacillus plantarum strain CAU1055 had significantly reduced levels of inducible nitric oxide synthase, cyclooxygenase 2, tumor necrosis factor-α, and IL-6. We found that strain CAU1055 could be used as a candidate probiotic strain for the prevention and treatment of inflammatory bowel disease. Further studies are warranted to confirm the mechanisms of interaction between peptidoglycan of L. plantarum strain CAU1055 and upstream cellular signaling mediators.
Aryl hydrocarbon receptor ligands enhance lung immunity through intestinal IKKβ pathways. Tzyy-Bin Tsay;Pei-Hsuan Chen;Lee-Wei Chen. 2019. J Transl Med. 17. PMID: 31488203

BACKGROUND: Infection by antibiotic-resistant microorganisms is common in intensive care units and has become a global problem. Here, we determined the effect of aryl hydrocarbon receptor (AhR) stimulation on antibiotics-induced systemic defense impairment and its mechanisms. METHODS: C57BL/6 wild-type (WT) mice received combined antibiotics with or without Ahr ligands (tryptophan and indole), or dead Lactobacillus plantarum supplementation. The defense mechanisms against Pseudomonas aeruginosa infection in the lung were examined. RESULTS: Antibiotic treatments decreased the phagocytic activity, physiological activity, and the peroxynitrite production of alveolar macrophage (AMs). It also enhanced P. aeruginosa pneumonia-induced bacterial counts in the lung. Tryptophan and dead L. plantarum supplementation reversed antibiotic-induced intracellular adhesion molecule (ICAM) as well as IL-6 expression, and increased P. aeruginosa pneumonia-induced bacterial counts in the lung and increased phagocytic activity and peroxynitrite production of AMs. Moreover, these treatments reversed the antibiotics-induced reduction of Ahr expression, antibacterial proteins, reactive oxygen species (ROS) production, and NF-κB DNA binding activity of the intestinal mucosa and plasma IL-6 levels. P. aeruginosa counts increased and phagocytic activity of AMs and myeloperoxidase (MPO) activity decreased in intestinal IKKβ depleted mice. Antibiotics, antibiotic with tryptophan feeding, or antibiotic with dead L. plantarum feeding treatments did not change the phagocytic activity and peroxynitrite production of AMs, plasma IL-6 levels, and the expression of Ahr of intestine in intestinal IKKβ depleted mice. CONCLUSION: Antibiotic treatment impairs lung immune defenses by decreasing Ahr expression in the intestine and peroyxnitrite production of the AMs. Ahr ligands reverses antibiotic-induced lung defense against bacterial infection through intestinal ROS production and NF-κB activation. The gut is critical in maintaining lung defense mechanism through the intestinal IKKβ pathways.
Differential Immune Response of Lactobacillus plantarum 286 Against Salmonella Typhimurium Infection in Conventional and Germ-Free Mice. Tizá Teles Santos;Roberta Maria Dos Santos Ornellas;Leonardo Borges Acurcio;Sávio Henrique Cicco Sandes;Andréa Miura da Costa;Ana Paula Trovatti Uetanabaro;Jacques Robert Nicoli;Gabriel Vinderola. None. Adv Exp Med Biol. . PMID: 32415613

We aimed at evaluating in vivo the probiotic potential of Lactobacillus plantarum 286 against Salmonella enterica serov. Typhimurium. Colonization capacity and antagonistic activity were determined in feces of gnotobiotic mice. Survival to infection, translocation, histopathology, IgA and cytokine levels (IL-10, IL-6, IFN-γ, TNF-α, TGF-β) were determined both in conventional and germ-free mice followed L. plantarum 286 administration and Salmonella infection. L. plantarum 286 colonized the intestine of gnotobiotic mice, where it produced antagonistic substances against S. Typhimurium. In conventional animals, the administration of this strain increased intestinal IgA levels and reduced the inflammatory response and the tissue damage caused by S. Typhimurium. Reduction of tissue damage in the intestine and liver of germ-free animals was also observed, however the immune response elicited was different in either model. L. plantarum 286 showed in vivo probiotic properties in both murine models. Probiotic capacity results may depend on the animal model chosen.
Innate immune responses of human neonatal cells to bacteria from the normal gastrointestinal flora. Helen Karlsson;Christina Hessle;Anna Rudin. 2002. Infect Immun. 70. PMID: 12438343

The hygiene hypothesis postulates that the prevalence of allergy has increased due to decreased microbial stimulation early in life, leading to delayed maturation of the immune system. The aim of this study was to examine the cytokine pattern produced from cord blood mononuclear cells relative to adult cells after stimulation with bacterial strains from the normal flora. Mononuclear cells from cord and adult blood samples were stimulated with the following bacteria: Bifidobacterium adolescentis, Enterococcus faecalis, Lactobacillus plantarum, Streptococcus mitis, Corynebacterium minutissimum, Clostridium perfringens, Bacteroides vulgatus, Escherichia coli, Pseudomonas aeruginosa, Veillonella parvula, and Neisseria sicca. The levels of interleukin 12 (IL-12), tumor necrosis factor alpha (TNF-alpha), IL-10, and IL-6 were measured by enzyme-linked immunosorbent assay. The TNF-alpha production was also analyzed after blocking CD14, Toll-like receptor 2 (TLR-2), and TLR-4 prior to stimulation with bacteria. The levels of IL-12 and TNF-alpha were similar in cord and adult cells. Gram-positive bacteria induced considerably higher levels of IL-12 and TNF-alpha than gram-negative bacteria in both cord and adult cells. The levels of IL-6 were significantly higher in newborns than in adults, whereas the levels of IL-10 were similar in newborns and adults. Gram-negative and gram-positive bacteria induced similar levels of IL-6 and IL-10 in cord cells. L. plantarum bound or signaled through CD14, TLR-2, and TLR-4, whereas E. coli acted mainly through CD14 and TLR-4. These results indicate that the innate immune response in newborns to commensal bacteria is strong and also suggest that different bacterial strains may have differential effects on the maturation of the immune system of infants.
Immunomodulatory effects of specific bacterial components of Lactobacillus plantarum KFCC11389P on the murine macrophage cell line RAW 264.7. H Chon;B Choi;E Lee;S Lee;G Jeong. 2009. J Appl Microbiol. 107. PMID: 19486216

AIMS: The objective of this study was to investigate the ability of specific bacterial components of Lactobacillus plantarum KFCC11389P to induce anti-inflammatory mediators in cell cultures of the murine macrophage cell line, RAW 264.7. METHODS AND RESULTS: The RAW 264.7 cells were stimulated with viable bacterial cells (VC), heat-killed (HK) cells, cell walls (CW) or ultrafiltrates of metabolic products (UF). An increase in the levels of tumour necrosis factor (TNF)-alpha was observed in VC, HK and CW, but this effect was much lower in UF. VC stimulated higher levels of interleukin (IL)-6 releases as well as nitric oxide production than HK. In contrast, UF and its separated molecule, fraction 4, were much strong IL-10 inducers. Fraction 4 (8.1 kDa), especially, inhibited the production of pro-inflammatory cytokines, IL-6 (89% decrease) and TNF-alpha (55% decrease), in lipopolysaccharide (LPS)-stimulated murine macrophages. CONCLUSIONS: The results of this study indicate that metabolic products of Lact. plantarum KFCC11389P could influence the immune-modulating activity via IL-10, and pretreatment with this specific molecule could inhibit LPS-induced release of IL-6 and TNF-alpha. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings suggest that the specific molecules of Lact. plantarum KFCC11389P may be useful for the treatment of acute inflammatory responses such as Crohn's disease or ulcerative colitis.
Modulation of anti-inflammatory response in lipopolysaccharide stimulated human THP-1 cell line and mouse model at gene expression level with indigenous putative probiotic lactobacilli. V Aparna Sudhakaran;Harsh Panwar;Ritu Chauhan;Raj Kumar Duary;Rahul Kumar Rathore;Virender Kumar Batish;Sunita Grover. 2013. Genes Nutr. 8. PMID: 23728791

The anti-inflammatory potential of eight indigenous probiotic Lactobacillus isolates was evaluated in vitro in terms of modulating the expression of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in human acute monocytic leukemia (THP-1) cells under inflammatory conditions. Amongst these, Lactobacillus plantarum Lp91 was the most potent anti-inflammatory strain as it evoked a significant (P < 0.001) down-regulation of TNF-α by -1.45-fold relative to the control in THP-1 cells. However, in terms of IL-6 expression, all the strains could up-regulate its expression considerably at different levels. Hence, based on in vitro expression of TNF-α, Lp91 was selected for in vivo study in lipopolysaccharide (LPS)-induced mouse model to look at the expression of TNF-α, IL-6, monocyte chemotactic protein-1 (MCP-1), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule (ICAM-1) and E-selectin in mouse aorta. In LPS challenged (2 h) mice group fed with Lp91 for 10 days, TNF-α, IL-6, MCP-1, VCAM-1, ICAM-1 and E-selectin expressions were significantly down-regulated by 3.10-, 10.02-, 4.22-, -3.14-, 2.28- and 5.71-fold relative to control conditions. In conclusion, Lp91 could serve as a candidate probiotic strain to explore it as a possible biotherapeutic anti-inflammatory agent against inflammatory diseases including cardiovascular disease.
Lactobacillus plantarum CLP-0611 ameliorates colitis in mice by polarizing M1 to M2-like macrophages. Se-Eun Jang;Myung Joo Han;Se-Young Kim;Dong-Hyun Kim. 2014. Int Immunopharmacol. 21. PMID: 24815859

The TNF-α expression-inhibitory effect of lactic acid bacteria (LAB) isolated from kimchi were measured in lipopolysaccharide (LPS)-stimulated peritoneal macrophages. Among the LAB evaluated, Lactobacillus plantarum CLP-0611 inhibited the IL-1β and IL-6 expression, as well as the NF-κB and AP1 activation in LPS-stimulated peritoneal macrophages. Therefore, we investigated its inhibitory effect on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. TNBS significantly induced colon shortening, as well as myeloperoxidase activity and macroscopic score. Oral administration of CLP-0611 significantly reduced TNBS-induced body weight loss, colon shortening, myeloperoxidase activity, IRAK-1 phosphorylation, NF-κB and MAP kinase (p38, ERK, JNK) activation, and iNOS and COX-2 expression. CLP-0611 also inhibited TNBS-induced expression of TNF-α, IL-1β, and IL-6. However, IL-10 expression was induced. CLP-0611 also induced the production of M2 macrophage markers (IL-10, arginase I and CD206). Based on these findings, CLP-0611 inhibits TLR-4-linked NF-κB and MAPK signaling pathways and polarizes M1 to M2-like macrophages, thus ameliorating colitis.
The effect of probiotics on serum levels of cytokine and endotoxin in peritoneal dialysis patients: a randomised, double-blind, placebo-controlled trial. I-K Wang;Y-Y Wu;Y-F Yang;I-W Ting;C-C Lin;T-H Yen;J-H Chen;C-H Wang;C-C Huang;H-C Lin. 2015. Benef Microbes. 6. PMID: 25609654

Inflammatory markers such as interleukin (IL)-6 and tumour necrosis factor-alpha (TNF-α) are elevated in dialysis patients and can predict cardiovascular events and all-cause mortality. Endotoxin is an important source and also another marker of inflammation in patients with chronic kidney disease. The aim of this study was to evaluate the impact of oral probiotics on serum levels of endotoxemia and cytokines in peritoneal dialysis (PD) patients. The decline of residual renal function, peritonitis episodes, and cardiovascular events were also recorded. From July 2011 to June 2012, a randomised, double-blind, placebo-controlled trial was conducted in PD patients. The intervention group received one capsule of probiotics containing 10(9) cfu Bifobacterium bifidum A218, 10(9) cfu Bifidobacterium catenulatum A302, 10(9) cfu Bifidobacterium longum A101, and 10(9) cfu Lactobacillus plantarum A87 daily for six months, while the placebo group received similar capsules containing maltodextrin for the same duration. Levels of serum TNF-α, interferon gamma, IL-5, IL-6, IL-10, IL-17, and endotoxin were measured before and six months after intervention. 39 patients completed the study (21 in the probiotics group and 18 in the placebo group). In patients receiving probiotics, levels of serum TNF-α, IL-5, IL-6, and endotoxin significantly decreased after six months of treatment, while levels of serum IL-10 significantly increased. In contrast, there were no significant changes in levels of serum cytokines and endotoxin in the placebo group after six months. In addition, the residual renal function was preserved in patients receiving probiotics. In conclusion, probiotics could significantly reduce the serum levels of endotoxin, pro-inflammatory cytokines (TNF-α and IL-6), IL-5, increase the serum levels of anti-inflammatory cytokine (IL-10), and preserve residual renal function in PD patients.
Lactobacillus plantarum NCIMB8826 ameliorates inflammation of colon and skin in human APOC1 transgenic mice. R Mariman;E Reefman;F Tielen;C Persoon-Deen;K van de Mark;N Worms;F Koning;L Nagelkerken. 2015. Benef Microbes. 7. PMID: 26689228

Genetic predisposition and environmental factors, including the gut microbiota, have been suggested as major factors in the development and progression of atopic dermatitis. Hyperlipidemic human APOC1(+/+) transgenic mice display many features of human atopic dermatitis, such as scaling, lichenification, excoriations, and pruritus, along with a disturbed skin barrier function. Cytokine analysis of serum shows an increase of various pro-inflammatory cytokines, including interleukin (IL)-12p40, IL-6, and IL-1α, but lower levels of interferon-γ. These mice also display aspects of colitis evident from macroscopic and histological abnormalities. Genome-wide transcriptome analysis of the intestine shows up-regulation of several genes associated with mast cells and eosinophils and this observation was confirmed by demonstrating increased numbers of IgE(+) and FcRε(+) mast cells in the colon and in the skin. Oral treatment with Lactobacillus plantarum NCIMB8826 resulted in decreased numbers of mast cells in the colon. Moreover, this L. plantarum strain ameliorated skin pathology, evident from improved skin barrier integrity, absence of skin thickening, and less excoriations. These results suggest that modulation of intestinal immune homeostasis contributes to the suppression of atopic dermatitis.
Lysate of Probiotic Lactobacillus plantarum K8 Modulate the Mucosal Inflammatory System in Dextran Sulfate Sodium-induced Colitic Rats. Young-Sook Ahn;Min Young Park;Jae-Ho Shin;Ji Yeon Kim;Oran Kwon. 2014. Korean J Food Sci Anim Resour. 34. PMID: 26761681

Inflammatory bowel disease (IBD) is caused by dysregulation of colon mucosal immunity and mucosal epithelial barrier function. Recent studies have reported that lipoteichoic acid (LTA) from Lactobacillus plantarum K8 reduces excessive production of pro-inflammatory cytokine. In this study, we investigated the preventive effects of lysate of Lb. plantarum K8 in dextran sulfate sodium (DSS)-induced colitis. Male Sprague-Dawley rats were orally pretreated with lysate of Lb. plantarum K8 (low dose or high dose) or live Lb. plantarum K8 prior to the induction of colitis using 4% DSS. Disease progression was monitored by assessment of disease activity index (DAI). Histological changes of colonic tissues were evaluated by hematoxylin and eosin (HE) staining. Tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) levels were measured using enzyme-linked immunosorbent assay (ELISA). The colon mRNA expressions of TNF-α, IL-6, and toll like receptor-2 (TLR-2) were examined by quantitative real-time-transcription polymerase chain reaction (qPCR). Lysate of Lb. plantarum K8 suppressed colon shortening, edema, mucosal damage, and the loss of DSS-induced crypts. The groups that received lysate of Lb. plantarum K8 exhibited significantly decreased levels of the pro-inflammatory cytokines TNF-α and IL-6 in the colon. Interestingly, colonic expression of toll like receptor-2 mRNA in the high-dose lysate of Lb. plantarum K8 group increased significantly. Our study demonstrates the protective effects of oral lysate of Lb. plantarum K8 administration on DSS-induced colitis via the modulation of pro-inflammatory mediators of the mucosal immune system.
Dietary supplementation with Lactobacillus plantarum dy-1 fermented barley suppresses body weight gain in high-fat diet-induced obese rats. Jiayan Zhang;Xiang Xiao;Ying Dong;Tian Xu;Fei Wu. 2016. J Sci Food Agric. 96. PMID: 27145037

BACKGROUND: Cereal fermentations have shown significant potential in improvement and design of the nutritional quality and health effects of foods and ingredients. In the present study, the effect of supplementary Lactobacillus plantarum dy-1 fermented barley (LFB) on obesity in high-fat diet (HFD)-induced obese rats was investigated. RESULTS: LFB treatment showed a lower rate of increase in body weight and percentage of body fat and a reversal of HFD-induced glucose intolerance, with ameliorated hyperinsulinemia, decreased levels of triglycerides and total cholesterol, and inhibited concentration of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α. Moreover, LFB treatment also showed the strongest inhibition of nuclear factor (NF)-kB activation and exhibited the greatest effects in blocking the degradation of the inhibitor of NF-kB and inhibiting p38 and JNK1 phosphorylation compared with HFD and raw barley treatment. CONCLUSIONS: It was clear that Lactobacillus plantarum dy-1 fermentation significantly improves the anti-obesity properties of barley. The results establish the foundation for ameliorating diet-induced obesity of product with LFB as nutritional supplements. © 2016 Society of Chemical Industry.
Salmosan, a β-galactomannan-rich product, in combination with Lactobacillus plantarum contributes to restore intestinal epithelial barrier function by modulation of cytokine production. M Teresa Brufau;Joan Campo-Sabariz;Sergi Carné;Ruth Ferrer;Raquel Martín-Venegas. 2016. J Nutr Biochem. 41. PMID: 27951516

Mannan-oligosaccharides (MOSs) are mannose-rich substrates with several intestinal health-promoting properties. The aim of this study was to investigate the potential capacity of Salmosan (S-βGM), a β-galactomannan-rich MOS product, to restore epithelial barrier function independently from its capacity to reduce bacterial invasion. In addition, the combination of S-βGM with the proven probiotic Lactobacillus plantarum (LP) was also tested. Paracellular permeability was assessed by transepithelial electrical resistance (TER) in co-cultures of Caco-2 cells and macrophages (differentiated from THP-1 cells) stimulated with LPS of Salmonella Enteritidis and in Caco-2 cell cultures stimulated with TNF-α in the absence or presence of 500 μg/ml S-βGM, LP (MOI 10) or a combination of both. In both culture models, TER was significantly reduced up to 25% by LPS or TNF-α stimulation, and the addition of S-βGM or LP alone did not modify TER, whereas the combination of both restored TER to values of nonstimulated cells. Under LPS stimulation, TNF-α production was significantly increased by 10-fold, whereas IL-10 and IL-6 levels were not modified. The combination of S-βGM and LP reduced TNF-α production to nonstimulated cell values and significantly increased IL-10 and IL-6 levels (5- and 7.5-fold, respectively). Moreover, S-βGM has the capacity to induce an increase of fivefold in LP growth. In conclusion, we have demonstrated that S-βGM in combination with LP protects epithelial barrier function by modulation of cytokine secretion, thus giving an additional value to this MOS as a potential symbiotic.
Heat-treated Lactobacillus plantarum increases the immune responses through activation of natural killer cells and macrophages on in vivo and in vitro models. Phil-Dong Moon;Jin Soo Lee;Hee-Yun Kim;Na-Ra Han;Inyeong Kang;Hyung-Min Kim;Hyun-Ja Jeong. 2019. J Med Microbiol. 68. PMID: 30724726

PURPOSE: Recently, Lactobacillus plantarum (nF1) has been reported to have immune-enhancing effects in an immunosuppressed-animal model. Natural killer (NK) cells and macrophages play important roles in the immune responses. However, immunomodulatory effects of heat-treated Lactobacillus plantarum-nF1 (hLp-nF1) on the activation of NK cells and macrophages have not been elucidated. METHODOLOGY: We assessed whether hLp-nF1 could elevate the activation of NK cells and macrophages using cyclophosphamide (CP)-induced immunosuppressed BALB/c mice and RAW 264.7 macrophages. A nitric oxide (NO) assay, enzyme-linked immunosorbent assay, Western blot analysis and NK cell activity assay were used to examine the effects of hLp-nF1 on the immune enhancement.Results/Key findings. Administration of hLp-nF1-elevated NK cell activities and serum levels of TNF-α, IL-2, and IL-12 in CP-induced immunosuppressed mice. In RAW 264.7 macrophages, treatment with hLp-nF1 increased the production of NO and expression of inducible NO synthase. Simultaneously, hLp-nF1 increased the production of TNF-α, IL-2, and IL-6 from RAW 264.7 cells. Finally, hLp-nF1 induced activation of nuclear factor-κB and phosphorylation of IκBα. CONCLUSION: We identified that hLp-nF1 has an immune-enhancing effect through the activation of NK cells and macrophages. Therefore, these findings suggest that hLp-nF1 would be helpful to enhance the immunity.
Extracellular transglycosylase and glyceraldehyde-3-phosphate dehydrogenase attributed to the anti-staphylococcal activity of Lactobacillus plantarum USM8613. Jia Sin Ong;Todd D Taylor;Chyn Boon Wong;Boon Yin Khoo;Sreenivasan Sasidharan;Sy Bing Choi;Hiroshi Ohno;Min Tze Liong. 2019. J Biotechnol. 300. PMID: 31095980

Increasing levels of antibiotic resistance in pathogens, including Staphylococcus aureus, remains a serious problem for public health, leading to the need for better alternative antimicrobial strategies. The antimicrobial proteins produced by Lactobacillus plantarum USM8613 attributed to its anti-staphylococcal activity were identified as extracellular transglycosylase and glyceraldehyde-3-phosphate dehydrogenase (GADPH), both with different mechanisms of action. Extracellular transglycosylase, which contains a LysM domain, exerts a cell wall-mediated killing mechanism, while GADPH penetrates into S. aureus cells and subsequently induces the overexpression of autolysis regulators, resulting in S. aureus autolysis. Both extracellular transglycosylase and GADPH exert anti-inflammatory effects in S. aureus-infected HaCaT cells by reducing the expression and production of TLR-2, hBDs and various pro-inflammatory cytokines (IL-1α, IL-1β, IL-6, TNF-α, and IL-8). Taken together, extracellular transglycosylase and GADPH produced by L. plantarum USM8613 could potentially be applied as an alternative therapeutic agent to treat S. aureus skin infections and promote skin health.
Short communication: Complete genome sequence of Lactobacillus plantarum J26, a probiotic strain with immunomodulatory activity. Ziwei Zhang;Chaoxin Man;Linlin Sun;Xinyan Yang;Mingyu Li;Wei Zhang;Yujun Jiang. 2019. J Dairy Sci. 102. PMID: 31548063

Lactobacillus plantarum J26, a significant probiotic isolated from Chinese traditional fermented dairy products, exerts a positive immunomodulatory effect by regulating the expression of immune-related genes. We investigated expression of the cytokines IL-1α, IL-1β, IL-6, and tumor necrosis factor-α in the intestinal tract of mice stimulated by L. plantarum J26. In vivo, these cytokines were upregulated, peaked on d 5, and then decreased to the control level, indicating that L. plantarum J26 could induce expression of the genes encoding these proinflammatory cytokines. Teichoic acids produced by L. plantarum are recognized as key immunomodulatory molecules involved in the regulation of the host immune response. To better understand the genetic basis of this immunomodulatory mechanism, we sequenced and analyzed the whole genome of L. plantarum J26. The genome of L. plantarum J26 contains a circular chromosome and 4 circular plasmids. Lactobacillus plantarum J26 was predicted to synthesize ribitol-type backbones of wall teichoic acid. Furthermore, orthologous average nucleotide identity (OrthoANI) values showed that the genome was highly similar (>98.00%) to other L. plantarum strains, especially to L. plantarum ST-III and JDM1. The genomic data of L. plantarum J26 provide a genetic basis to further elucidate its mechanism of immunoregulation and will facilitate its application in the functional dairy food industry.
Oral administration of Lactobacillus plantarum K68 ameliorates DSS-induced ulcerative colitis in BALB/c mice via the anti-inflammatory and immunomodulatory activities. Yen-Wenn Liu;Yu-Wen Su;Wei-Kee Ong;Tzu-Hao Cheng;Ying-Chieh Tsai. 2011. Int Immunopharmacol. 11. PMID: 21996541

Many different kinds of fermented food are consumed daily in Taiwan, such as stinky tofu, suan-tsai, and fu-tsai. We have previously reported the diversity of lactic acid bacteria (LAB) at different stages of fermentation in the production of suan-tsai and fu-tsai. In this study, the anti-inflammatory and immunomodulatory activities of Lactobacillus plantarum K68 (K68) isolated from fu-tsai were evaluated. K68 significantly inhibited the production of tumor necrosis factor-α (TNF-α) and prostaglandin E(2) (PGE(2)) in lipopolysaccharide (LPS)-induced murine macrophage RAW 264.7 cells and stimulated interferon-γ (IFN-γ) production in human peripheral blood mononuclear cells (hPBMCs). Additionally, orally administered K68 ameliorated dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in BALB/c mice. Both the disease activity index (DAI) and histological scores (HIS) showed that the severity of UC was significantly reduced by oral administration of K68. Furthermore, the production of pro inflammatory cytokines TNF-α, interleukin-1β (IL-1β), and interleukin-6 (IL-6) was significantly reduced in K68-administered group. Colonic mRNA expression levels of TNF-α, cyclooxygenase-2 (COX-2), forkhead box P3 (Foxp3), suppressors of cytokine signaling 3 (SOCS3), and toll like receptor 4 (TLR4), were also reduced in the K68-administered group. These results suggest that K68 exhibits anti-inflammatory and immunomodulatory activities that ameliorate DSS-induced experimental colitis.
Effect of microencapsulation of Lactobacillus plantarum 25 into alginate/chitosan/alginate microcapsules on viability and cytokine induction. Tao Jiang;You-Kyoung Kim;Bijay Singh;Sang-Kee Kang;Yun-Jaie Choi;Chong-Su Cho. 2013. J Nanosci Nanotechnol. 13. PMID: 23882756

Lactobacillus plantarum 25 (LP25) encapsulated into alginate/chitosan/alginate (ACA) microcapsules (LP25-ACA MCs) prepared by an extrusion methods were characterized to assess their efficacy in oral delivery. The particle sizes of LP25-ACA MCs were 1.11 +/- 0.32 mm. The loading content of LP25 was 1.11 x 10(7) colony forming unit (cfu)/microcapsule and encapsulation efficiency was above 98%. The viability of LP25 in ACA MCs was more than 65% in simulated gastric fluid (SGF, pH 2.0) and 75% in simulated small intestinal fluid (SIF, pH 7.2) up to 2 h. Encapsulated LP25 were completely released from LP25-ACA MCs in SIF and simulated colon fluid (SCF, pH 6.0) within 12 h and 8 h respectively. The viability of LP25 in ACA MCs till 5 weeks was above 58%, whereas viability of free LP25 stored at room temperature up to 5 weeks was zero. Besides, LP25-ACA MCs induced the secretion of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) from macrophages and dendritic cells showing the immunomodulatory effect of LP25. These findings demonstrate that the encapsulation of LP25 by ACA is a suitable strategy for oral delivery of probiotics.
Lactobacillus plantarum CECT 7315/7316 intake modulates the acute and chronic innate inflammatory response. Gemma Vilahur;Sergi López-Bernal;Sandra Camino;Guiomar Mendieta;Teresa Padró;Lina Badimon. 2014. Eur J Nutr. 54. PMID: 25408198

PURPOSE: Probiotics may confer health benefits for the host. Although Lactobacillus has demonstrated to stimulate the immune response, only a few strains have demonstrated immunomodulatory properties. The newly identified Lactobacillus plantarum strains CECT7315 and CECT7316 (LP3457) seem to boost the immune system in individuals that immune decline. We aimed to investigate whether LP3457 protects against inflammation and the mechanism behind. METHODS: LP3457 potential anti-inflammatory effects were assessed in an acute model LPS-induced inflammation in healthy rats and in a chronic model of low-grade inflammation in Zucker diabetic fatty (ZDF) rats. Wistar rats received LP3457 or placebo control for 20 days. Lipopolysaccharide (LPS; 1 mg/kg) was injected intraperitoneally at day 14, and animals were sacrificed 6 days after. Blood was collected at baseline (day 0) and consecutively at day 7, 14, 17, and 20 for haematological evaluation and assessment of anti-inflammatory/pro-inflammatory systemic markers. Myeloperoxidase activity was investigated in the ileum. ZDF rats received LP3457 or placebo control during 8 weeks, and changes in inflammasome-related transcripts were assessed in the ileum. RESULTS: LPS induced a comparable and significant leucocytosis 3 days post-injection (day 17) in both LP3457-treated and LP3457-untreated rats. However, the probiotic supplementation attenuated IL-1β, IL-6, and CRP release and increased anti-inflammatory IL-10 levels 6 days post-LPS induction (p < 0.05 vs. placebo). LP3457-supplemented animals also displayed lower intestinal myeloperoxidase activity (p < 0.05 vs. placebo). Chronic administration of LP3457 to ZDF rats resulted in a significant downregulation of the inflammasome signalling pathway (p < 0.05 vs. placebo). CONCLUSIONS: Intake of LP3457 attenuates both acute endotoxemia-induced and chronic metabolically induced inflammatory reactions and the inflammasome signalling pathway. The stabilization and regulation of the gut microbiota is an important target for reducing the impact of organ-related inflammatory reactions.
The toll-like receptor family protein RP105/MD1 complex is involved in the immunoregulatory effect of exopolysaccharides from Lactobacillus plantarum N14. Yo Murofushi;Julio Villena;Kyoko Morie;Paulraj Kanmani;Masanori Tohno;Tomoyuki Shimazu;Hisashi Aso;Yoshihito Suda;Kenji Hashiguchi;Tadao Saito;Haruki Kitazawa. 2014. Mol Immunol. 64. PMID: 25466614

The radioprotective 105 (RP105)/MD1 complex is a member of the Toll-like receptor (TLR) family. It was reported that RP105/MD1 cooperates with the lipopolysaccharide (LPS) receptor TLR4/MD2 complex and plays a crucial role in the response of immune cells to LPS. This work evaluated whether RP105, TLR4 or TLR2 were involved in the immunoregulatory capacities of Lactobacillus plantarum N14 (LP14) or its exopolysaccharides (EPS). EPS from LP14 were fractionated into neutral (NPS) and acidic (APS) EPS by anion exchange chromatography. Experiments with transfectant HEK(RP105/MD1) and HEK(TLR2) cells demonstrated that LP14 strongly activated NF-κB via RP105 and TLR2. When we studied the capacity of APS to activate NF-κB pathway in HEK(RP105/MD1) and HEK(TLR4) cells; we observed that APS strongly stimulated both transfectant cells. Our results also showed that LP14 and APS were able to decrease the production of pro-inflammatory cytokines (IL-6, IL-8 and MCP-1) in porcine intestinal epithelial (PIE) cells in response to enterotoxigenic Escherichia coli (ETEC) challenge. In order to confirm the role of TLR2, TLR4 and RP105 in the immunoregulatory effect of APS from LP14, we used small interfering RNA (siRNA) to knockdown these receptors in PIE cells. The capacity of LP14 and APS to modulate pro-inflammatory cytokine expression was significantly reduced in PIE(RP105-/-) cells. It was also shown that LP14 and APS were capable of upregulating negative regulators of the TLR signaling in PIE cells. This work describes for the first time that a Lactobacillus strain and its EPS reduce inflammation in intestinal epithelial cells in a RP105/MD1-dependend manner.
[The Immunomodulatory and Anti-allergy Effects of Lactobacilli]. Xi Shen;Ming Li;Lei Shi;Nan Ling;Miao He;Shu-yue Wang;Fang He. 2016. Sichuan Da Xue Xue Bao Yi Xue Ban. 47. PMID: 27263293

OBJECTIVE: This study was conducted to evaluate the potent immunomodulatory effects of Lactobacilli and their possible anti-allergic effects. METHODS: Lactobacillus plantarum LP45 (LP45), Lactobacillus acidophilus La28 (La28), Lactobacillus acidophilus 6091 (6091), Lactobacillus rhamnosus GG (LGG) were orally administrated to male BALB/C mice, respectively for 28 d. The immune organ index, serum Th1 cytokines [interferon-γ (IFN-γ), interleukin-12 (IL-12)] and Th2 cytokines IL-6 of the tested mice were analyzed with ELISA after intervention. Furthermore, La28, 6901 were also orally fed to ovalbumin (OVA)-sensitized male BALB/C. The serum total IgE of the tested mice was analyzed with ELISA after intervention. RESULTS: No statistical difference was found in immune organ index among the tested four strains. La28 significantly decreased serum IL-6 of the tested mice after 14 d and 28 d compared to those in control (P < 0.05). After 28 d, 6091 also significantly reduced serum IL-6 of the tested mice (P < 0.05). La28 significantly suppressed the increase of serum total IgE of the tested mice (P < 0.05). CONCLUSION: The present study indicates that the immunomodulatory effects of Lactobacilli might be strain-dependent. Among the tested strains of Lactobacilli, La28 and 6091 may have possibility to influence the Th2 immunity of host animal. La28 may also posse potent ability to alter IgE mediated allergy by the way to affect Th1/Th2 balance of host animal.
Synbiotics for Broiler Chickens-In Vitro Design and Evaluation of the Influence on Host and Selected Microbiota Populations following In Ovo Delivery. Aleksandra Dunislawska;Anna Slawinska;Katarzyna Stadnicka;Marek Bednarczyk;Piotr Gulewicz;Damian Jozefiak;Maria Siwek. 2017. PLoS One. 12. PMID: 28045927

Synbiotics are synergistic combinations of prebiotics and probiotics. In chickens, synbiotics can be delivered in ovo to expedite colonization of the gut by beneficial bacteria. We therefore aimed to design synbiotics in vitro and validate them in broiler chickens upon in ovo delivery. The probiotic components of the synbiotics were Lactobacillus salivarius and Lactobacillus plantarum. Their growth was assessed in MRS medium supplemented with different prebiotics. Based on in vitro results (hatchability and growth curve), two synbiotics were designed: S1 -Lactobacillus salivarius with galactooligosaccarides (GOS) and S2 -Lactobacillus plantarum with raffinose family oligosaccharides (RFO). These synbiotics were delivered to Cobb broiler chicken embryos on day 12 of incubation at optimized doses (105 cfu egg-1 of probiotic, 2 mg egg-1 of prebiotic). Post hatching, 2,400 roosters were reared (600 individuals group-1 divided into eight replicate pens). Microbial communities were analyzed in ileal and cecal digesta on day 21 using FISH. Gene expression analysis (IL1β, IL4, IL6, IL8, IL12, IL18, IFNβ, and IFNγ) was performed on days 7, 14, 21, and 42 for the spleen and cecal tonsils with RT-qPCR. Body weight and feed intake of the roosters did not differ by the treatments. Microbial populations of Lactobacillus spp. and Enterococcus spp. in the ileum were higher in S1 and S2 than in the control. In the cecum, the control had the highest bacterial counts. S1 caused significant up-regulation of IL6, IL18, IL1β, IFNγ, and IFNβ in the spleen on day 21 and IL1β on day 7 (P < 0.05). In cecal tonsils, S1 caused significant down-regulation of IL12, IL8, and IL1β on day 42 and IFNβ on day 14 (P < 0.05). S2 did not elicit such patterns in any tissues investigated. Thus, we demonstrate that divergent effects of synbiotics in broiler chickens were reflected in in vitro tests.
Whole Genome Analysis of Lactobacillus plantarum Strains Isolated From Kimchi and Determination of Probiotic Properties to Treat Mucosal Infections by Candida albicans and Gardnerella vaginalis. Bo Ram Beck;Gun-Seok Park;Yong Hyun Lee;Sunghoon Im;Do Yeun Jeong;Jihee Kang. 2019. Front Microbiol. 10. PMID: 30894844

Three Lactobacillus plantarum strains ATG-K2, ATG-K6, and ATG-K8 were isolated from Kimchi, a Korean traditional fermented food, and their probiotic potentials were examined. All three strains were free of antibiotic resistance, hemolysis, and biogenic amine production and therefore assumed to be safe, as supported by whole genome analyses. These strains demonstrated several basic probiotic functions including a wide range of antibacterial activity, bile salt hydrolase activity, hydrogen peroxide production, and heat resistance at 70°C for 60 s. Further studies of antimicrobial activities against Candida albicans and Gardnerella vaginalis revealed growth inhibitory effects from culture supernatants, coaggregation effects, and killing effects of the three probiotic strains, with better efficacy toward C. albicans. In vitro treatment of bacterial lysates of the probiotic strains to the RAW264.7 murine macrophage cell line resulted in innate immunity enhancement via IL-6 and TNF-α production without lipopolysaccharide (LPS) treatment and anti-inflammatory effects via significantly increased production of IL-10 when co-treated with LPS. However, the degree of probiotic effect was different for each strain as the highest TNF-α and the lowest IL-10 production by the RAW264.7 cell were observed in the K8 lysate treated group compared to the K2 and K6 lysate treated groups, which may be related to genomic differences such as chromosome size (K2: 3,034,884 bp, K6: 3,205,672 bp, K8: 3,221,272 bp), plasmid numbers (K2: 3, K6 and K8: 1), or total gene numbers (K2: 3,114, K6: 3,178, K8: 3,186). Although more correlative inspections to connect genomic information and biological functions are needed, genomic analyses of the three strains revealed distinct genomic compositions of each strain. Also, this finding suggests genome level analysis may be required to accurately identify microorganisms. Nevertheless, L. plantarum ATG-K2, ATG-K6, and ATG-K8 demonstrated their potential as probiotics for mucosal health improvement in both microbial and immunological contexts.
In vitro immunomodulatory effects of acidic exopolysaccharide produced by Lactobacillus planetarium JLAU103 on RAW264.7 macrophages. Ji Wang;Xiaobin Fang;Tong Wu;Li Fang;Chunlei Liu;Weihong Min. 2019. Int J Biol Macromol. 156. PMID: 31760015

Previous work from our research group has isolated and purified an acidic exopolysaccharide (named EPS103) from Lactobacillus plantarum JLAU103, which had strong in vitro antioxidant activity. In this study, we investigated the in vitro immunomodulatory activity of EPS103 in RAW264.7 macrophages with or without lipopolysaccharide (LPS) activation. The results showed that EPS103 enhanced the phagocytic activity of RAW264.7 macrophages and promoted the release of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) of RAW264.7 macrophages. Moreover, EPS103 reduced the excessive release of IL-6, TNF-α, prostaglandin E2 (PGE2), and NO, as well as inhibited the mRNA expression of IL-6, TNF-α, cyclooxygenase-2 (COX-2), and inducible NO synthase (iNOS) of RAW264.7 macrophages activated by LPS. In mechanistic studies, EPS103 exerted immunomodulatory activity via the nuclear factor-kappa B (NF-κB) signaling pathway. These findings suggest that EPS103 possesses potent dual immunomodulatory activities and could be further developed as new products for functional foods or medicines.
The Combined Use of Tea Polyphenols and Lactobacillus Plantarum ST8SH Bacteriocin in a Rabbit Model of Infection Following Femoral Fracture with Internal Fixation. Zhonghua Xu;Jin Li;Xindie Zhou;Jianbo Dai;Junjie Zhang;Yong Huang;Nanwei Xu. 2019. Med Sci Monit. 25. PMID: 30670678

BACKGROUND Worldwide, the increasing use of antibiotics has resulted in antimicrobial resistance, leading to studies to find alternative antimicrobial treatments. Tea polyphenols have antibacterial properties. Bacteriocins produced by probiotic lactobacilli can inhibit Gram-positive bacteria. This study used a rabbit model of infection, following femoral fracture with internal fixation, to evaluate the efficacy of the combined use of tea polyphenols and Lactobacillus plantarum ST8SH bacteriocin. MATERIAL AND METHODS Twenty-four New Zealand White rabbits underwent femoral fracture, internal fixation, and insertion of a mini-titanium implant, and were inoculated intravenously with suspensions of Staphylococcal bacteria. Four treatment groups included group A, injected with tea polyphenols and bacteriocins (N=6); group B, injected with cefradine and bacteriocins (N=6); group C, injected with tea polyphenols and cefradine (N=6); and group D (controls), injected with saline (N=6). Blood samples were collected at 1, 6, 12, 24, and 48 hours after the injection of bacteriocins. Biofilms that formed on the mini-titanium implant were studied by fluorescence microscopy. Serum levels of level of interleukin (IL)-8, IL-6, and tumor necrosis factor-α (TNF-α) were measured using an enzyme-linked immunosorbent assay (ELISA). RESULTS The combination of tea polyphenols and bacteriocins (group A) had a significant inhibitory effect on Staphylococcus aureus (P<0.05) and significant differences in serum levels of IL-8, TNF-α, and IL-6 levels in serum (P<0.05) when compared with groups, B, C, and D. CONCLUSIONS In a rabbit model of femoral fracture with internal fixation, the combined use of tea polyphenols and Lactobacillus plantarum ST8SH bacteriocin effectively controlled Staphylococcus aureus infection.
Health-promoting effects of Lactobacillus-fermented barley in weaned pigs challenged with Escherichia coli K88. B Koo;D Bustamante-García;J W Kim;C M Nyachoti. 2019. Animal. 14. PMID: 31426877

Fermented feeds are being considered as practical alternatives to antimicrobial growth promoters (AGP) supplemented in nursery pig diets. This study aimed to investigate health-promoting effects of fermented barley in weaned pigs challenged with Escherichia coli K88 +. A total of 37 piglets were weaned at 21 ± 1 day of age (6.41 ± 0.47 kg of BW) and assigned to either of the following five treatment groups: (1) unchallenged control (UCC; n = 7), (2) challenged control (CC; n = 7), (3) AGP (CC + 0.1% AGP; n = 7), (4) Ferm1 (challenged and fed homofermentative Lactobacillus plantarum (Homo)-fermented barley; n = 8) and (5) Ferm2 (challenged and fed heterofermentative L. buchneri (Hetero)-fermented barley; n = 8). The control diet included unfermented barley. Barley was fermented with either Homo or Hetero for 90 days under anaerobic conditions. On day 10, all pigs except those in UCC group were orally inoculated with E. coli K88 + (6 × 109 colony forming units/ml). The pre-planned orthogonal test was performed to compare (1) UCC and CC, (2) CC and AGP, (3) CC and Ferm1 + Ferm2, as well as (4) Ferm1 and Ferm2. Challenged control pigs showed shorter (P < 0.05) villus height (VH) in the duodenum and deeper (P < 0.05) crypt depth (CD) in the jejunum than UCC pigs. The AGP group had higher (P < 0.05) VH and lower (P < 0.05) IL-6 gene expression in the jejunum compared with CC group. Compared to CC, Ferm1 and Ferm2 had decreased (P < 0.05) CD in the duodenum, IL-6 gene expression in the jejunum and rectal temperature at 24 h post-challenge. Pigs fed fermented barley diets showed greater (P < 0.05) faecal abundance of Clostridium Cluster IV and Lactobacilli than those fed UCC diet. Ferm2-fed pigs showed lower (P < 0.05) concentrations of band cells, eosinophils and lymphocytes at 6, 24 and 48 h after challenge, respectively, and lower (P < 0.05) faecal abundance of Enterobacteriaceae 24 h after challenge than the Ferm1-fed pigs. In conclusion, the substitution of unfermented barley with fermented barley in a nursery diet showed similar results as those shown by AGP supplementation in terms of enhancing the intestinal morphology and modulating faecal microbiota composition, as well as down-regulating the pro-inflammatory cytokines; therefore, fermented barley can be a possible nutritional strategy for managing nursery pigs fed diets without in-feed AGP.
Structural features and immunological perception of the cell surface glycans of Lactobacillus plantarum: a novel rhamnose-rich polysaccharide and teichoic acids. Pilar Garcia-Vello;Garima Sharma;Immacolata Speciale;Antonio Molinaro;Sin-Hyeog Im;Cristina De Castro. None. Carbohydr Polym. 233. PMID: 32059908

The capsular material from Lactobacillus plantarum IMB19, an isolate from fermented vegetables, has been analyzed and our results demonstrate that most of the coat of this bacterium consists of glycerol- and ribitol-type teichoic acids, further decorated with other substituents (α-glucose and alanine), and of a capsular polysaccharide (CPS) with a linear nonasaccharide repeating unit, rich in rhamnose, interconnected to the next via a phosphodiester bridge. Stimulation of immune cells with the total capsular material resulted in the enhancement of immunostimulatory (IFNγ, TNF-α, IL-6 and IL-12) or immuno-regulatory (IL-10) cytokines in an in vitro splenocyte culture system. The capsular polysaccharide, and not the teichoic acids mixture, was responsible for the IFNγ production. Indeed, a significant increase of IFNγ along with other inflammatory cytokines, and a dose response in IFNγ expression with an EC50 of 3.16 μM was found for CPS, disclosing that this polysaccharide is a potent immunostimulatory molecule.
Amelioration of Alcohol Induced Gastric Ulcers Through the Administration of Lactobacillus plantarum APSulloc 331261 Isolated From Green Tea. Haryung Park;Donghyun Cho;Eunchong Huang;Ju Yeon Seo;Wan Gi Kim;Svetoslav Dimitrov Todorov;Yosep Ji;Wilhelm Heinrich Holzapfel. 2020. Front Microbiol. 11. PMID: 32256476

Gastric inflammation is an indication of gastric ulcers and possible other underlying gastric malignancies. Epidemiological studies have revealed that several Asian countries, including South Korea, suffer from a high incidence of gastric diseases derived from high levels of stress, alcoholic consumption, pyloric infection and usage of non-steroidal anti-inflammatory drugs (NSAIDs). Clinical treatments of gastric ulcers are generally limited to proton pump inhibitors that neutralize the stomach acid, and the application of antibiotics for Helicobacter pylori eradication, both of which are known to have a negative effect on the gut microbiota. The potential of probiotics for alleviating gastrointestinal diseases such as intestinal bowel syndrome and intestinal bowel disease receives increasing scientific interest. Probiotics may support the amelioration of disease-related symptoms through modulation of the gut microbiota without causing dysbiosis. In this study the potential of Lactobacillus plantarum APSulloc 331261 (GTB1TM), isolated from green tea, was investigated for alleviating gastric inflammation in an alcohol induced gastric ulcer murine model (positive control). Treatment with the test strain significantly influenced the expression of pro-inflammatory and anti-inflammatory biomarkers, interleukin 6 (IL6) and interleukin 10 (IL10), of which the former was down- and the latter up-regulated when the alcohol induced mice were treated with the test strain. This positive effect was also indicated by less severe gastric morphological changes and the histological score of the gastric tissues. A significant increase in the abundance of Akkermansia within the GTB1TM treated group compared to the positive control group also correlated with a decrease in the ratio of acetate over propionate. The increased levels of propionate in the GTB1TM group appear to result from the impact of the test strain on the microbial population and the resulting metabolic activities. Moreover, there was a significant increase in beta-diversity in the group that received GTB1TM over that of the alcohol induced control group.