Lactobacillus rhamnosus HDB1258 modulates gut microbiota-mediated immune response in mice with or without lipopolysaccharide-induced systemic inflammation.

BACKGROUND: Gut microbiota closely communicate in the immune system to maintain a balanced immune homeostasis in the gastrointestinal tract of the host. Oral administration of probiotics modulates gut microbiota composition. In the present study, we isolated Lactobacillus rhamnosus HDB1258, which induced tumor necrosis factor (TNF)-α and interleukin (IL)-10 expression in macrophages, from the feces of breastfeeding infants and examined how HDB1258 could regulate the homeostatic immune response in mice with or without lipopolysaccharide (LPS)-induced systemic inflammation. RESULTS: Oral administration of HDB1258 significantly increased splenic NK cell cytotoxicity, peritoneal macrophage phagocytosis, splenic and colonic TNF-α expression, TNF-α to IL-10 expression ratio, and fecal IgA level in control mice, while Th1 and Treg cell differentiation was not affected in the spleen. However, HDB1258 treatment significantly suppressed peritoneal macrophage phagocytosis and blood prostaglandin E2 level in mice with LPS-induced systemic inflammation. Its treatment increased LPS-suppressed ratios of Treg to Th1 cell population, Foxp3 to T-bet expression, and IL-10 to TNF-α expression. Oral administration of HDB1258 significantly decreased LPS-induced colon shortening, myeloperoxidase activity and NF-κB+/CD11c+ cell population in the colon, while the ratio of IL-10 to TNF-α expression increased. Moreover, HDB1258 treatment shifted gut microbiota composition in mice with and without LPS-induced systemic inflammation: it increased the Cyanobacteria and PAC000664_g (belonging to Bacteroidetes) populations and reduced Deferribacteres and EU622763_s group (belonging to Bacteroidetes) populations. In particular, PAC001066_g and PAC001072_s populations were negatively correlated with the ratio of IL-10 to TNF-α expression in the colon, while the PAC001070_s group population was positively correlated. CONCLUSIONS: Oral administered HDB1258 may enhance the immune response by activating innate immunity including to macrophage phagocytosis and NK cell cytotoxicity in the healthy host and suppress systemic inflammation in the host with inflammation by the modulation of gut microbiota and IL-10 to TNF-α expression ratio in immune cells.

Antioxidant and immunostimulatory effect of potential probiotic Lactobacillus paraplantarum SC61 isolated from Korean traditional fermented food, jangajji.

This study aimed to isolate and demonstrate their antioxidant and immunostimulatory activities of potential probiotics. The isolated strains, S. Pum19, SC28, and SC61 showed potential probiotic properties including stability in artificial gastric and bile conditions, non-production of ß-glucuronidase, suitable antibiotic susceptibility, and attachment to intestinal cells. S. Pum19, SC28, and SC61 strains were identified as Leuconostoc citreum, Pediococcus pentosaceus, and Lactobacillus paraplantarum, respectively. Of the 3 potential probiotic LAB strains, intact cells of L. paraplantarum SC61 showed higher antioxidant activity, including DPPH radical scavenging, ß-carotene bleaching inhibition, reducing power, superoxide anion scavenging, and ABTS radical scavenging activity. In addition, L. paraplantarum SC61 produced the most nitric oxide production and its mRNA expression level for iNOS, IL-1ß, IL-6, and TNF-α were superior to those of L. rhamnosus GG. Therefore, L. paraplantarum SC61 was demonstrated to exhibit antioxidant and immunostimulatory activity and to have potential use as a probiotic product.

In vitro characterization of Lactobacillus brevis KU15006, an isolate from kimchi, reveals anti-adhesion activity against foodborne pathogens and antidiabetic properties.

This study aimed at evaluating the functional and probiotic properties of three lactic acid bacterial (LAB) strains isolated from kimchi. The selected LAB strains, which had potential probiotic functions, were identified by 16S rRNA sequence analysis as Lactobacillus brevis G1, L. brevis KU15006, and Lactobacillus curvatus KCCM 200173. All LAB strains were able to tolerate incubation at pH 2.5 with 0.3% pepsin for 3 h and with 0.3% Oxgall for 24 h and showed similar enzyme production levels, antimicrobial activities, and antibiotic susceptibilities. L. brevis G1 and KU15006 presented higher adhesion ability, auto-aggregation, and cell surface hydrophobicity than Lactobacillus rhamnosus KCTC 12202BP, a commercial strain used as positive control. All LAB strains showed 50-60% co-aggregation activity with selected foodborne pathogens. L. brevis KU15006 showed anti-adhesion activity against Escherichia coli and Salmonella Typhimurium. In addition, cell-free supernatant and cell-free extract from L. brevis KU15006 displayed the highest inhibitory activities against α-glucosidase. These results indicate that L. brevis KU15006 has the best properties, with pathogen antagonistic and antidiabetic activity, for use in probiotic products.

TNF-α differentially modulates subunit levels of respiratory electron transport complexes of ER/PR +ve/-ve breast cancer cells to regulate mitochondrial complex activity and tumorigenic potential.

BACKGROUND: Tumor necrosis factor-α (TNF-α) is an immunostimulatory cytokine that is consistently high in the breast tumor microenvironment (TME); however, its differential role in mitochondrial functions and cell survival in ER/PR +ve and ER/PR -ve breast cancer cells is not well understood. METHODS: In the current study, we investigated TNF-α modulated mitochondrial proteome using high-resolution mass spectrometry and identified the differentially expressed proteins in two different breast cancer cell lines, ER/PR positive cell line; luminal, MCF-7 and ER/PR negative cell line; basal-like, MDA-MB-231 and explored its implication in regulating the tumorigenic potential of breast cancer cells. We also compared the activity of mitochondrial complexes, ATP, and ROS levels between MCF-7 and MDA-MB-231 in the presence of TNF-α. We used Tumor Immune Estimation Resource (TIMER) webserver to analyze the correlation between TNF-α and mitochondrial proteins in basal and luminal breast cancer patients. Kaplan-Meier method was used to analyze the correlation between mitochondrial protein expression and survival of breast cancer patients. RESULTS: The proteome analysis revealed that TNF-α differentially altered the level of critical proteins of mitochondrial respiratory chain complexes both in MCF-7 and MDA-MB-231, which correlated with differential assembly and activity of mitochondrial ETC complexes. The inhibition of the glycolytic pathway in the presence of TNF-α showed that glycolysis is indispensable for the proliferation and clonogenic ability of MDA-MB-231 cells (ER/PR -ve) as compared to MCF-7 cells (ER/PR +ve). The TIMER database showed a negative correlation between the expressions of TNF-α and key regulators of mitochondrial OXPHOS complexes in basal breast vs lobular carcinoma. Conversely, patient survival analysis showed an improved relapse-free survival with increased expression of identified proteins of ETC complexes and survival of the breast cancer patients. CONCLUSION: The evidence presented in our study convincingly demonstrates that TNF-α regulates the survival and proliferation of aggressive tumor cells by modulating the levels of critical assembly factors and subunits involved in mitochondrial respiratory chain supercomplexes organization and function. This favors the rewiring of mitochondrial metabolism towards anaplerosis to support the survival and proliferation of breast cancer cells. Collectively, the results strongly suggest that TNF-α differentially regulates metabolic adaptation in ER/PR +ve (MCF-7) and ER/PR -ve (MDA-MB-231) cells by modulating the mitochondrial supercomplex assembly and activity.

Gut Microbiota-Mediated Immunomodulatory Effects of Lactobacillus rhamnosus HDB1258 Cultured in the Lava Seawater in the Colitis Mouse Model.

The immunomodulatory effects of Lactobacillus rhamnosus HDB1258 were evaluated in mice with colitis induced by Klebsiella oxytoca (KO). L. rhamnosus HDB1258 was cultured in the lava seawater (LS) to improve its probiotic properties. It increased adhesive ability to mucin with mRNA expression levels of chaperone proteins (such as GroEL/ES, DnaKJ, and HtrA). In the in vivo experiments, administration of KO caused an inflammation on the colon with gut dysbiosis. LH group (oral gavage of HDB1258 1.0 × 109 colony forming units/day) showed that inflammatory biomarkers, including IL-1ß, TNF-α, IL-6, and PGE2, were significantly decreased to less than half of the KO group, and Th1 cells were decreased in the spleen, but Treg cells were not affected. In contrast, the expression levels of secretory IgA and IL-10 were significantly increased, and the composition of gut microbiota in the LH group tended to recover similar to normal mice without any effect on the α-diversity. In conclusion, L. rhamnosus HDB1258 cultured in the LS could regulate competitively pathogenic bacteria in imbalanced flora with its improved mucin adhesive ability and was an effective immunomodulatory adjuvant for treating colitis by its regulatory function on intestinal inflammation.

Antioxidant and immune-enhancing effects of probiotic Lactobacillus plantarum 200655 isolated from kimchi.

Probiotic properties including antioxidant and immune-enhancing effects of Lactobacillus plantarum 200655 isolated from kimchi were evaluated. The tolerance of three strains (L. plantarum 200655, L. plantarum KCTC 3108, and L. rhamnosus GG to bile salts (0.3% oxgall, 24 h) was similar, and L. plantarum 200655 showed the highest tolerance to gastric juice (0.3% pepsin, 3 h). All strains presented similar autoaggregation ability. L. plantarum 200655 showed higher cell surface hydrophobicity and adhesion ability on HT-29 cells. L. plantarum 200655 did not produce ß-glucuronidase and was sensitive to ampicillin, tetracycline, chloramphenicol, and doxycycline. Additionally, L. plantarum 200655 showed the highest antioxidant effects in DPPH and ABTS radical scavenging, and ß-carotene bleaching assays. RAW 264.7 cells treated with L. plantarum 200655 produced more nitric oxide, induced nitric oxide synthase, and cytokine related to immune-enhancing effects such as interleukin-1ß and interleukin-6. Therefore, L. plantarum 200655 could be useful as a probiotic strain for older people.

Probiotic lactic acid bacteria isolated from traditional Korean fermented foods based on ß-glucosidase activity.

This study was designed to isolate lactic acid bacteria (LAB) with ß-glucosidase activity and probiotic properties from Korean fermented foods. Among nine isolates, four LAB strains had excellent survival rates at pH 2.5 with 0.3% (w/v) pepsin for 3 h and 0.3% (w/v) oxgall for 24 h. Four LAB strains did not produce ß-glucuronidase and showed adhesion ability to HT-29 cells that was superior to that shown by the reference strain Lactobacillus rhamnosus GG. All four strains were sensitive to ampicillin, tetracycline, chloramphenicol, and doxycycline. These strains were identified as Leuconostoc mesenteroides H40, Lactobacillus plantarum FI10604, L. brevis FI10700, and L. perolens FI10842 by 16S rRNA gene sequence, respectively. It was found that L. perolens FI10842 produced the highest ß-glucosidase activity (49.10 mU/mL). These results indicate that the four LAB strains could be used as potential probiotic. Especially L. perolens FI10842 could be used as a starter culture for fermentation.

Probiotic characterization of Bacillus subtilis P223 isolated from kimchi.

Probiotic characteristics of Bacillus subtilis P223 isolated from kimchi were investigated in this study. Spore cells of B. subtilis P223 showed high tolerance to artificial gastric juice (pH 2.5, 0.3% pepsin, 3 h) and bile salts (0.3% oxgall, 24 h). Spore cells of B. subtilis P223 showed more adherence to intestinal cells (HT-29 cells) than vegetative cells. In addition, B. subtilis P223 showed high autoaggregation ability, similar to a commercial strain (Bacillus clausii ATCC 700160). Moreover, its coaggregation abilities with pathogens were strong. The adherence of three pathogens (Salmonella enteritidis ATCC 13076, Listeria monocytogenes ATCC 15313, and Escherichia coli ATCC 25922) to HT-29 cells was inhibited by B. subtilis P223. It was found that B. subtilis P223 could not produce ß-glucuronidase, a carcinogenic enzyme. However, it had amylase and protease activities. Antibiotic susceptibility was measured using disk diffusion assay. It was revealed that B. subtilis P223 was only resistant to streptomycin among eight kinds of antibiotics. In addition, B. subtilis P223 showed no hemolysis activity. It did not have enterotoxin genes. Results of this study suggest that B. subtilis P223 isolated from kimchi has potential as a probiotic strain.