IL-10-secreting Tregs were reported to suppress the generation of Th2 immunity and IgE production. nuclear factor kappa B p65. Results OVA-sensitised mice showed mitigation of respiratory manifestations, alleviation of lung inflammation and congestion, and the presence GSK-J4 of an intact intestinal villus structure. Furthermore, OVA-specific immunoglobulin E (IgE), OVA-specific-IgG1, and plasma histamine levels were declined in mice treated with JC7 than in OVA-sensitised mice. In addition, interferon- (IFN-) and interleukin 10 (IL-10) levels were significantly increased, while IL-4 and IL-17A levels were clearly decreased in mice that had undergone oral administration of JC7, compared with OVA-sensitised mice. These findings Rabbit Polyclonal to MRPS21 indicated imbalances of T helper cell type 1 (Th1)/Th2 and regulatory T cells (Treg)/Th17, which were confirmed by quantitative polymerase chain reaction (PCR). Western blotting demonstrated that this expression levels of phosphorylated IB and nuclear factor kappa B p65 were significantly increased in OVA-sensitised mice, but these changes were partly reversed after treatment GSK-J4 with JC7. Oral administration of JC7 increased the richness, diversity, and evenness of cecum microbiota, characterised by higher Bacteroidetes abundance and lower Firmicutes abundance. Additionally, the intestinal microbial community composition was significantly altered in the OVA-sensitised group, indicating a disordered intestinal microbiota that was restored by the oral administration of JC7. Conclusion Overall, JC7 can prevent food allergy by rectifying Th1/Th2 and Treg/Th17 imbalances, combined with modifications of disordered intestinal microbiota. Keywords: Food allergy, Th1/Th2 imbalance, Treg/Th17 imbalance, JC7 exhibited immunomodulatory effects in our previous in vitro study. In the present study, an ovalbumin (OVA)-induced food allergy animal model was established using BALB/c mice; JC7 was orally administered to mice after treatment with OVA. Serum levels of OVA-specific immunoglobulins and various cytokines were tested, and the cecum microbiota was analysed to explore the associations between these indicators and OVA-induced food allergy. Materials and methods Bacterial strains and culture conditions JC7 was isolated from naturally fermented pickles purchased from the Yanbian area of GSK-J4 Jilin Province. JC7 was cultured at 37?C for 16C18?h. Cell pellets were washed twice and adjusted to 1 1.0??109 colony-forming units/mL. They were then stored at 4?C until GSK-J4 oral administration to mice. Animals and experimental design Six-week-old female BALB/c mice were housed in a controlled environment and had free access to standard feed and sterilised tap water. Mice were randomly divided into three groups (JC7 (200?L/mouse with concentration of 1 1.0??109?CFU/mL) by oral gavage three times per week from day 0 for 3 consecutive weeks; they underwent intraperitoneal injection of 20?g OVA (Sigma, St. Louis, MO, USA) made up of Complete Freunds adjuvant (1:1, v/v) on days 0, 7, 14, and 21. Mice from OVA group were sensitised to OVA made up of Complete Freunds adjuvant once per week for 3?weeks by intraperitoneal injection, simultaneously they received saline by oral gavage three times per week for 3?weeks. In parallel, the control group received 200?L saline (0.9%) by oral gavage three times per week, followed by intraperitoneal injection of saline. From day 21 to day 28, the mice were orally challenged twice by OVA (50?mg/mL). See Fig.?1 for the experimental schedule. On day 27, all mice were fasted overnight and on day 28, the blood samples were collected by removing the eyeball and mice were killed by dislocation 30?min after the second oral challenge. Open in a separate windows Fig. 1 Experimental schedule. Six-week-old female BALB/c mice were randomly divided into three groups (JC7 (200?L/mouse GSK-J4 with concentration of 1 1.0??109?CFU/mL) by oral gavage three times per week. In parallel, they underwent intraperitoneal injection of 20?g OVA containing Complete Freunds adjuvant. Mice from OVA group were sensitised to OVA once per week by intraperitoneal injection, simultaneously they received saline by oral gavage. The control group received 200?L saline (0.9%) by oral gavage, followed by intraperitoneal injection of saline. From day 21 to day 28, the mice were orally challenged twice by OVA (50?mg/mL) Culture of spleen cells After killing by dislocation, the mice were soaked in 75% alcohol for 5?min and then under sterile environment, the spleens were removed from BALB/c mice and washed repeatedly by 1?mL syringe until they were pale red in colour using RPMI-1640 medium supplemented with 100 U/mL penicillin, 100?g/mL streptomycin, and 10% foetal bovine serum (HyClone Laboratories, Inc., Omaha, NE, USA). Erythrocytes were lysed using lysis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China). The cells were adjusted to a final concentration of 2.5??106 cells/mL and cultured in 6-well tissue culture plates in a total of 2.5?mL medium containing OVA (5?g/mL) at 37?C for 48?h in the presence or.