Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. Figure S1. Numbers and frequencies of B cells
and T cells in IL-10 deficient mice. Six-week-old IL-10FL/FL Cre- mice (white bars), IL-10FL/FL CD4-Cre+ mice (black bars), and IL-10FL/FL CD19- Cre+ mice were naturally infected with L. sigmodontis. Spleen cells were stained for CD4, CD19, CD8, Foxp3, DX5, and CD3 at day 60 p.i., and cellular composition of spleen cells was analysed by flow cytometry. Representative sets of blots are shown to identify (A) CD19+ B cells, (B) CD4+CD8- T helper cells, CD8+CD4- CTL, and CD4+Foxp3+ regulatory T cells, and (C) DX5+CD3- NK cells and DX5+CD3+ NKT cells in the lymphocyte gate. Shown are the mean numbers and frequencies of cell subtypes of 4 independent. Results are Barasertib purchase expressed as mean + SEM of n ≥ 9 as total number of mice per group across experiments. *p < 0.05, **p<0.01, ***p< 0.001 between means of CD4- and CD19-specific IL-10 deficient mice compared with the IL-10FL/FL Crecontrol group, ANOVA learn more with Bonferroni posttest. Figure S2. Humoral response of L. sigmodontis-infected mice with T-cell- and B-cell-specific IL-10 deficiency. L. sigmodontis-specific Ig (IgG1, IgG2b, IgM,
and IgE) was quantified in sera from L. sigmodontis-infected IL- 10FL/FL Cre- (○ with dotted line), IL-10FL/FL CD4-Cre+ (▪ with black line), and IL-10FL/FL CD19-Cre+ mice (♦ with grey line) at indicated time points of infection (d0 to d60 p.i.) by ELISA. Results are expressed as arbitrary Carbachol units (a.u.) (OD450 of samples subtracted by OD450 of blank). Serum dilutions were 1:1000 for IgG1 and IgM, and 1:100 for IgG2b
and IgE. Graphs show combined results of 2 independent experiments (n ≥ 9). Error bars show SEM. “
“The aim of this study was to evaluate the immunomodulatory properties of Enterococcus faecium JWS 833 (JWS 833) isolated from duck intestine and compare them to those of Lactobacillus rhamnosus GG (LGG), a proven immunity-enhancing probiotic. To investigate the immune-enhancing properties of JWS 833, production of nitric oxide (NO) and cytokines was measured in mouse peritoneal macrophages. In addition, a Listeria monocytogenes challenge model was used in the assessment. It was found that heat-killed JWS 833 stimulates mouse peritoneal macrophages to produce NO, interleukin-1 β (IL-1β) and tumor necrosis factor-α (TNF-α) and that oral administration of viable JWS833 enhances NO, IL-1β and TNF-α synthesis upon L. monocytogenes challenge. Moreover, mice fed with JWS 833 were partially protected against lethal challenge with L. monocytogenes. JWS 833 strain has significantly greater immunostimulatory properties than LGG. Moreover, JWS 833 strain partially protects mice against lethal challenge with L. monocytogenes. JWS 833, a novel strain of E. faecium isolated from duck intestine, is potentially a useful feed supplement for controlling pathogens and enhancing host immune responses.