Journal: bioRxiv

Article Title: Early-life microbial intervention reduces colitis risk promoted by antibiotic-induced gut dysbiosis

doi: 10.1101/2020.03.11.987412

Figure Lengend Snippet: (A) Study design using the IL-10 knock-out mouse model. CPZ group dams were treated with CPZ [(0.5 mg/ml) administration in drinking water] during pregnancy from day 14 of gestation until the end of the weaning period (3 weeks of age of offspring). (B) The number of 16S rRNA gene per gram of feces in male offspring from non-treated (NT) and CPZ-exposed dams at 3 and 7 weeks of age ( n = 5 per group). (C) Shannon diversity index in male offspring at 3 and 7 weeks of age. (D) PCoA plots of both unweighted and weighted UniFrac distances of 16S rRNA gene amplicon sequences in NT and CPZ male pups at 3 and 7 weeks of age, respectively. (E) Flow cytometric analyses of live CD4 + T cells expressing Foxp3 (Treg), T-bet (Th1) or RORγt (Th17) in spleens (SPLNs) and mesenteric lymph nodes (MLNs) of NT versus CPZ males at 7 weeks of age ( n = 4-5 per group). Data represent the percentage of live TCRβ + CD4 + cells. (F) IL-12p40 production of dendritic cells obtained from 7-week-old male NT (blue) and CPZ (red) pups with stimulation with fecal slurry ( n = 5 per group). * p < 0.05 and ** p < 0.01. Data represent mean ± SEM for (B), (C), (E), and (F). Female data are shown in the supplementary figure. See also Figure S1.

Article Snippet: DCs and naïve CD4 + T cells were harvested from SPLNs of mice at 7 weeks of age using a CD11c positive selection kit II (STEMCELL Technologies) and a mouse naïve CD4 + T cell Isolation kit (STEMCELL Technologies).

Techniques: Knock-Out, Amplification, Expressing

Journal: bioRxiv

Article Title: Early-life microbial intervention reduces colitis risk promoted by antibiotic-induced gut dysbiosis

doi: 10.1101/2020.03.11.987412

Figure Lengend Snippet: (A) Flow cytometric analyses of live CD4 + T cells expressing IFNγ and IL-17A stimulated via PMA and ionomycin. Cells in spleens (SPLNs) and mesenteric lymph nodes (MLNs) were obtained from NT and CPZ males at 7 weeks of age. Data represent the percentage of live TCRβ + CD4 + cells ( n = 5 per group). (B) IFNγ and IL-17A production of CD4 + T cells obtained from SPLN and MLN of 7-week-old NT and CPZ male pups with co-stimulation via anti-CD3 and anti-CD28 antibodies ( n = 5 per group). (C) The number of 16S rRNA gene per gram of feces of male recipients before CD4 + T cell transfer (NT recipients: n = 12, CPZ recipients: n = 12, Control group: n = 9). (D) Shannon diversity index of male recipients before CD4 + T cell transfer. (E) PCoA plots of both unweighted and weighted UniFrac distances of 16S rRNA gene amplicon sequences in male recipients before CD4 + T cell transfer. (F) Percent weight change (expressed as % of starting weight) of male recipients after CD4 + T cell transfer. (G) Survival rate of male recipients during the 8-week observation period. (H) Fecal lipocalin-2 (LCN-2) levels before CD4 + T cell transfer (week 0) and at 2 weeks after post-transfer (week 2) in male recipients. (I) Histological assessment of colons from the surviving male recipients at week 8 (NT group: n = 10, and CPZ group: n = 9). Representative H&E histological sections of colon are presented for NT recipients, CPZ recipients, and the control group. * p < 0.05, ** p < 0.01, and *** p < 0.001. Data represent mean ± SEM for (A)-(D), (F), (H), and (I). Female data are shown in the supplementary figure. See also Figure S2.

Article Snippet: DCs and naïve CD4 + T cells were harvested from SPLNs of mice at 7 weeks of age using a CD11c positive selection kit II (STEMCELL Technologies) and a mouse naïve CD4 + T cell Isolation kit (STEMCELL Technologies).

Techniques: Expressing, Amplification