5mRNA transcripts (Fig

5mRNA transcripts (Fig. of this cytokine (Fig. 1in WT mice on day 0, at 3, 6, and 24 h p.c., and on days 2, 5, 8, 15, and 23 p.c., normalized to -actin. Relative expression (RQ) was calibrated to 0 p.c. salivary glands. ** 0.01; *** 0.001 versus day 0. Results represent two or three experiments with four glands analyzed per group. ( 0.05; GEE analysis followed by Sidaks significance test. TLO Formation Is Rabbit Polyclonal to DAPK3 usually Impaired in the Absence of IL-22. To evaluate the consequence of IL-22 deficiency in TLO formation and autoantibody production, we moved our analysis to mice. First, resting salivary glands from and WT mice were TGR-1202 evaluated for the presence of potential anatomical or structural differences that could interfere with computer virus infectivity. No differences were found between WT and mice in resting condition by histological examination and flow cytometry analysis of the salivary gland epithelial component (Fig. S2 mice (Fig. S2mice develop significantly smaller salivary gland lymphocytic aggregates than WT mice (Fig. 2mice were characterized by a defect in B-cell accumulation (shown as a decreased B-cell/T-cell ratio) and follicular business TGR-1202 (Fig. 2 and transcripts (Fig. 2expression was maintained post immunization (Fig. S3mice, quantitative PCR was performed on cannulated WT and mice. Preserved IL-17 up-regulation was observed in mice p.c., suggesting that the effect on TLO formation in the Il-22?/? mice is not dependent on IL-17 (Fig. S3mice at days 8 and 15 p.c. ((white bars) mice. Data present the mean SD from two different experiments with two or three mice (four or six salivary glands) per group; ** 0.01. ((white bars) mice. Data present the mean SD from two different experiments (four or six salivary glands); ** 0.01. (gene (normalized to -actin), in week 3 p.c. salivary glands from WT and mice. RQ values are relative to day 0 p.c. mRNA; data represent the mean SD of three impartial experiments with four to six salivary glands analyzed in each experiment. *** 0.001. (mice at week 3 p.c. (nuclear staining in gray, autoantibody reactivity in green) (serum dilution 1:80). ANA, antinuclear antibody. (mice at day 23 p.c. Data present the TGR-1202 mean SD of two experiments with five cannulated mice; * 0.05; unpaired test. Open in a separate windows Fig. S2. (mice at day 0 p.c. (((red circles) mice calculated at days 5, 8, 15, and 23 p.c. Open in a separate windows Fig. S3. (mRNA obtained from the spleens of nonimmunized (day 0) and immunized (day 8) (black symbols) and WT (white symbols) mice. Results are presented as CT value. ((white bars) mice at 3 h p.c. and days 2, 5, 8, 15, and 23 p.c. Transcripts were normalized to -actin. The relative expression values were calibrated to salivary gland values on day 0 p.c. Data present the mean and SD of two experiments with four or six mice analyzed per group. GEE analysis was followed by Sidaks significance test. mice. Indeed, total mRNA transcript and protein expression for CXCL13 were significantly decreased in mice compared with WT mice (Fig. 3and Fig. S4). These data were confirmed on sorted gp38+ stromal cells that showed a significant decrease in the transcript for CXCL13 in mice (Fig. 3mice in CXCL13 expression on sorted epithelial cells or gp38? stromal cells (Fig. 3mice (Fig. 3mice that showed decreased expression of the lymphoid chemokines CXCL13 and CXCL12 p.c., phenocopying the observation in mice (Fig. S4 and mRNA in FACS-sorted CD45?EpCAM?CD31?gp38+ cells (black bars) in comparison.