[PMC free article] [PubMed] [Google Scholar] 48

[PMC free article] [PubMed] [Google Scholar] 48. tKO and WT NOD mice, but IL-17 manifestation was improved under inducible Treg skewing conditions in T cells from Smad4 tKO NOD mice. Our results demonstrate that disruption of the Smad4 pathway in T cells of NOD mice raises Teff cell activation resulting in upregulation of Th17 cells, indicating that Smad4 in T cells has INCB 3284 dimesylate a protecting role in the development of SS in NOD mice. = 36; WT, = 56; male Smad4 tKO, = 71; WT, = 79). Ideals are means SD, < 0.05, compared with the WT group; sign legend as for E. E. Cumulative incidence of SS onset (combined score for both eyes over 4.0). F. Sections of lacrimal and salivary glands from 12-week aged mice were stained with hematoxylin and eosin. The dacryoadenitis and sialadenitis was obtained for focal swelling as explained in Materials and Methods. (G) Tear and saliva quantities and (H) auto-antibodies against SSA/Ro and SSB/La in sera from 12-week-old mice. (G and H) Each circle represents an individual mouse (= 7-11/group). Ideals are means SD, *< 0.05, **< 0.01. I. NIH 3T3 cells were incubated with sera from 12-week-old mice and stained with anti-mouse IgG-FITC antibody and DAPI. Scale pub = 50 m. Pathogenic markers of SS are improved in Smad4 tKO NOD mice One of the key features of SS is definitely lymphocytic infiltration of exocrine cells, such as the lacrimal glands (dacryoadenitis) and salivary glands (sialadenitis). At 12 weeks of age, severe lymphocytic infiltration INCB 3284 dimesylate was observed in the lacrimal and salivary glands of Smad4 tKO NOD mice and this became more severe at 20 weeks of age, whereas relatively less infiltration was observed in these glands of WT NOD mice (Number ?(Number1F1F and Supplementary Number 2A and 2B). We measured tear and saliva production by pilocarpine activation at 12 weeks and 20 weeks of age. Tear and saliva quantities Rabbit polyclonal to HOMER1 were significantly decreased in Smad4 tKO compared to WT NOD mice in 12 week-old mice (Number ?(Number1G).1G). At 20 weeks of age, saliva volume from Smad4 tKO NOD mice was further decreased and significantly lower than that of WT NOD mice, similar to the results of 12-week-old mice (Supplementary Number 2C). However, tear volume was not different between Smad4 tKO and WT NOD mice at 20 weeks of age (Supplementary Number 2C). These findings show that T cell-specific Smad4 deficiency resulted in an earlier functional impairment of the lacrimal and salivary glands as compared with WT NOD mice. Another key feature of SS is the presence of circulating autoantibodies, specifically anti-SSA/Ro and anti-SSB/La. Smad4 tKO NOD mice produced significantly higher levels of anti-SSA/Ro and anti-SSB/La antibodies compared with WT NOD mice (Number ?(Number1H).1H). Consistent with this, IgG anti-nuclear antibodies were also improved in sera INCB 3284 dimesylate from Smad4 tKO NOD mice compared with WT NOD mice (Number ?(Figure1I1We). We then examined the mRNA manifestation of cytokines and related transcription factors in the lacrimal and salivary glands by qRT-PCR. The manifestation of cytokines such as IFN-, IL-4, and IL-17 and these cytokine-specific transcription factors, such as T-bet for IFN-, Gata3 for IL-4 and signal transducer and activator of transcription (Stat)3 for IL-17, was significantly improved in both lacrimal and salivary glands from Smad4 tKO NOD mice compared with WT NOD mice (Number ?(Number2A2A and ?and2B).2B). When we examined the protein production of IFN- and IL-17 in the lysates of.

Scroll to top