Effect of IL-17A, IL-17F or IL-22 on the distribution of junction proteins in ARPE-19 monolayer.

<p>Cells were incubated with or without 50 ng/ml IL-17A, IL-17F or IL-22 for 6 days, then fixed and immunolabeled with ZO-1 or occludin. Immunostaining for ZO-1 and occludin in untreated or IL-22-treated ARPE-19 monolayer showed a continuous labeling in the region of cell-cell contact. Incubation with IL-17A or IL-17F caused a marked disruption of ZO-1 and occludin staining. The immunostaining shown is representative of three independent experiments. Scale bar  = 15 µm.</p

Effect of IL-17A, IL-17F or IL-22 on transepithelial diffusion rate of FITC-dextran in ARPE-19 monolayer.

<p>Stimulation of ARPE-19 monolayer with 50 ng/ml IL-17A, IL-17F or IL-22 for 6 days induced a higher FITC-dextran diffusion rate at 24 hours compared with the control group. A diffusion percentage approaching 100% indicates that the amount of dextran-FITC in the upper and lower chamber approaches the same values. IL-22 had no effect on diffusion rate. Data were shown as the means±SEM of four independent experiments. *<i>p</i><0.05 versus the control group.</p

IL-17A and IL-17F but not IL-22 promoted chemokines and IL-6 production in ARPE-19 cells.

<p>Confluent ARPE-19 cells were stimulated with different concentrations of IL-17A, IL-17F or IL-22 as indicated. After 24 hours of incubation, protein concentrations of CXCL8, CCL2, CCL20 and IL-6 released into the supernatants were measured by ELISA. Data were shown as the means±SEM of four independent experiments. **<i>p</i><0.01 versus the control group.</p

Expression of IL-22R and IL-17RC in ARPE-19 cells.

<p>(A) Coexpression of IL-22R and IL-17RC in ARPE-19 cells was shown by immunocytochemistry. Scale bar  = 75 µm. (B) Western blot revealed that ARPE-19 cells expressed only IL-17RC. However, PBMCs expressed both IL-17RA and IL-17RC.</p

Effect of IL-17A, IL-17F or IL-22 on TER of cultured ARPE-19 monolayer.

<p>Monolayers were cultured for 21 days, where after the various stimuli were added. Incubation of ARPE-19 monolayers with 50 ng/ml IL-17A or IL-17F induced a gradual decrease of TER, and a significant effect occurred 5 days (p = 0.019, p = 0.045) after stimulation. The continuous decreases were also observed 6 days (p = 0.01, p = 0.016) and 7 days (p = 0.023, p = 0.008) after stimulation. IL-22 had no effect on TER. Data were shown as the means±SEM of four independent experiments. *<i>p</i><0.05 versus the control group.</p

Data_Sheet_1_Optical Coherence Tomographic Features and Prognostic Values of Macular Edema in Vogt-Koyanagi-Harada Disease.docx

Purpose: To determine optical coherence tomographic (OCT) features of macular edema (ME) and identify potential prognostic values for ME and visual outcomes in Vogt-Koyanagi-Harada disease (VKH).Methods: In the retrospective case series, a total of 1,377 VKH patients who were seen in a tertiary uveitis center between September 2011 and January 2018 were reviewed on their demographics, visual acuity, ocular and extraocular manifestations, modes of treatment, and OCT examinations. Of these patients, 79 (5.7%) having ME were included for analysis of OCT features. Four patients were missed without ME resolution, and the remaining 75 patients who either had ME resolved or were followed up for 2 years were included for analysis of disease outcomes.Results: Of the 115 affected eyes in these 79 patients, 100 (87.0%) had cystoid ME (CME), accounting for the most common OCT feature of VKH-related ME. Disruption of the inner-segment/outer-segment junction (IS/OS) band seen in 33 (28.7%) affected eyes of 24 (30.4%) patients was found as a risk factor for the development of persistent ME [10 of 62 (16.1%) vs. 13 of 13 (100%); P Conclusions: Intraretinal cystoid changes are most commonly seen in the edematous macula of VKH patients. Disruption of the IS/OS band is a useful risk sign for poor ME and visual outcomes in VKH-related ME, and a long-term well-controlled intraocular inflammation may be critical for the resolution of refractory cases.</p

The expression of IL-9 in the EAU mice and the control mice.

<p>Splenocytes and DLN cells, obtained from the EAU mice (inflammatory phase and recovery phase) or control mice (n = 5 per group), were activated with anti-CD3/D28 (1 µg/ml) (A) or IRBP_161–180 (20 µg/ml) (B) for 3 days. IL-9 was analyzed by ELISA. Splenocytes (C) and DLN cells (D), obtained from the immunized mice on indicated time points, were stimulated with IRBP_161–180 (20 µg/ml) for 3 days, and the supernatants were collected for measuring IL-9, IL-17 and IFN-γ. Data are representative of three independent experiments. ND: not detected.</p

Scheme of the AAV2 vector construct.

<p>The transgene is under the control of a CMV promoter and followed by BGH poly (A). The expression cassette is flanked by ITRs. CMV promoter, human cytomegalovirus immediate early promoter; hIFN-α, human interferon-alpha; BGH poly (A), BGH poly-adenylation signal; ITR, AAV2 inverted terminal repeats.</p

Effect of IFN-β treatment on IL-9 production.

<p>Splenocytes (A) and DLN cells (B) from the immunized mice following IFN-β or PBS treatment were activated with IRBP_161–180 (20 µg/ml) or anti-CD3/CD28 (1 µg/ml) for 3 days. IL-9 was analyzed by ELISA. Data are representative of three independent experiments with at least five mice per group. ND: not detected.</p

Effect of IFN-β on IL-9 production by polarized Th1, Th17 cells and effctor/memory T cells.

<p>Naïve T cells cultured with or without IFN-β in Th1-polarizing conditions (10 ng/ml IL-12) (A) or Th17-polarizing conditions (20 ng/ml IL-6, 5 ng/ml TGF-β1 and 10 ng/ml IL-23) (B) for 4 days. Effector/memory T cells were cultured with or without IFN-β for 3 days. IL-9 was analyzed by ELISA. Data are representative of three independent experiments.</p