The inhibitory effect of IRAK1/4 inhibitor on the proliferation of CD4⁺T cells purified from PBMCs obtained from active VKH patients (n = 10) and normal controls (n = 11).

<p>Cells were cultured with IRAK1/4 inhibitor (50 nM) in the presence of IL-18 (100 ng/ml) or IL-1β (20 ng/ml) for 3 days. Anti-CD3 mAb (0.5 μg/ml) and anti-CD28 mAb (0.1 μg/ml) were added to the culture system at the same time. Proliferation of cultured CD4⁺T cells was tested by a modified MTT assay. Results are expressed as means ± SD.</p

The mRAN levels of IRAK1 and IRAK4 in active VKH patients, inactive VKH patients and normal controls.

<p><b>A</b>. The mRNA level in PBMCs of IRAK1 of active VKH patients (n = 18), inactive VKH patients (n = 12), and normal subjects (n = 20) was detected by real-time quantitative PCR. <b>B</b>. The mRNA level in PBMCs of IRAK4 of active VKH patients (n = 12), inactive VKH patients (n = 8), and normal subjects (n = 10) was detected by real-time quantitative PCR.</p

Inhibitory effect of IRAK1/4 inhibitor on the secretion of IFN-γ and IL-17 by CD4⁺T cells from active VKH patients (n = 10) and normal controls (n = 11).

<p>Cells were cultured with or without IRAK1/4 inhibitor in the presence of anti-CD3 and anti-CD28 antibodies for 3 days. Meanwhile, IL-18 or IL-1β was also added to the cultures. The level of IFN-γ or IL-17 was measured by ELISA. A. In the presence of IL-18, IFN-γ production by CD4⁺T cells from patients with active VKH disease (n = 10) and normal controls (n = 11). Results are expressed as means ± SD. B. In the presence of IL-1β, IL-17 production by CD4⁺T cells was measured in patients with active VKH disease (n = 10) and normal controls (n = 11). Results are expressed as means ± SD.</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

The effect of IRAK1/4 inhibitor on the expansion of Th1 cells and Th17 cells.

<p>Purified CD4⁺T cells from normal controls (n = 8) were cultured with or without IRAK1/4 inhibitor for 3 days in the presence of wither IL-18 or IL-1β. The frequency of Th1 and Th17 cells was analyzed by FCM. <b>A</b>. A representative patient with data near the mean of each group in B and C. <b>B</b>. The results represent the percentages of IFN-γ⁺, IL-17⁺, and IL-17⁺IFN-γ⁺ cells among the CD4⁺T cells in the presence of IL-18. Results are expressed as means ± SD. <b>C</b>. The results represent the percentages of IFN-γ⁺,IL-17⁺, and IL-17⁺IFN-γ⁺ cells among the CD4⁺T cells in the presence of IL-1β. Results are expressed as means ± SD.</p

The effect of IRAK1/4 inhibitor on the activities of STAT1 and STAT3.

<p>Purified CD4⁺T cells from normal controls (n = 6) were cultured with or without IRAK1/4 inhibitor in the presence of anti-CD3 and anti-CD28 antibodies for 30 minutes. The phosphorylation of STAT1 and STAT3 were analyzed by FCM. <b>A</b>. A representative patient with data near the mean of each group in groups B and C. <b>B</b>. The results represent the percentage of pSTAT1 and pSTAT3 among the CD4⁺T cells and (<b>C</b>) the intensity of pSTAT1 and pSTAT3 in the CD4⁺T cells. Results are expressed as means ± SD.</p

The effect of IRAK1/4 inhibitor on the phosphorylation of NF-κB.

<p>Purified CD4⁺T cells from normal controls (n = 6) were cultured with or without IRAK1/4 inhibitor in the presence of anti-CD3 and anti-CD28 antibodies for 30 minutes. <b>A</b>. A representative example with data near the mean of each group in group B. <b>B</b>. The results represent the mean fluorescence intensity of phosphorylation NF-κB expression in the CD4⁺T cells. Results are expressed as means ± SD.</p

Induction of EAU and analysis of the expression of IFN-β in this model.

<p>EAU was induced in B10RIII mice by immunization with 50 µg IRBP_161-180 in CFA. (A) Inflammatory infiltration in the anterior segment of the eye (after mydriasis). (B) Damaged retinal structure in a B10RIII mouse after immunization with IRBP_161-180. Eyes were obtained 13 days after immunization and stained with haematoxylin/eosin. (C) Mean histology score during the development of EAU. (D) Splenocytes from normal mice (day 0) and those obtained at different time points after the induction of EAU were stimulated by LPS for 24 hours. Supernatants were measured for the production of IFN-β by ELISA. Data are representative of three independent experiments. **p<0.01, *p<0.05, NS = not significant.</p

Effect of IFN-β on IL-6, TGF-β, IL-23 and IL-10 production by monocytes.

<p>Monocytes from immunized mice on day 13 (or on day 0 and day 16 where indicated), cultured with or without IFN-β (250 U/ml) for 24 hours, were stimulated with LPS (100 ng/ml) for another 24 hours. Cytokines were detected by ELISA. Data are representative of three independent experiments. **p<0.01, NS = not significant.</p

Association Between HLA Polymorphisms and Sympathetic Ophthalmia in Han Chinese

Sympathetic ophthalmia (SO) is considered as an autoimmune disease with unclear mechanisms. This study investigated the relationship between HLA polymorphisms and SO. HLA typing was performed using the LABType reverse SSO DNA typing method. The allele and haplotype frequencies were assessed using the PyPop software. Statistical significance of genotype distributions between 116 patients and 84 healthy individuals (control) was determined using Fisher’s exact test or Pearson's chi-squared test. The SO group had a higher frequency of HLA-DRB1 * 04:05, HLA-DQB1 * 04:01, DRB1 * 04:05-DQB1 * 04:01 haplotype   as compared to the control group (Pc  This study revealed that DRB1 * 04:05 and DQB1 * 04:01 alleles, as well as DRB1 * 04:05-DQB1 * 04:01 haplotye could be potential risk factors for SO.</p