Hongyan Zhou

Longitudinal quantification of aqueous flare and cells in Vogt–Koyanagi–Harada disease

Aims: To quantitatively evaluate the changes of aqueous flare and cells in eyes with Vogt–Koyanagi–Harada (VKH) disease. Methods: This prospective study included 35 initial-onset VKH patients (70 eyes) and 46 recurrent VKH patients (92 eyes) following immunotherapy. Aqueous flare and cells were quantified using the laser flare-cell meter before treatment, 2 weeks, 1, 3, 6 and 9 months after treatment. Results: Before treatment, mean aqueous flare (ph/ms) in initial-onset and recurrent VKH eyes were 8.1 (SD 4.1) vs 43.6 (20.7) (pu200a=u200a0.000). Following treatment, recurrent VKH eyes showed a significantly higher flare value than initial-onset VKH eyes at 2 weeks, 1, 3 and 6 months. Prior to treatment, mean cell counts (cells/0.5 mm3) in initial-onset and recurrent VKH eyes were 2.0 (1.9) vs 39.4 (23.1) (pu200a=u200a0.000). Following treatment, recurrent VKH eyes showed significantly higher cell counts than initial-onset VKH eyes at 2 weeks, 1 and 3 months. Conclusions: Our study shows that recurrent VKH patients displayed a more striking and long-lasting breakdown of the BAB and more severe inflammation than initial-onset VKH patients. Our study also indicates that the disruption of BAB lasted longer than aqueous cells either in initial-onset or in recurrent VKH patients.

Inhibitory effect of rapamycin and dexamethasone on production of IL-17 and IFN-gamma in Vogt-Koyanagi-Harada patients

Aims: To evaluate the effect of rapamycin (RAPA) and dexamethasone (DEX) on the production of IL-17 and IFN-γ by peripheral blood mononuclear cells (PBMCs) from Vogt–Koyanagi–Harada (VKH) patients and healthy individuals. Methods: Blood samples were drawn from 10 active VKH patients and 10 healthy individuals. PBMCs were cultured with or without anti-CD3 and anti-CD28 antibodies in the presence or absence of different concentrations of RAPA or DEX for 72 h. IL-17 and IFN-γ concentrations in the supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Results: The expression of IL-17 and IFN-γ was significantly increased in active VKH patients compared with that in healthy controls. Both RAPA and DEX were able to significantly inhibit the production of IL-17 and IFN-γ by PBMCs from patients and healthy controls. RAPA was able to completely inhibit IL-17 production at a dosage of 10 ng/ml but only partially suppressed IFN-γ production even at a much higher concentration (1000 ng/ml). DEX inhibited the production of both IL-17 and IFN-γ by approximately 70%. Conclusions: This study indicates that both RAPA and DEX inhibit the production of IL-17 and IFN-γ by PBMCs. RAPA is much stronger in inhibiting the production of IL-17 than DEX.

Splenic CD8+ T cells secrete TGF-β1 to exert suppression in mice with anterior chamber-associated immune deviation

BackgroundCD8+ regulatory T cells (Treg) have been considered to be involved in a model of ocular-induced tolerance, known as anterior chamber-associated immune deviation (ACAID). The mechanisms of suppression by CD8+ T cells in ACAID remain only poorly understood. TGF-β1 is considered as an inhibitory cytokine for immunosuppression in some models. The production of TGF-β1 by CD8+ T cells in ACAID, and whether CD8+ T cells exert suppression through TGF-β1, is unknown.MethodsThe suppressive effect of CD8+ T cells in ACAID mice was determined by a local adoptive transfer (LAT) assay. The production of TGF-β1 by CD8+ T cells was measured by enzyme-linked immunosorbent assay (ELISA). Anti-TGF-β1 antibodies were used in the LAT assay to test if they could block the inhibitory effect of CD8+ T cells.ResultsCD8+ T cells from ACAID mice were shown to block the delayed-type hypersensitivity (DTH) response in an antigen-specific manner in a LAT assay. These CD8+ T cells secreted TGF-β1, and their suppression could partially be blocked by anti-TGF-β1 antibodies.ConclusionsOur study confirms that CD8+ T cells from ACAID mice possess inhibitory properties. This population exerts part of its suppressive function via the production of TGF-β1.

MALDI-TOF/TOF-MS Reveals Elevated Serum Haptoglobin and Amyloid A in Behcet’s Disease

Behcets disease (BD) is a multisystemic autoimmune disease with unclear etiology and pathogenesis. To screen aberrant serum proteins in BD, serum samples were obtained from eight male BD patients with active uveitis and eight male healthy volunteers with informed consent. The serum samples from active BD patients and normal controls were pooled. Highly abundant serum proteins (albumin and IgG) were depleted from these two samples using an affinity capture based kit. The obtained samples were subjected to two-dimensional gel electrophoresis (2-DE). Protein spots were visualized with the blue silver staining. Differently expressed proteins were subsequently identified by matrix-assisted laser desorption /ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). Western blot and enzyme-linked immunosorbent assay (ELISA) were performed using the serum samples from 18 patients with active BD, 6 patients with inactive BD, 22 patients with Vogt-Koyanagi-Harada (VKH) syndrome, and 20 healthy volunteers to validate the results of 2-DE and MS. Proteomic profiles of the pooled samples were compared, and approximately 800 protein spots were observed in each of the gels. Expression levels of four of the protein spots in active BD were significantly higher than those in the normal controls. Mass spectrometric protein identification revealed that the four protein spots corresponded to two proteins: haptoglobin (Hp) and serum amyloid A (SAA). Western blot and ELISA showed that Hp was only overexpressed in active BD but not in inactive BD, VKH syndrome, or healthy controls. An obvious band of SAA was detected in 72.2% of the serum samples from BD patients, whereas a vague band of this protein was found in 10.0% of the tested normal samples and 9.1% of VKH samples. Our results revealed a significantly increased expression of Hp and SAA in serum of active BD patients. These two proteins may be involved in the development of BD.

Disturbed expression of Fas/FasL on CD4(+) and CD8(+)T cells in Behcet's disease, Vogt-Koyanagi-Harada syndrome, and idiopathic anterior uveitis.

Aims: To evaluate the expression of Fas/FasL antigen on peripheral blood T lymphocytes in patients with Behcets disease, Vogt-Koyanagi-Harada (VKH) syndrome, and idiopathic anterior uveitis. Methods: The expression of Fas and FasL on peripheral blood T lymphocytes was determined using flow cytometry in 26 patients with Behcets disease (BD), 17 patients with VKH syndrome, 25 patients with idiopathic anterior uveitis, and 43 healthy individuals (controls). Results: A higher proportion of CD4 + T cells expressing Fas was noted in patients with Behcets disease (25.70 ± 7.32%), VKH syndrome (19.60 ± 11.02%), and idiopathic anterior uveitis (20.81 ± 7.40%) compared with controls (14.02 ± 6.30%). The expression of Fas on CD8 + cells from patients with Behcets disease (9.47 ± 6.97%) and VKH syndrome (6.84 ± 5.5%) was also higher than that seen in controls (3.47 ± 2.75%). There was no difference in FasL expression on T cells between patients and controls except that a lower expression of FasL on CD8+ T cells was noted in patients with idiopathic anterior uveitis. Conclusion: A disturbed expression of Fas and FasL on T cells is present in patients with Behcets disease, VKH syndrome, and idiopathic anterior uveitis, which may be involved in the perpetuation and recurrence of uveitis.

Upregulation of CD94 on CD8+T Cells in Anterior Chamber-Associated Immune Deviation

BackgroundCD8+ regulatory T cells (Treg) have been considered to be involved in a model of ocular-induced tolerance, known as anterior chamber-associated immune deviation (ACAID). The phenotype and characteristics of CD8+Treg in ACAID remain only poorly understood. Recent studies have reported that the CD94-Qa-1 system is implicated in the induction of ACAID CD8+Treg, but the functions and characteristics of CD8+CD94+T cells remain unclear.ResultsBoth mRNA and protein of CD94 and NKG2A were markedly up-regulated on splenic CD8+T cells of ACAID mice compared with controls. Flow cytometric analysis showed that very few CD8+CD94+T cells express granzyme B, perforin and Foxp3. CD8+CD94+T cells, but not CD8+CD94-T cells, magnetically isolated from the spleens of ACAID mice, produced large amounts of TGF-beta1 and exhibited suppressive activity in vitro. Neutralization of TGF-beta1 caused reversal of suppression mediated by CD8+CD94+T cells.ConclusionCD8+CD94+T cells from ACAID mice exhibited suppressive activity in association with enhanced expression of TGF-beta1, suggesting that CD8+Treg are mainly distributed in CD94+T cell subpopulations.

No association of CTLA-4 polymorphisms with susceptibility to Behçet disease.

Background: Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) is a key negative regulator of T lymphocytes and has been shown to be associated with a number of autoimmune diseases. The present study was performed to assess the association between CTLA-4 polymorphisms and Behçet disease (BD) in Chinese patients. Methods: Two hundred and twenty-eight BD patients and 207 controls were analysed for four single nucleotide polymorphisms (SNPs) (−1661A/G, −318C/T, +49G/A and CT60G/A) in the CTLA-4 gene by PCR-restriction fragment length polymorphism (RFLP) analysis. The association between SNP +49A/G and BD in Chinese population as well as other ethnic groups was analysed by meta-analysis. Results: No association could be detected between CTLA-4 SNPs or haplotypes and BD. Also, no association was observed between CTLA-4 polymorphisms and BD subgroups, stratified by clinical features. A meta-analysis showed that there was no heterogeneity between studies (pu200a=u200a0.60, I2u200a=u200a0%) and that CTLA-4 SNP +49 was not associated with BD (overall effect: Zu200a=u200a0.26, pu200a=u200a0.79). Conclusion: This study and a meta-analysis failed to demonstrate any association between the tested CTLA-4 polymorphisms and BD.