Yukihiro Horie

Genome-wide association studies identify IL23R - IL12RB2 and IL10 as Behçet's disease susceptibility loci

Behçets disease is a chronic systemic inflammatory disorder characterized by four major manifestations: recurrent ocular symptoms, oral and genital ulcers and skin lesions. We conducted a genome-wide association study in a Japanese cohort including 612 individuals with Behçets disease and 740 unaffected individuals (controls). We identified two suggestive associations on chromosomes 1p31.3 (IL23R-IL12RB2, rs12119179, P = 2.7 × 10−8) and 1q32.1 (IL10, rs1554286, P = 8.0 × 10−8). A meta-analysis of these two loci with results from additional Turkish and Korean cohorts showed genome-wide significant associations (rs1495965 in IL23R-IL12RB2, P = 1.9 × 10−11, odds ratio = 1.35; rs1800871 in IL10, P = 1.0 × 10−14, odds ratio = 1.45).

Genome-wide association study identifies GIMAP as a novel susceptibility locus for Behçet's disease

Objectives To identify non-major histocompatibility complex susceptible genes that might contribute to Behçets disease (BD). Methods We performed a genome-wide association study using DNA samples from a Korean population consisting of 379 BD patients and 800 controls. A replication study was performed in a Japanese population (363 BD patients and 272 controls). To evaluate the functional implication of the target single nucleotide polymorphisms (SNP), gene expression levels in peripheral T cells, allele-specific modulation of promoter activity and biological effect of mRNA knockdown were investigated. Results We found a novel association of BD to the GIMAP locus, mapped to chromosome 7q36.1 (rs1608157, p=6.01×10−8 in a minor allele dominant model; rs11769828, allele based p=1.60×10−6). A fine mapping study identified an association with four additional SNP: rs1522596 (OR=1.45, p=7.70×10−6) in GIMAP4; rs10266069 (OR=1.32, p=2.67×10−4) and rs10256482 (OR=1.27, p=5.27×10−4) in GIMAP2; and rs2286900 (OR=1.61, p=3.53×10−5) in GIMAP1 areas. Replication study using DNA samples from the Japanese population validated the significant association between BD and the GIMAP locus. The GIMAP4 promoter construct plasmid with the minor allele of rs1608157 displayed significantly lower activity than one with the major allele. Moreover, CD4 T cells from BD patients showed a lower level of GIMAP4 mRNA, and GIMAP4 knockdown was protective against Fas-mediated apoptosis. Conclusions These results suggest that a GIMAP cluster is a novel susceptibility locus for BD, which is involved in T-cell survival, and T-cell aberration can contribute to the development of BD.

Association of TLR4 polymorphisms with Behçet's disease in a Korean population

OBJECTIVES HLA-B51 is strongly associated with Behçets disease (BD) in any ethnic background. We recently reported that another gene, Toll-like receptor-4 (TLR4) is also implicated in BD in a Japanese population. To confirm these results, we investigated polymorphisms in the TLR4 gene in Korean patients with BD. METHODS In this study, 119 patients with BD and 141 healthy controls were enrolled; every participant was a Korean. Nine single nucleotide polymorphisms previously detected in TLR4 by direct sequencing were analysed for an association with BD. RESULTS The most frequent haplotype, TAGCGGTAA, was significantly increased in HLA-B*51-positive BD patients (49.5%), compared with healthy control participants [32.3%; P = 0.029; odds ratio (OR) = 2.01; 95% CI 1.25-3.23]. This haplotype was also significantly increased in BD patients with arthritis (48.2%; P = 0.003; OR = 1.96; 95% CI 1.26-3.26). There were no significant differences in the allele and genotype frequencies of patients and controls for each single nucleotide polymorphism. CONCLUSIONS The haplotype of TLR4 may increase the risk for developing BD and the complication of arthritis in the Korean population.

Investigation of Association between TLR9 Gene Polymorphisms and VKH in Japanese Patients

Purpose: Vogt-Koyanagi-Harada (VKH) disease is an autoimmune disorder affecting melanocytes in the skin, eyes, inner ear, and meninges. The Epstein-Barr virus and cytomegalovirus (CMV) antigen have been hypothesized as possible triggering factors for the disease. Toll-like receptors (TLRs) play an important role in the induction of defense mechanisms of the innate and adaptive immune responses to microbial pathogens. Among TLRs, TLR9 recognizes unmethylated 2′-deoxyribo (cytidine-phosphate guanosine)(CpG) DNA motifs that are frequently present in viruses and plays a central role in the host defense against viral infection. The aim of this study was to investigate whether TLR9 polymorphisms were associated with VKH in a Japanese population. Methods: Ninety-four Japanese patients diagnosed with VKH and 125 healthy control subjects were recruited. Five single-nucleotide polymorphisms (SNPs: rs187084, rs5743836, rs352139, rs352140, rs5743845) in the TLR9 gene were genotyped, and allelic and phenotypic diversity was assessed between cases and control subjects. Results: Strong linkage disequilibrium was observed among three SNPs (D’ > 0.99), which were located in one haplotype block. Two SNPs (rs5743836 and rs5743845) were monopolymorphic in both cases and controls. No statistically significant association was observed for any of the SNPs between cases and controls. Conclusion: Three SNPs in the TLR9 gene were not significantly associated with susceptibility to VKH.

Induction of Heat Shock Protein 70 Ameliorates Ultraviolet-Induced Photokeratitis in Mice

Acute ultraviolet (UV) B exposure causes photokeratitis and induces apoptosis in corneal cells. Geranylgeranylacetone (GGA) is an acyclic polyisoprenoid that induces expression of heat shock protein (HSP)70, a soluble intracellular chaperone protein expressed in various tissues, protecting cells against stress conditions. We examined whether induction of HSP70 has therapeutic effects on UV-photokeratitis in mice. C57 BL/6 mice were divided into four groups, GGA-treated (500 mg/kg/mouse) and UVB-exposed (400 mJ/cm2), GGA-untreated UVB-exposed (400 mJ/cm2), GGA-treated (500 mg/kg/mouse) but not exposed and naive controls. Eyeballs were collected 24 h after irradiation, and corneas were stained with hematoxylin and eosin (H&E) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). HSP70, reactive oxygen species (ROS) production, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and protein kinase B (Akt) expression were also evaluated. Irradiated corneal epithelium was significantly thicker in the eyes of mice treated with GGA compared with those given the vehicle alone (p < 0.01). Significantly fewer TUNEL-positive cells were observed in the eyes of GGA-treated mice than controls after irradiation (p < 0.01). Corneal HSP70 levels were significantly elevated in corneas of mice treated with GGA (p < 0.05). ROS signal was not affected by GGA. NF-κB activation was reduced but phospho-(Ser/Ther) Akt substrate expression was increased in corneas after irradiation when treated with GGA. GGA-treatment induced HSP70 expression and ameliorated UV-induced corneal damage through the reduced NF-κB activation and possibly increased Akt phosphorilation.

HLA-B51 Carriers are Susceptible to Ocular Symptoms of Behçet Disease and the Association between the Two Becomes Stronger towards the East along the Silk Road: A Literature Survey

ABSTRACT Purpose: Behçet disease (BD) is predominantly found between East Asia and the Mediterranean basin along the historic Silk Road. HLA-B51 is known to be strongly associated with BD. We investigated the association between HLA-B51 and the ocular manifestations of BD among various ethnic groups. Methods: A literature survey was conducted, and 18 articles written in English were reviewed. Results: A strong correlation was found between HLA-B51 and ocular lesions in the entire cohort discussed in the reviewed articles (OR = 1.76, p = 0.000057). HLA-B51 was shown to have a strong association with ocular manifestations of BD patients in East-Eurasian (OR = 2.40, p = 0.0030) and Middle-Eurasian (OR = 1.87, p = 0.0045), but not in West-Eurasian (OR = 1.28, p = 0.35) areas. This correlation seemed to become stronger towards the east. Conclusions: A meta-analysis showed that the correlation became stronger towards the east along the Silk Road. The study results may facilitate understanding of the etiology and characteristics of BD.

Replication of a microsatellite genome-wide association study of Behçet's disease in a Korean population

OBJECTIVE Behçets disease is one of the major aetiologies of uveitis causing blindness in Asian countries. A genome-wide association study identified six microsatellite markers as disease susceptibility loci for Japanese patients with Behçets disease. To confirm our recent results, these microsatellite markers were examined in a Korean population as a replication study. METHODS Study participants included 119 Behçets disease patients and 141 controls. All were enrolled in Korea. Association between the six reported microsatellite markers (D3S0186i, D6S0014i, D6S0032i, 536G12A, D12S0645i and D22S0104i) and Behçets disease was analysed. HLA-B was genotyped by sequence-based typing methods. RESULTS A microsatellite marker located near the HLA-B region demonstrated significant association with Behçets disease (P = 0.028). The genotype and phenotype frequencies of the HLA-B*51 gene were significantly increased in patients (23.1 and 39.5%, respectively) compared with healthy controls (11.2 and 20.1%, respectively; P < 0.001). CONCLUSION Microsatellite analysis revealed that the HLA-B*51 gene was strongly associated with Behçets disease in a Korean population.

Prompt therapy reduces the duration of systemic corticosteroids in Vogt-Koyanagi-Harada disease

Dear Editor, Vogt-Koyanagi-Harada (VKH) disease is the second most common disease diagnosis among Japanese people [1]. The disease is more commonly seen in Far East Asians, Hispanics, and Native Americans than in Caucasian and African people. The disease is believed to be a cellmediated autoimmune disorder affecting melanocytes, thus, any organs with melanocytes present are affected in the disease, e.g., eyes, skin, and meninges. The patients often suffer from granulomatous uveitis, poliosis, vitiligo, alopecia, auditory signs, and central nervous system symptoms. Profiles of cell surface markers were quite similar for cerebrospinal fluid and aqueous humor; the number of CD3, CD4, and CD4CD45RO cells was significantly greater in aqueous humor and cerebrospinal fluid than in peripheral blood lymphocytes [3]. Systemic corticosteroids are commonly used for the treatment. Though the majority of VKH patients have a favorable visual prognosis, chronic recurrent intraocular inflammation occurs in some cases, despite the use of high-dose corticosteroids systemically. To examine the prognostic factors in VKH disease, correlation between the timing of the initiation of systemic corticosteroid and the total period of corticosteroid administration was retrospectively compared. In the present study, all patients were Japanese. They were diagnosed as VKH disease at the Uveitis Survey Clinic of the Hokkaido University Hospital. Diagnosis of VKH disease was based on the revised criteria laid down by the International Committee in 2001 and traditional Sugiura’s criteria [4]. These two criteria corresponded well in patients of the present study. We retrospectively examined the medical records of 65 uveitis patients with VKH focusing on the duration of systemic corticosteroid therapy. All patients were treated with prednisolone at 200 mg/day as an initial dose at our department in the hospital. Dosage of prednisolone was gradually tapered in each patient according to the clinical effects. Patients were divided into two groups according to the timing of start of the therapy. Patients receiving systemic corticosteroid therapy within 13 days after the disease onset were classified as the “Early” group, and those receiving on day 14 or later were classified as the “Late” group. It was shown that the “Early” group received systemic corticosteroids for 10.9 (1.71, standard error, SE) months, whereas “Late” group did for 24.2 (4.58) months on average (Fig. 1). The mean duration of drug administration was significantly shorter in patients with early therapy group than those of late therapy group (p=0.0098). Visual outcome was good in both groups. Only three cases had visual acuity less than 0.1 in one side; poor visual acuity of a patient was caused by glaucoma from the Early group, and that of two cases of the Late group were attributed to cataract. Three decades ago, Ohno et al. divided VKH patients into two groups whose initiation of systemic corticosteroids were ≤30 and ≥31 days after disease onset among American people [5]. It was shown that patients who received systemic corticosteroids early in the course of the disease had a lesser incidence of multi-systemic involvement than those who received at a later date [5]. Since most of the Graefes Arch Clin Exp Ophthalmol (2008) 246:1641–1642 DOI 10.1007/s00417-008-0869-5

Evaluation of NLRP1 gene polymorphisms in Vogt-Koyanagi-Harada disease

PurposePolymorphisms of the NACHT [neuronal apoptosis inhibitory protein (NAIP), CIITA, HET-E, TP1] and leucine-rich repeat protein 1 (NLRP1) gene are reported to be associated with susceptibility to vitiligo and several autoimmune diseases. Vogt-Koyanagi-Harada (VKH) disease is an autoimmune disorder affecting melanocytes in the skin, eyes, inner ear, and meninges. In this study, genetic associations between VKH disease and single-nucleotide polymorphisms (SNPs) surrounding the NLRP1 gene were investigated.MethodsSix SNPs (rs6502867, rs925597, rs3926687, rs2733359, rs878329, and rs4790796) near the NLRP1 gene, including noncoding regions, were sequenced by a direct method to genotype 167 Japanese patients with VKH disease and 187 healthy Japanese volunteers.ResultsNone of the six SNPs in the NLRP1 region were significantly associated with disease susceptibility or the ocular, neurological, and dermatological manifestations of VKH.ConclusionsAlthough skin manifestations are clinically similar between vitiligo and VKH disease, the genetic and immunological mechanisms of these two diseases may be different.

Sister cases of Behcet’s disease and Vogt–Koyanagi–Harada disease

Behcet’s disease is an inflammatory systemic disease characterised by four major symptoms such as oral aphthous ulcers, genital ulcers, skin lesions, and intraocular inflammation.1 Vogt–Koyanagi–Harada (VKH) disease is also associated with panuveitis as an ocular sign, and is accompanied by meningismus, cerebrospinal fluid pleocytosis, auditory findings and integumentary findings.2 Genetic factors are implicated as one of the pathogenetic signs of both diseases.3 Some patients have risk alleles such as human leukocyte antigen (HLA)-B51 and DR4 but others do not. We experienced sister cases having the same HLA haplotypes; the elder sister became ill with Behcet’s disease and the younger sister was affected with VKH disease. A 54-year-old woman was referred to our hospital complaining of blurred vision in the left eye in 2002. She had had a history of oral aphthous ulcers for 40 years, genital ulcers and erythema nodosum for 15 years. Hypopyon iridocyclitis and retinal vasculitis with subretinal …