Fulya Cosan

Genome-wide association study identifies variants in the MHC class I, IL10, and IL23R-IL12RB2 regions associated with Behcet's disease

Behçets disease is a genetically complex disease of unknown etiology characterized by recurrent inflammatory attacks affecting the orogenital mucosa, eyes and skin. We performed a genome-wide association study with 311,459 SNPs in 1,215 individuals with Behçets disease (cases) and 1,278 healthy controls from Turkey. We confirmed the known association of Behçets disease with HLA-B*51 and identified a second, independent association within the MHC Class I region. We also identified an association at IL10 (rs1518111, P = 1.88 × 10−8). Using a meta-analysis with an additional five cohorts from Turkey, the Middle East, Europe and Asia, comprising a total of 2,430 cases and 2,660 controls, we identified associations at IL10 (rs1518111, P = 3.54 × 10−18, odds ratio = 1.45, 95% CI 1.34–1.58) and the IL23R-IL12RB2 locus (rs924080, P = 6.69 × 10−9, OR = 1.28, 95% CI 1.18–1.39). The disease-associated IL10 variant (the rs1518111 A allele) was associated with diminished mRNA expression and low protein production.

Association of Familial Mediterranean Fever-Related MEFV Variations With Ankylosing Spondylitis

OBJECTIVE The pathogenesis of ankylosing spondylitis (AS) has a strong genetic contribution. Familial Mediterranean fever (FMF) is an autosomal recessively inherited autoinflammatory disorder caused by MEFV gene missense variations, and a clinical association between FMF and AS has been reported previously. The aim of this study was to analyze the association of common MEFV variations (M694V, M680I, V726A, and E148Q) with AS in a group of Turkish patients. METHODS The study group comprised 193 patients with AS and 103 matched healthy control subjects. All individuals were genotyped for 4 MEFV variations and HLA-B27 using genomic DNA, and association of the variations with the clinical and laboratory features of the patients was analyzed. RESULTS The MEFV missense variations were significantly more frequent in patients with AS (22.3%) compared with healthy control subjects (9.7%; odds ratio [OR] 2.67, 95% confidence interval [95% CI] 1.28-5.56). This difference was more prominent for exon 10 variations (M694V, V726A, M680I) (OR 3.75, 95% CI 1.41-9.97), especially for the most-penetrant variation M694V (OR 4.73, 95% CI 1.39-16.12). MEFV variations were more frequent in HLA-B27-negative patients with AS, and the difference was statistically significant in patients carrying exon 10 variants. CONCLUSION FMF-related MEFV variations are associated with AS, and these variations may contribute to the pathogenesis of AS, especially in populations in which the prevalence of FMF is high.

Prevalence and virological features of occult hepatitis B virus infection in female sex workers who work uncontrolled in Turkey.

Background: There is little information about the prevalence of occult hepatitis B virus infection (OHBVI). We have investigated the prevalence and virological features of OHBVI among female sex workers (FSWs) in Istanbul.

IL-22-secreting Th22 and IFN-γ-secreting Th17 cells in Behçet's disease

Abstract Objective. Behçets disease (BD) is a systemic inflammatory disease with unknown etiology. Studies have shown that some T helper (Th) 1-associated cytokines have role in the inflammation of BD. The CD4+ Th cells can be differentiated into Th1, Th2, Th17 and Th22 secrete different cytokines to regulate immune system. In this study, cytokine secretion of Th subsets in BD was investigated. Methods. The study group consisted of 26 BD patients with mucocutaneous involvement and 12 healthy subjects. Lymphocyte subpopulations, IL-5, IL-10, IL-17, IL-22 and IFN-γ secretion of CD4+ T and Foxp3+ Treg cells were determined by flow cytometry. Results. Compared with healthy subjects, Th1 (IL-17A−IL-22−IFN-γ+), Th22 (IL-17A−IL-22+IFN-γ) and IL-17A+IFN-γ+-secreting cells were significantly increased, and the percentage of Treg cells were dramatically reduced in BD patients. The frequency of recurrent oral ulcers was associated with increased Th22 cells. Conclusions. Our study describes an association between Th22 cell subset and IL-17A+ IFNγ+-secreting cells with mucocutaneous BD. These findings revealed that reduced levels of Tregs and increased levels of Th1 and Th22 cells as well as Th17/Th1 cells might be associated with the pathogenesis of BD.

Behçet’s disease: immunological relevance with arthritis of ankylosing spondylitis

Behçet’s disease (BD) is a multi-system inflammatory disorder, in which cytokine balance is polarized to Th1. In this study, the cell surface molecule expression, Th1/Th2, inflammatory cytokine levels in blood, and synovial fluid of CD3+ T lymphocytes in BD were investigated. The study group consisted of 10 BD, 10 ankylosing spondylitis (AS) patients with peripheral arthritis, and 10 healthy subjects. Expression of cell surface molecules, intracellular IL-2, IL-5, IL-8, IL-10, IL-12, IFN-γ, and TNF-α levels in CD3+ T lymphocytes were determined by flow cytometry in synovial and peripheral blood mononuclear cells (PBMCs). Synovial and plasma cytokine levels were measured by ELISA and CBA. In PBMCs, CD4, CD25, HLA-DR expression and intracellular IL-12, and TNF-α levels of CD3+ T lymphocytes were statistically increased in BD patients compared to healthy subjects. Compare to AS patients, CD25 and HLA-DR surface expression and intracellular IFN-γ and TNF-α levels in T cells were significantly elevated in BD patients. In BD patients, there was an increase in IL-8 secretion; however, in AS patients, both Th1- and Th2-type cytokines were increased compare to healthy subjects. Intracellular cytokine expression did not show any difference in BD patients; however, IL-12 content of synovial fluid was significantly increased compared to AS patients. Our findings revealed that Th1 polarization occurred in both peripheral blood and synovial fluid of BD patients with arthritis. It is found no difference between synovial fluid analysis of BD and AS patients, showing the similarities in the pathogenesis of both diseases.

A genome-wide analysis of lentivector integration sites using targeted sequence capture and next generation sequencing technology

One application of next-generation sequencing (NGS) is the targeted resequencing of interested genes which has not been used in viral integration site analysis of gene therapy applications. Here, we combined targeted sequence capture array and next generation sequencing to address the whole genome profiling of viral integration sites. Human 293T and K562 cells were transduced with a HIV-1 derived vector. A custom made DNA probe sets targeted pLVTHM vector used to capture lentiviral vector/human genome junctions. The captured DNA was sequenced using GS FLX platform. Seven thousand four hundred and eighty four human genome sequences flanking the long terminal repeats (LTR) of pLVTHM fragment sequences matched with an identity of at least 98% and minimum 50 bp criteria in both cells. In total, 203 unique integration sites were identified. The integrations in both cell lines were totally distant from the CpG islands and from the transcription start sites and preferentially located in introns. A comparison between the two cell lines showed that the lentiviral-transduced DNA does not have the same preferred regions in the two different cell lines.

No association of the TLR2 gene Arg753Gln polymorphism with rheumatic heart disease and Behçet’s disease

Behçet’s disease (BD) is a multisystem inflammatory disorder of unknown etiology, and infections with different microorganisms including streptococci have been claimed as triggers of inflammatory attacks in BD pathogenesis. Toll-like receptor 2 (TLR2) has been known to recognize several microbial antigens including that of streptococci, and TLR2 gene Arg753Gln polymorphism has been reported to be strongly associated with acute rheumatic fever with an odds ratio of 100. This study aimed to investigate the TLR2 gene Arg753Gln polymorphism in a group of patients with BD and rheumatic heart disease (RHD) and to analyze the role of genotyping errors resulting from duplicated gene segments. The study group consisted of 211 patients with BD, 95 patients with RHD, and 94 matched Turkish healthy controls. Because of the duplicated exon 3 in 23-kb upstream of the TLR2 gene, genotyping for the Arg753Gln polymorphism with polymerase chain reaction–restriction fragment length polymorphism method was carried out using a new set of primers and PstI restriction enzyme. TLR2 gene Gln753 allele was observed in two of 211 (1.0%) patients with BD, five of 95 (5.3%) patients with RHD, and two of 94 (2.1%) healthy controls. All patients and controls were found to be heterozygous for Arg753Gln polymorphism, except one patient with BD, who was homozygous for Gln753. Although a slight increase of heterozygosity was noted in patients with RHD, no statistically significant difference was observed in the distribution of Arg753Gln polymorphism in BD and RHD compared to healthy controls. In conclusion, TLR2 gene Arg753Gln polymorphism is not associated with BD nor with RHD; and a duplicated region of the TLR2 exon 3 located 23-kb upstream of the polymorphic region may explain contradictory association findings described so far.

Sinus node dysfunction in adult systemic lupus erythematosus flare: A case report.

Abstract Cardiac involvement can affect up to 50% of the systemic lupus erythematosus (SLE) patients but conduction system disturbances in SLE are less commonly described. For an early detection of this complication in the acute phase of SLE a whole cardiovascular examination and periodic electrocardiographic monitoring are recommended. We describe a patient who was diagnosed with flare up of lupus activity manifesting as sinus node dysfunction presenting as profound sinus bradycardia. She was successfully treated with high-dose methylprednisolone therapy.

Bilateral acute sacroiliitis due to isotretinoin therapy: a case report

Dear Editor, We report here a case of a young female patient who developed sacroiliitis related to isotretinoin (13-cisretinoic acid) use which completely resolved following discontinuation of the medication. Isotretinoin, a retinoid indicated for the treatment of severe cystic acne, has been associated with adverse effects, including musculoskeletal symptoms such as arthritis, arthralgia, myalgia and soft tissue calcification. Reactive sacroiliitis is a rare side effect. A small number of cases of sacroiliitis occurring in association with isotretinoin use have been described. A 20-year-old healthy woman was admitted to our department with complaints of myalgia, bilateral hip, pelvic and low back pain. She was not using any drugs except isotretinoin, which she had been using to treat acne for the previous 3 months (initial dose 30 mg/day for the first month, then increased to 40 mg/day for the following 2 months). Because of increased pain, the patient had stopped using the drug 15 days ago. On physical examination, sacroiliac joints were very painful and Gaenslen, Thigh Thrust, Patrick Faber and Mennel tests were strongly positive. Muscle power was normal (Medical Research Council scale 5/5). Laboratory examination showed that the erythrocyte sedimentation rate was 52 mm/h (normal range 0–20), C-reactive protein 2.75 mg/dL (0.0–0.5), creatinekinase (CK) 43 U/L (30–200). Antinuclear antibodies, antineutrophil cytoplasmic antibodies, rheumatoid factor, C3, C4 were all negative. Blood chemistry, thyroid function tests and complete blood count were unremarkable. Screening for viruses and brucella was negative. Human leukocyte antigen (HLA)-B27 was positive. X-ray of pelvis and sacroiliac joint was normal. Magnetic resonance imaging (MRI) showed bilateral active inflammatory sacroiliitis with evidence of bone marrow edema adjacent to both sacroiliac joints. In dynamic contrast enhanced MRI, Fehn activity was measured 103% around the right sacroiliac joint and 75% around the left sacroiliac joint. These measures were compatible with prominent activity (Fig. 1). The patient was treated with prednisolone (15 mg daily slowly decreasing until suspension) and diclofenac sodium (75 mg daily) with gradual improvement over 2 months. After 6 weeks, the symptoms had completely resolved and the laboratory results had improved, so the treatment was stopped. There was no complaint and the laboratory results were normal on the sixth month of follow-up. The use of the Naranjo probability scale indicated a probable relationship between isotretinoin therapy and sacroiliitis. Isotretinoin is a retinoid that is used to treat cystic acne, comedonal acne and other diseases. Its most common side effects are mucocutaneous and ocular in nature (i.e., cheilitis, ocular sicca and decreased dark adaptation). Reactive sacroiliitis is a rare side effect. On physical examination and MRI findings, we see the symptoms related to sacroiliitis which are men-

The association between P selectin glycoprotein ligand 1 gene variable number of tandem repeats polymorphism and risk of thrombosis in Behçet's disease

Behçets disease (BD) has been recognized as an unclassified type of vasculitis with an accompanying tendency to thrombosis. No disease‐specific pathology has been demonstrated so far to explain the prothrombotic state, and this predisposition is considered to be associated with endothelial activation/dysfunction. P‐selectin glycoprotein ligand‐1 (PSGL‐1) variable number of tandem repeat (VNTR) polymorphism has an impact on the protein length, and heterozygosity affect of the PSGL‐1 to P‐selectin interaction, which has been found to be associated with an increased risk of thrombosis in patients with antiphospholipid syndrome. We aimed to analyze the association of PSGL‐1 gene polymorphism, in a group of BD patients with and without thrombosis.