Suzan Dinkci

Analysis of CD28 and CTLA‐4 gene polymorphisms in Turkish patients with Behcet's disease

In this study we aimed to investigate IVS3 +17T/C single nucleotide polymorphism (SNP) of CD28 gene, +49A/G and −318C/T SNPs of CTLA‐4 gene in patients with Behçets disease (BD) and their potential association to the main clinical features of the disease. These polymorphisms were investigated in a Turkish population of 123 patients with BD and 179 healthy controls, by using PCR‐RFLP technique. HLA‐B*51 genotype was also studied in both groups by using PCR‐SSP. The frequency of IVS3 +17TC genotype of the CD28 gene was significantly increased in BD patients compared to controls (43.6% vs. 31.2%, OR = 1.663, 95% CI = 1.033–2.679, P = 0.039). CTLA‐4 +49GG genotype frequency was found to be significantly lower in patients with BD than those of healthy controls (4% vs. 10.6%, OR = 0.357, 95% CI = 0.130–0.983, P = 0.05). Genotype and allele frequencies of the CTLA‐4–318C/T polymorphism between the BD and healthy control groups were not significantly different (12.2% vs. 10.6%, OR = 1.170, 95% CI = 0.570–2.402, P = 0.713). There were no associations between the studied polymorphisms and the main clinical features of BD. The frequencies of HLA‐B*51 were 60.3% and 30.7% in BD and control groups, respectively (OR = 3.429, 95% CI = 2.115–5.559, P = 0.0001). Association between HLA‐B*51 and each studied polymorphism did not reach to significant levels (OR = 0.479, 95% CI = 0.228–1.004, P = 0.064 for CD28 IVS3 +17TT genotype; OR = 2.180, 95% CI = 1.025–4.639, P = 0.061 for TC genotype; OR = 1.570, 95% CI = 0.870–2.836, P = 0.146 for C allele). These results may suggest that CD28 IVS3 +17TC genotype may be a risk factor for the development of BD, on the contrary CTLA‐4 +49GG genotype may be protective in the studied Turkish population.

Different clinical presentation of the hyperimmunoglobulin D syndrome (HIDS) (four cases from Turkey)

Hyperimmunoglobulin D syndrome (HIDS) is one of the autoinflammatory syndromes which are characterized by febrile attacks. Duration and frequency of the febrile attacks, as well as typical organ involvements vary greatly. Recently, it is possible to reach more reliable data by the possibilities that are opened up by molecular genetics in order to highlight the aetiopathogenesis of this group of diseases. Typical patients with HIDS have an onset of disease in the first year of life. Here, we report four Turkish HIDS cases; three of whom, the symptoms started at a later age. The diagnoses were made by relevant clinical symptoms along with MVK mutations detected by DNA sequencing method. As summarised in this article, HIDS could be presented with a broad spectrum of symptoms. Although most of the HIDS patients are reported from Europe and especially Dutch ancestry, case reports are presented from all over the world. For this reason, HIDS should be kept in mind for the differential diagnosis of periodic fever syndromes or before accepting an FMF patient as colchichine resistant. We suppose that the phenomenon of “later-onset HIDS” should shed light into unresolved clinical problems of patients with periodic fever. Especially in countries that FMF is more frequent such as Turkey, even though the symptoms start later than classic cases, HIDS should be kept in mind for differential diagnosis of periodic fever syndromes.

Role of KIR genes and genotypes in susceptibility to or protection against hepatitis B virus infection in a Turkish cohort

Background Killer cell immunoglobulin-like receptors (KIRs) are a family of inhibitory and activating receptors expressed by natural killer (NK) cells and regulate NK cell activity in the innate response against viral infections. The aim of this study was to determine the possibility of KIR genes and genotypes as a candidate for susceptibility to or protection against chronic hepatitis B virus (HBV) infection or spontaneous remission of the infection in a Turkish cohort. Material/Methods The present study was carried out on 37 patients with chronic HBV infection, 36 patients in spontaneous remission of HBV infection, and 85 healthy subjects. Sequence-specific oligonucleotide probes analysis was used to investigate 16 KIR genes. All data were statistically analyzed by the Fisher exact test. Results The rate of inhibitory KIR2DL3 (p=0.0) and 3DS1 (p=0.0) were higher in the healthy group than the group composed of chronic HBV patients and patients with spontaneous remission. There were no statistically significant differences between the rate of AA and Bx genotypes of chronic HBV patients and patients with spontaneous remission and the control group (p>0.05). Conclusions Our results suggest that KIR2DL3 and KIR3DS1 genes could be protector genes for HBV infection and they could be important immuno-genetic markers in determining antiviral immunity in the Turkish population.

Analysis of vascular endothelial growth factor gene 936 C/T polymorphism in patients with familial Mediterranean fever.

Vascular endothelial growth factor (VEGF) is a cytokine that promotes endothelial cell proliferation, leucocyte chemotaxis and expression of adhesion molecules and is a major mediator of vascular permeability. It has been demonstrated that VEGF directly activates neutrophils and it could promote acute recruitment of leucocytes. It is known that neutrophils are the major cell population involved in acute inflammation in familial Mediterranean fever (FMF) and the role of VEGF in these cells may be crucial. The aim of this study was to investigate whether the 936 C/T functional polymorphism of the VEGF gene is associated with susceptibility to FMF and its relationship with the main clinical features of the disease. Polymerase chain reaction–restriction fragment length polymorphism technique was used to determine 936 C/T polymorphism within the VEGF gene in 75 patients with FMF and 122 non‐related healthy controls. Genotype and allele frequencies of the VEGF 936 C/T polymorphism between patients with FMF and healthy control groups were not significantly different (OR = 0.74, 95% CI = 0.40–1.37, P = 0.335 for CT genotype; OR = 1.11, 95% CI = 0.67–1.83, P = 0.700, for T allele). Although VEGF 936 TT genotype was found to be more frequent in patients with FMF than in healthy controls (6.7% vs. 1.6%, respectively), the difference was not significant (OR = 4.28, 95% CI = 0.81–22.67, P = 0.108). No associations were found between the studied polymorphism and either the clinical features such as arthritis, abdominal pain, pleuritis, myalgia, arthralgia and erysipelas‐like erythema of the disease or the four common studied exon 10 mutations (M694V, M680I, V726A, M694I) of the Mediterranean fever gene. Present results suggest that VEGF gene 936 C/T polymorphism does not seem to be associated with susceptibility to FMF and its clinical manifestations.

Mevalonate kinase gene mutations and their clinical correlations in Behçet's disease

Genetics is suggested to play a role in the development of Behçets disease (BD). Shared phenotipic features requires an approach to differential diagnosis from periodic febrile syndromes. We planned to study for mevalonate kinase (MVK) as a candidate for a susceptibility gene for Behçets disease.

MANNOSE BINDING LECTIN (MBL) GENE POLYMORPHISMS AND THEIR RELATIONS WITH CLINICAL FEATURES IN PATIENTS WITH FAMILIAL MEDITERRANEAN FEVER (FMF)

Amac: Kompleman aktivasyonunun lektin yolaginda rol oynayan ve dogal immun sistemin bir parcasi olan mannoz baglayici lektin (MBL), cesitli patojenlerin mannan gruplarinin uyarisi ile aktive olur. MBL genindeki bazi polimorfizmler (orn. kodon 52, kodon 54 polimorfizmleri) MBL’nin serum duzeylerinde degisikliklere yol acarak, infeksiyon hastaliklarina yatkinliga neden olabildigi gibi, bazi otoimmun ve inflamatuvar hastaliklarin patogenezine de katkida bulunabilir. Bu calismada, ailesel Akdeniz atesi (AAA) hastalarinda MBL geni kodon 52 ve kodon 54 polimorfizmlerinin sikligini ve basta sekonder amiloidoz olmak uzere, hastaligin klinik ozellikleri ile olasi iliskilerini arastirmayi amacladik. Gerec ve Yontem: Yuzelli-yedi AAA hastasinda ve hastalarla akrabalik iliskisi olmayan benzer demografik dagilimdaki 150 saglikli kontrolde MBL genindeki R52C C>T ve G54D G>A polimorfizmleri sekanslama yontemi ile arastirildi. AAA hastalarinin MEFV geni analizleri, klinik ozellikleri ve ataksiz donemdeki serum CRP duzeyleri kaydedildi. Genetik sonuclar ile klinik ve laboratuvar bulgular arasindaki olasi iliskiler incelendi. Bulgular: MBL geni R52C C>T polimorfizmi hastalarin %12,7’sinde, kontrol grubunun %10,6’sinda saptandi. G54D G>A polimorfizmi hastalarin %26,8’sinde, kontrollerin %26,7’sinde saptandi. Polimorfizm sikligi acisindan, iki grup arasinda anlamli fark bulunamadi (p=0,79 ve 0,98). Incelenen MBL geni polimorfizmleri ile hastalarin cesitli klinik ozellikleri (or. amiloidoz, ates, kolsisin yaniti, MEFV mutasyonlari) arasinda anlamli iliski bulunamadi. AAA hastalarinin ortalama CRP degeri 4,90±6,72 mg/dL olup, ataksiz donemde serum CRP duzeyi normalden yuksek (>0,8 mg/dL) olan hastalarda MBL kodon 52 polimorfizmi sikligi %25,2, kodon 54 polimorfizmi sikligi %14,8 oraninda bulundu. AAA hastalarinda yuksek CRP duzeyine gore kodon 52 ve kodon 54 polimorfizmi sikliklari farkli bulunmadi (p=0,399). AAA hastalarinda M694V mutasyonu ile amiloidoz arasinda (p=0,002) ve M694I mutasyonu ile kolsisin direnci arasinda (p=0,016) anlamli iliski saptandi. Sonuc: AAA hastalarin ataksiz donemdeki CRP duzeyleri ve tasidiklari klinik ozellikler ile MBL polimorfizmleri arasinda anlamli iliski bulunamamasi, AAA hastalarinin proinflamatuvar durumda oldugunu ve MBL aracili mekanizmalarin bu sureclere katkisinin olmadigini dusundurmektedir. Olgularimizda M694I ile kolsisin direnci arasinda anlamli iliski saptanmis olmasi dikkate deger bir bulgudur. Yine olgularimizda M694V ile amiloidoz arasinda anlamli iliski bulunmasi onceki literatur bulgulari ile uyumludur ve M694V’nin hastalik siddeti ve prognozu icin onemli oldugu gorusunu desteklemektedir.

FRI0412 Mannose Binding Lectin Gene Polymorphisms in Spondyloarthropathies

Background Genetic and environmental factors are known to play a role in the pathogenesis of spondyloarthropathies (SPA). Mannose binding lectin (MBL) which is a member of collectin family proteins, is an important component of innate immunity. Low levels of MBL is associated with susceptibility to infections. Several variants of MBL gene may result in MBL deficiency. MBL gene polymorphisms and MBL deficiency are shown to be related with several autoimmune and inflammatory diseases. Objectives The relationship between the MBL levels or MBL gene polymorphisms and SPA predisposition has previously been investigated in a few small studies with conflicting results. In this particular study, we aimed to assess the relationship of the MBL genes in the develepoment of SPA in a group of Turkish patients. Methods Study included 132 patients with SPA (75 female, 57 male, mean age: 43.0±12.4) and 87 healthy subjects (58 women, 29 men, mean age: 40.3±13.4) as control group. Eighty nine of the SPA patients had ankylosing spondylitis (AS) and 43 had non-AS SPA. MBL gene single nucleotide polymorphisms (SNP) in the codons 52, 54 and 57 of exon 1 were studied by sequencing method. Results G54D G>A polymorphism was not different between the patient and the healthy control groups (p=0.291). The frequencies of R52C C>T and G57E G>A polymorphisms were found to be significantly higher in the SPA patients than those of the healthy controls (p=0.019 and 0.001 respectively). While the non-AS subgroup showed significancy for both R52C C>T and G57E G>A polymorphisms (p=0.004 and 0.000 respectively), the AS subgroup showed significancy only for G57E G>A (p=0.113 and 0.014 respectively). Clinical characteristics of either AS or non-AS SPA patients were not associated with any of the MBL polymorphisms. In a previous study, haplotypes of MBL genetic polymorphisms were found to be associated with AS in Korean patients. Our study is the first one from Turkey indicating the relationship between SPAs and MBL gene polymorphisms. Conclusions Even though we had rather small sample size, our results showed significantly higher R52C C>T and G57E G>A polymorphisms in Turkish patients with SPA. Therefore we may suggest that MBL gene polymorphisms could be related with the predisposition of the disease. References Churl Hyun Im et al.: Mannose-binding lectin 2 gene haplotype analysis in Korean patients with ankylosing spondylitis. Rheumatol Int (2012) 32:2251–5 Aydin SZ et al: Mannose binding lectin levels in spondyloarthropathies. J Rheumatol. (2007) Oct;34(10):2075–7 Aydin SZ et al: Mannose binding lectin levels are not related to radiographic damage in ankylosing spondylitis Rheumatol Int (2010) 30:415–7 Disclosure of Interest None declared

Killer Cell Immunoglobulin-Like Receptor (KIR) Genotype Distribution in Familial Mediterranean Fever (FMF) Patients

Background Familial Mediterranean fever (FMF) is an autosomal recessive autoinflammatory disease predominantly affecting Mediterranean populations. The gene associated with FMF is the MEFV gene, which encodes for a protein called pyrin. Mutations of pyrin lead to uncontrolled attacks of inflammation, and subclinical inflammation continues during attack-free intervals. Killer cell immunoglobulin-like receptor (KIR) genes encode HLA class I receptors expressed by NK cells. The aim this study was to look for immunogenetic determinants in the pathogenesis of FMF and find out if KIR are related to susceptibility to disease or complications like renal amyloidosis. Material/Methods One hundred and five patients with FMF and 100 healthy individuals were involved in the study. Isolated DNA from peripheral blood was amplified by sequence specific PCR probes and analyzed by Luminex for KIR genotypes. Fisher Exact test was used to evaluate the variation of KIR gene distribution. Results All patients and healthy controls expressed the framework genes. An activator KIR gene, KIR2DS2, was significantly more frequent in FMF patients (p=0.036). Renal amyloidosis and presence of arthritis were not associated with KIR genes and genotype. KIR3DL1 gene was more common in patients with high serum CRP (p=0.016). Conclusions According to our findings, we suggest that presence of KIR2DS2, which is an activator gene for NK cell functions, might be related to the autoinflammation in FMF. The potential effect of KIR genes on amyloidosis and other clinical features requires studies with larger sample sizes.

AB0151 Distribution of Natural Killer Cell Immunoglobulin-Like Receptor (KIR) Genes in Turkish Patients with Ankylosing Spondylitis

Background Genetic and environmental factors are known to play a role in the pathogenesis of ankylosing spondylitis (AS) which is the most severe subgroup of patients of spondyloarthritis. Killer Cell Immunoglobulin-like receptors (KIR) are the molecules regulating the cytotoxic response of Naturel Killer (NK) cells. KIR genes have polymorphic structures and comprise a high difference between the populations. KIR genes have been shown to be associated with the pathogenesis of several autoimmune and inflammatory diseases. Objectives The relationship between the KIR genes and AS predisposition was already shown in previous studies in different populations. In this particular study, we aimed to assess the relationship of the KIR genes in the develepoment of AS in a group of Turkish patients. Methods Study includes 68 patients with AS (31 female, 37 male, mean age: 38.27) and 75 healthy subjects (34 women, 41 men, mean age: 38.6) as a control group. Typing of sixteen different KIR genes was performed by the method of Spesific Sequence Oligonucleotide Probes (SSOP). The distribution of KIR genotypes were obtained from the data base of “www.allelefrequencies.net”. The rates of the KIR genes of the both groups were found by direct counting method and the comparision of the groups is made by Fishers Exact test. Results The frame genes KIR2DL4, 3DL2, 3DL3 and the 3DP1 were present in all patients and controls. KIR3DS1 which is one of the activating KIR genes, was found to be significantly higher in the AS cases (39.7%) than that of the control group (16%) (p=0.002), (Figure 1). In addition, there was no significant difference between AA and Bx genotypes in the patient and control groups. In the meta-analysis published by Zuo and colleagues in 2014; seven different studies conducted in Iran, Chinese, Spanish, English, Russian and Polish populations between the years of 2009-2012 were examined and it was reported that several KIR genes such as KIR2DL1, KIR2DS4, KIR2DS5 and KIR3DS1 were found to be associated with AS susceptibility. This is the first study from Turkey about the relationship between AS and KIR genes. We found KIR3DS1 gene significantly higher in Turkish AS patients as similar to the results of this meta-analysis study. Conclusions These results show that KIR3DS1 gene activating NK cell cytotoxicity may trigger AS development and suggesting that it is related with the disease predisposition. Our result is compatible with other Caucasian populations. References Yu-Lian Jiao et al.: Polymorphisms of KIRs Gene and HLA-C Alleles in Patients with Ankylosing Spondylitis: Possible Association with Susceptibility to the Disease. J Clin Immunol (2008) 28:343–349 Hai-Ning Zuo et al.: Genetic variations in the KIR gene family may contribute to susceptibility to ankylosing spondylitis: a meta-analysis, Mol Biol Rep (2014) 41:5311–5319 Disclosure of Interest None declared

P02-001 – A novel TNFRSF1A mutation in periodic fever

Tumor necrosis factor receptor-associated periodic syndrome (TRAPS) is an autosomal dominant autoinflammatory disease characterized by periodic fever, accompanying with attacks of abdominal pain, arthralgia, myalgia, erythematous rashes, periorbital edema and conjunctivitis. Mutations in the extracellular domain of the 55-kD tumor necrosis factor receptor (TNFRSF1A) has been shown to be responsible for the TRAPS syndrome.