Sporadik behçet sendromlu bireylerde hla-b geninin ekspresyon analizi

Behçet's Syndrome (B) is a mutisystemic, rheumatologic inflammatory disease characterized by primary symptoms of mucocutaneous lesions, vasculitis, oral and genital ulcerations. BS, individuals have been violently affecting their lives for two or three decades. It is mostly seen in the Mediterranean and the Middle East. The frequency of 100,000 population ranged from 20 to 420 in Turkey. The HLA-B51 allele, an allele of the HLA-B (human leukocyte antigen) gene, is one of the most important determinants associated with BS to date. Although the etiology of BS is not completely known, it emphasizes the importance of genetics in the etiology of the syndrome a strong linking between HLA-B51 and BS. The task of HLA-B is to induce antigen presentation to T cells in the immune system. In current genomic associative studies, different genes such as IL-1, MICA, STAT4 GIMAP2 / GIMAP4 have been found to be associated with BS. As a basis to the this study, in our group a global methylation study was conducted to understand the effects of epigenetic mechanisms. However, at the global level, the expected 5-mdC: dC ratio in the genome between both Behçet twins and healthy control twins was between 0.02 and 0.05. As a result, there was no difference between the twins with Behçet's syndrome and the healthy twins and it was thought that global methylation might not be effective in the pathology of the disease. Later on it was aimed to investigate whether the methylation profile of the HLA-B51 allele differs between the twins with Behçet's syndrome and the healthy twins, considering that the allele-specific methylation may play a role in the pathology of the disease. The current bioinformatics studies indicate a CpG island that starts from the promoter region of the HLA-B51 antigen and extends to the middle of the third intron. It was decided to investigate the presence of the methylation levels of the HLA-B gene in the promoter, 1st exon, 1st intron, 2nd exon, 2nd intron and 3rd exon regions of the HLA-B gene and the etiology of Behçet's disease. It is revealed that 4 monozygotic and 4 dizygotic Behçet's syndrome twins have a epigenetic regulation of 1st and 2nd exons of HLA-B gene as a result of this study. After this study, it is planned to better understand the genetic and epigenetic effects of the HLA-B51 allele on cases of familial Behçet syndrome and other Behçet syndrome individuals and their healthy family members in their families. Rates of methylation in all BS individuals were found to be significantly higher than those of healthy relatives. However, when all groups were examined separately, it was found that HLA-B51 carriage did not make a significant difference on the methylation levels. The results we obtained were consistent with the results of our previous work with Behçet twins. In the guidance with this knowledge , possible epigenetic mechanisms independently of the HLA-B51 allele are thought to be effective in the development of the disease. In this study HLA-B51 allele carriage, expression level of HLA-B gene and promotor methylation of HLA-B gene were investigated in individuals with sporadic Behçet's syndrome and healthy individuals. For this purpose, individual BS patients were randomly selected and the individual was asked whether or not he or she was only BS among his / her family and relatives.According to this criterion, 46 patients with BS were selected. To determine the HLA-B51 carriage status of these patients, sequence-specific PCR (Polymerase chain reaction) followed by enzyme cleavage reaction was performed. RT-QPCR method was used to measure the transcript levels of HLA-B in BS patients classified by this method. The delta Ct values obtained by RT-QPCR were compared between groups using the t test for statistical analysis. 6 patients with high HLA-B expression with HLA-B51 and 6 patients with low HLA-B51 expression and 5 healthy controls with HLA-B51 and high expression level and those with no HLA-B51 expression and low HLA-B was measured using the One Step Q-methyl Kit based on Real-Time PCR to see the methylation differences of the healthy control promotor. 17 healthy controls for comparison with BS patients were included in the study. In this study, 34 sporadic BS individuals were found to be HLA-B51 positive and 12 sporadic BS individuals were found to be HLA-B51 negative. In healthy controls, 11 patients were found to be HLA-B51 positive and 6 individuals were found to be HLA-51 negative. From the results of statistical analyses, transcript levels compared between different combinations of groups. HLA-B transcript level is found to be higher in BS patients when compared to healthy control group (p= 0.0391).Promotermethylationlevel of HLA-B is foundto be lower in BS patientswhoarepositivefor HLA-B51 when compared to BS patients who are negative for HLA-B51 (p=0.02080). Also in thiscomparisongrouptranscriptlevel of HLA-B andpromotermethylationlevel of HLA- B showed a negativecorrelation (r= -0.8944) as expected.