An Alternative Perspective to the FMF Clinic: MCP-1 (A-2518G) and CCR2 (G190A) Polymorphisms and MCP1 Expression

Abstract

Background: Familial Mediterranean Fever (FMF) is an autoinflammatory disease and may express as various clinical findings. Chemokines are crucial elements of the inflammatory process. MCP-1 and its’ receptor CCR2 are the main chemokines for monocytes/macrophages that may play critical roles in FMF. Thus, it was aimed to investigate the MCP-1 (A-2518G) and CCR2 (G190A) polymorphisms and MCP-1 expression level, which may affect MEFV gene function. Material and Method: Patients with FMF were identified according to the Tel-Hashomer criteria. DNA and RNA were isolated from the obtained blood samples. Genotyping analysis was performed by PCR-RFLP technique. In addition, expression analyzes were performed by Real-time PCR method. The obtained results were evaluated statistically. Results: A total of 229 individuals (125 male and 104 female) were included in the study. While 120 individuals had FMF clinic, and 107 individuals did not have. The remaining two individuals had suspicious clinical status. In addition, while 75 individuals were homozygous mutants, 77 individuals were heterozygous mutants, and 77 individuals did not carry mutation in the MEFV gene. No significant relationship was found in between both FMF clinic and MEFV genotypes, and MCP-1 (A-2518G) and CCR2 (G190A) genotypes. In the expression analysis, MCP-1 expression increased in patients with FMF clinic compared to those without. In addition, MCP-1 expression was increased in the heterozygous MEFV group compared to those without mutation, moreover, the expression level was highest in homozygous MEFV group. In addition, according to the MCP-1 (A-2518G) genotyping, MCP-1 expression elevated in the homozygous as well as the heterozygous groups, compared to the Wild type group. Conclusion: MCP-1 expression is increased in FMF disease, which may explain the clinical differences between FMF patients. MEFV mutations may exacerbate inflammation by increasing MCP-1 transcription. MCP-1 expression is increased in patients with MCP-1(A-2518G) mutations, which aggravates FMF clinic. MCP-1 expression may be assessed as a marker in suspicious cases. Keywords: Familial Mediterranean Fever, MCP-1, CCR2, expression

Keywords

• Familial Mediterranean Fever
• MCP-1
• CCR2
• expression

FMF Kliniğine Alternatif Bir Bakış Açısı: MCP-1 (A-2518G) ve CCR2 (G190A) Polimorfizmleri ve MCP1 Ekspresyonu

Abstract

Amaç: Ailevi Akdeniz Ateşi (AAA) otoinflamatuar bir hastalıktır ve çeşitli klinik bulgular olarak kendini gösterebilir. Kemokinler, inflamatuar sürecin önemli unsurlarıdır. MCP-1 ve onun reseptörü CCR2, FMF'de kritik roller oynayabilen monositler/makrofajlar için ana kemokinlerdir. Bundan dolayı MEFV gen fonksiyonunu etkileyebilecek MCP-1 (A-2518G) ve CCR2 (G190A) polimorfizmlerinin ve MCP-1 ekspresyon düzeyinin araştırılması amaçlanmıştır. Gereç ve Yöntem: FMF'li hastalar Tel-Hashomer kriterlerine göre belirlendi. Elde edilen kan örneklerinden DNA ve RNA izole edildi. Genotipleme analizi, PCR-RFLP tekniği ile yapıldı. Ayrıca Real-time PCR yöntemi ile ekspresyon analizleri yapıldı. Elde edilen sonuçlar istatistiksel olarak değerlendirildi. Bulgular: Çalışmaya toplam 229 birey (125 erkek ve 104 kadın) dahil edildi. Bunlardan 120 kişide FMF kliniği bulunurken, 107 kişide yoktu. Kalan iki kişi şüpheli klinik duruma sahipti. Çalışmaya alınan bireyler MEFV genotiplemesine göre değerlendirildiğinde ise 75 birey homozigot mutant, 77 birey Heterozigot saptanırken 77 birey ise MEFV geninde mutasyon taşımıyordu. Yapılan analizde Hem FMF kliniği hem de MEFV genotipleri ile MCP-1 (A-2518G) ve CCR2 (G190A) genotipleri arasında anlamlı bir ilişki bulunmadı. Ekspresyon analizinde, FMF kliniği olan hastalarda olmayanlara göre MCP-1 ekspresyonu artmış olarak saptandı. Ayrıca heterozigot MEFV grubunda mutasyonu olmayanlara göre MCP-1 ekspresyonu artmış olarak saptandı, Dahası homozigot MEFV grubunda MCP-1 ekspresyonu en yüksek düzeydeydi. Ek olarak, MCP-1 (A-2518G) genotiplendirmesine göre, MCP-1 ekspresyonu, Wild type gruba kıyasla hem homozigot hem de heterozigot gruplarda yükselmiştir. Sonuç: FMF hastalığında MCP-1 ekspresyonu artmış olup, bu durum FMF hastaları arasındaki klinik farklılıkları açıklayabilir. MEFV mutasyonları, MCP-1 transkripsiyonunu artırarak inflamasyonu şiddetlendirebilir. MCP-1(A-2518G) mutasyonlu hastalarda MCP-1 ekspresyonu artar, bu da FMF kliniğini ağırlaştırır. Anahtar Kelimeler: Ailevi Akdeniz Ateşi, MCP-1, CCR2, Expresyon analizi

Keywords

• Ailevi Akdeniz Ateşi
• MCP-1
• CCR2
• Expresyon analizi

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