Böbrek nakli hastalarında CD8+ T hücrelerindeki PD-1 ve TIM-3 ekspresyon düzeylerinin belirlenmesi

Abstract

Amaç: T hücresi immünoglobulin ve müsin domain 3 (TIM-3) ve programlanmış ölüm 1 (PD-1), T hücre yanıtının düzenlenmesinde önemli roller oynar. Böbrek naklinden sonra CD8+ T hücreleri böbreğe sızarak nekroz, tübülit ve hatta nakil reddine neden olabilir. Sonuç olarak böbrek nakli alıcılarından alınan periferik kan örneklerinde PD-1 ve TIM-3 genlerinin mRNA düzeylerini inceliyoruz. Gereç ve Yöntem: 29 hastadan hem transplantasyon öncesi (tx öncesi) hem de transplantasyon sonrası (tx sonrası) 58 periferik kan örneği toplandı. CD8+ T hücrelerini ayırmak için hemen MACS hücre ayırma sistemi kullanıldı. Toplanan hücrelerin saflığı flow sitometri ile belirlendi. RNA izolasyonu ve cDNA dönüşümünden sonra PD-1 ve TIM-3 genlerinin ifadelerini ölçmek için gerçek zamanlı polimeraz zincir reaksiyonu gerçekleştirildi. Bulgular: Hastaların %31, %45 ve %7'sinde PD-1 seviyelerinde artış, azalma ve değişiklik görülmezken, %76'sında Tim-3 seviyelerinde azalma, %21'inde ise artış görüldü. Ayrıca TIM-3 gen ekspresyon düzeylerinin rejeksiyon ataklarıyla pozitif yönde ilişkili olduğu belirlendi (p<0,001). Sonuç: Böbrek nakli öncesi ve sonrası TIM-3 mRNA düzeylerinin karşılaştırılması, hastaların klinik durumunun değerlendirilmesinde yararlı bir araç olabilir.

Keywords

• PD-1
• TIM-3
• Bağışıklık kontrol noktası düzenleyicileri
• Böbrek nakli

Evaluation of PD-1 And TIM-3 Expression Levels of CD8+ T Cells in Renal Transplant Patients

Abstract

Objective: After kidney transplantation, CD8+ T cells can infiltrate the kidney and cause necrosis, tubulitis, and even transplant rejection. For this reason, control of the T cell response is very important, and T cell immunoglobulin and mucin domain 3 (TIM-3) and programmed death 1 (PD-1) molecules play a role in regulating the T cell response. It is thought that the levels of TIM-3 and PD-1 expressions may be guiding in determining the clinical course after transplantation. This study aimed to determine the relationship between the mRNA levels of PD-1 and TIM-3 genes in peripheral blood samples taken from kidney transplant patients and the clinical conditions of the patients. Material and Method: 60 peripheral blood samples were collected from 30 kidney transplant patients, both pre-transplantation (pre-tx) and post-transplantation (post-tx). CD8+ T cells were separated from other lymphocytes by magnetic cell separation system (MACS) and their purity was determined by flow cytometry. Then, RNA was isolated and after cDNA conversion, the expressions of PD-1 and TIM-3 genes were determined by real-time polymerase chain reaction. Results: While it was determined that the TIM-3 gene expression level increased in patients with acute tubular necrosis, antibody-mediated rejection and cell-mediated rejection findings (p<0.001), no correlation was found between PD-1 expression levels and the clinical findings of the patients. Conclusion: It is thought that comparing TIM-3 mRNA levels before and after kidney transplantation may be a useful tool in evaluating the clinical status of patients.

Keywords

• PD-1
• TIM-3
• Immunecheckpoint regulators
• Renal transplantation

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