The Effect of Storage Conditions of Erythrocyte Suspensions on T Cell Viability and Proliferation

Abstract

Transfusion-related immunomodulation (TRIM) can be defined as changes in the immune system of the recipient caused by allogeneic blood transfusion (ABT). The most commonly blamed agent is mononuclear (MNC) cells in the blood component. Therefore, T cells (CD3+CD4+ and CD3+CD8+) in erythrocyte suspensions (ES) were targeted in our study. The effect of storage conditions on viability, proliferation, and activation levels were analysed. ESs were obtained from whole bloods donated from three donors. Whole blood sample (5th hour) and ES samples (0, 7, 14, 21, 42nd day) were achieved from each blood component. Two samples taken from donors in EDTA tubes before donation were also included in the study. MNCs were isolated from these samples and analyses were performed with these cells by flow-cytometer. T-cell viability disappeared in the 42nd-day ES samples, whereas proliferation capacity was lost in the 21st-day ES samples. Activation marker levels increased at 16 and 72 hours compared to 0 hours MNH culture. Also, distinct differences were observed among results according to the ages of the donors. In conclusion, the viability and proliferation capacity of T lymphocytes in the ES decreases with the effect of storage time and conditions. These results suggest that the relationship between allogeneic T lymphocytes and the development of TRIM may be low; the increased activation capacity of T lymphocytes when they are removed from ES suggests that erythrocytes may show suppressive properties; and demographic parameters such as donor and patient age may also play a role in the development of TRIM.

Keywords

• Erythrocyte suspension
• viability
• proliferation
• immunomodulation
• T lymphocyte

Eritrosit Süspansiyonlarının Depolanma Koşullarının T Hücre Canlılığı ve Proliferasyonu Üzerindeki Etkisi

Abstract

Önemli transfüzyon komplikasyonlardan biri olan transfüzyonla ilişkili immün düzenlenme (TRIM), allojeneik kan transfüzyonunun (AKT) alıcının immün sisteminde yol açtığı değişiklikler olarak tanımlanabilir. En çok suçlanan etken, kan bileşeni içindeki mononükleer (MNH) hücrelerdir. Bu nedenle çalışmamızda eritrosit süspansiyonları (ES) içindeki T hücreler (CD3+CD4+ ve CD3+CD8+) hedeflenmiş, depolama koşullarının etkisiyle canlılık, proliferasyon ve aktivasyon düzeylerindeki değişimler incelenmiştir. Bu amaçla, üç adet kan bağışçısından alınan tam kanlardan ES’ler elde edilmiştir. Her kan bileşeninden tam kan örneği (5. saat) ve ES örnekleri (0, 7, 14, 21, 42. gün) elde edilmiştir. Ayrıca bağışçıdan bağış öncesi EDTA’lı tüplere alınan iki adet örnek de çalışmaya katılmıştır. Analizler bu örneklerden ayrıştırılan MNH kullanılarak yapılmıştır. Canlılık analizleri doğrudan MNH’ler, proliferasyon ve aktivasyon analizleri MNH kültürleri aracılığıyla akan hücre ölçerde gerçekleştirilmiştir. Canlılık düzeylerinin depolama süresi ortalarında azalmaya başladığı, 42. gün ES örneklerinde hemen tamamen yok olduğu belirlenmiştir. T hücrelerin proliferasyon becerisi daha erken azalmış ve 21. gün ES örneklerinde kaybolmuştur. Aktivasyon belirteci düzeyleri MNH kültürünün sıfırıncı saatlerine göre 16 ve 72 saatlerde artış göstermiştir. Ayrıca bağışçıların yaşlarına göre de sonuçlarda belirgin farklılıklar gözlemlenmiştir. Sonuç olarak ES depolama süresi ve koşullarının etkisiyle ürün içindeki T lenfositlerin canlılığı ve proliferasyon becerileri azalmaktadır. Bu sonuçlar allojeneik T lenfositlerin TRIM gelişimiyle ilişkilerinin düşük olabileceğini; T lenfosit aktivasyon kapasitelerinin ES’den uzaklaştıklarında artmış göstermesi eritrositlerin baskılayıcı özellik gösterebildiğini; TRIM gelişiminde bağışçı ve hasta yaşı gibi demografik parametrelerin de rol oynayabileceğini düşündürmektedir.

Keywords

• Eritrosit süspansiyonu
• canlılık
• proliferasyon
• immün düzenlenme
• T lenfosit

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