ADA2 enzim düzeyleri ve aktivitesinin, COVİD-19 enfeksiyon şiddeti üzerindeki etkisi
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Date
2023-07-13
Authors
Bozkurt, Tuğçe
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
COVID-19; Aralık 2019'da Çin/Wuhan'da ortaya çıkan tüm dünyayı etkisi altına alan pandemik hastalıktır. SARS-CoV-2, bulaşıcılığı yüksek, ağır klinik tablolara sebep olabilen enfeksiyon etkenidir. Adenozin deaminaz (ADA) özellikle lenfoid dokularda bulunan, immün sistem fonksiyonlarını etkileyebilen, ADA1 ve ADA2 (CECR1) izoenzimlerine sahip bir enzimdir. ADA1 T hücre proliferasyonunu, ADA2 ise monositlerin farklılaşmasını etkileyebilmektedir. Çalışmamızda; 41 ağır vaka, 34 hafif/orta vaka, 39 a/presemptomatik ve 6 kritik seviye yetişkin COVID-19 hastası ve 40 sağlıklı kontrol grubu dahil edilmiştir. Enzim aktivitesi Giusti ve Galanti yöntemi ile çalışılmıştır. 6 sağlıklı kontrolden elde edilen PKMH ve monositler ile eş-kültür deneyleri yapılmıştır. Eş-kültür sonrası elde edilen süpernatanlar ve COVID-19 hasta serumları, ELISA yöntemi ile incelenerek M1 ve M2 ilişkili sitokin/kemokin profilleri ortaya konmuştur. ADA enzim aktiviteleri incelendiğinde en yüksek değeri kritik COVID-19 vakaları oluşturmaktadır. Sitokin/kemokin çalışmalarında; IL-6, TNF-α, IL-12, IFN-ɣ, GM-CSF, ROS, iNOS, arjinaz, IL-4, IL-10 ve M-CSF'de kritik ve ağır vakalarda artış gözlemledik. Hücre kültürü deneylerimizde ise; kritik, ağır vakalar ve in vitro COVID-19 grubunda, ADA2'in M2 ilişkili sitokin ve kemokinlerle pozitif korelasyon gösterdiği bulunmuştur. Bunun yanında, AHÖ incelemelerinde CD206+ CD86+ M1-M2 miks makrofajlar COVID-19 grubunda anlamlı derecede yüksek bulunmuştur. Sonuç olarak; makrofaj polarizasyonunun ADA2 ile doğrudan bağlantılı olduğu, ADA1'in ise daha düşük katkı sağladığına dair bulgular elde edilmiştir. Tüm analizlere göre, ağır COVID-19 vakalarında makrofajların M1'den M2'e yönelik bir polarizasyon gösterdiği, fakat daha çok M1-M2 karışık fenotipte heterojenik bir yapı göstermekte olduğuna dair kanıtlar elde edilmiştir. Bu durum, COVID-19'da, ADA2'in makrofaj polarizasyonuna etkisini ve hastalık süreciyle muhtemel ilişkisini gösterebilir.
COVID-19 is a pandemic disease that emerged in December 2019 in China/Wuhan and has affected the whole world. SARS-CoV-2 is the infectious agent responsible for severe clinical manifestations. Adenosine deaminase (ADA) is an enzyme primarily found in lymphoid tissues and can affect immune system functions. It has two isoforms, ADA1 and ADA2 (CECR1), which can influence T-cell proliferation and monocyte differentiation, respectively. In our study, we included 41 severe cases, 34 mild/moderate cases, 39 asymptomatic/pre-symptomatic cases, and 6 critical adult COVID-19 patients, as well as 40 healthy individuals as the control group. The enzyme activity was measured using the Giusti and Galanti method. We performed co-culture experiments using peripheral blood mononuclear cells (PBMCs) and monocytes isolated from 6 healthy controls. The supernatants obtained after co-culture and COVID-19 patient sera were analyzed using the ELISA method to determine the cytokine/chemokine profiles associated with M1 and M2 phenotypes. When examining ADA enzyme activities, the highest values were observed in critical COVID-19 cases. In cytokine/chemokine studies, we observed an increase in IL-6, TNF-α, IL-12, IFN-ɣ, GM-CSF, ROS, iNOS, arginase, IL-4, IL-10, and M-CSF in critical and severe cases. In our cell culture experiments, we found that ADA2 exhibited a positive correlation with M2-associated cytokines and chemokines in critical, severe cases, and the in vitro COVID-19 group. Additionally, in flow cytometric analyses, CD206+ CD86+ M1-M2 mix macrophages were significantly higher in the COVID-19 group. In conclusion, our findings suggest a direct association between macrophage polarization and ADA2, with ADA1 playing a lesser role. Based on all analyses, macrophages in severe COVID-19 cases exhibit a polarization from M1 to M2, but they also exhibit a mixed phenotype of M1 and M2, indicating heterogeneity. This implies a potential relationship between ADA2, macrophage polarization, and the disease process in COVID-19.
COVID-19 is a pandemic disease that emerged in December 2019 in China/Wuhan and has affected the whole world. SARS-CoV-2 is the infectious agent responsible for severe clinical manifestations. Adenosine deaminase (ADA) is an enzyme primarily found in lymphoid tissues and can affect immune system functions. It has two isoforms, ADA1 and ADA2 (CECR1), which can influence T-cell proliferation and monocyte differentiation, respectively. In our study, we included 41 severe cases, 34 mild/moderate cases, 39 asymptomatic/pre-symptomatic cases, and 6 critical adult COVID-19 patients, as well as 40 healthy individuals as the control group. The enzyme activity was measured using the Giusti and Galanti method. We performed co-culture experiments using peripheral blood mononuclear cells (PBMCs) and monocytes isolated from 6 healthy controls. The supernatants obtained after co-culture and COVID-19 patient sera were analyzed using the ELISA method to determine the cytokine/chemokine profiles associated with M1 and M2 phenotypes. When examining ADA enzyme activities, the highest values were observed in critical COVID-19 cases. In cytokine/chemokine studies, we observed an increase in IL-6, TNF-α, IL-12, IFN-ɣ, GM-CSF, ROS, iNOS, arginase, IL-4, IL-10, and M-CSF in critical and severe cases. In our cell culture experiments, we found that ADA2 exhibited a positive correlation with M2-associated cytokines and chemokines in critical, severe cases, and the in vitro COVID-19 group. Additionally, in flow cytometric analyses, CD206+ CD86+ M1-M2 mix macrophages were significantly higher in the COVID-19 group. In conclusion, our findings suggest a direct association between macrophage polarization and ADA2, with ADA1 playing a lesser role. Based on all analyses, macrophages in severe COVID-19 cases exhibit a polarization from M1 to M2, but they also exhibit a mixed phenotype of M1 and M2, indicating heterogeneity. This implies a potential relationship between ADA2, macrophage polarization, and the disease process in COVID-19.
Description
Keywords
Adenozin deaminaz, ADA1, ADA2, COVID-19, Adenosine deaminase
Citation
Bozkurt, T. (2023). ADA2 enzim düzeyleri ve aktivitesinin, COVID-19 enfeksiyon şiddeti üzerindeki etkisi. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.