Adenosine deaminase mediates endothelial inflammation via an ADA1-CD26 interaction in post-COVID

Ada Kawecka¹Klaudia Stawarska¹Marzena Romanowska-Kocejko²Marta Żarczyńska-Buchowiecka²Agata Jedrzejewska¹Alicja Braczko¹Milena Deptuła³Malgorzata Zawrzykraj⁴Oliwia Król¹Marika Frańczak¹Gabriela Harasim¹Michał Pikuła³Marcin Hellmann²Barbara Kutryb-Zajac^1*

• ¹Department of Biochemistry, Medical University of Gdansk, Gdańsk, Pomeranian, Poland
• ²Department of Cardiac Diagnostics, Medical University of Gdansk, Gdansk, Poland
• ³Laboratory of Tissue Engineering and Regenerative Medicine, Medical University of Gdansk, Gdansk, Pomeranian, Poland
• ⁴Division of Clinical Anatomy, Department of Anatomy, Medical University of Gdansk, Gdansk, Poland

Adenosine deaminase (ADA) isoenzymes play a role in microvascular dysfunction following SARS-CoV-2 infection. This study analyzes the mechanisms behind ADA1-dependent endothelial inflammation in post-COVID-19 syndrome. We investigated whether immune cells from post-COVID patients could contribute to the increased total ADA activity. Additionally, we examined ADA's enzymatic and extra-enzymatic activities in human primary lung microvascular endothelial cells (HULECs) stimulated with post-COVID patients' serum.Treatment of HULECs with sera from post-COVID patients resulted in elevated levels of the ADA1 isoenzyme and the ADA1-anchoring protein, CD26. This increase correlated with enhanced adhesion of THP-1 monocytes/macrophages to HULECs. Inhibiting the ADA1-CD26 interaction with glycoprotein-120 prevented the rise in cell-surface ADA levels in HULECs and reduced the adhesion of THP-1 cells to the endothelium. A similar effect was observed when HULECs were pre-incubated with the SARS-CoV-2 spike protein, which co-localized with CD26 in activated HULECs.We propose that ADA1 may promote vascular inflammation in post-COVID-19 syndrome through both canonical and non-canonical mechanisms. On one hand, its increased enzymatic activity can suppress adenosine-dependent pathways. On the other hand, ADA1 may function as an adhesion molecule facilitating interactions between immune cells and the endothelium via ADA1-CD26 complexes.