Augmented thrombin formation is related to circulating levels of extracellular vesicles exposing tissue factor and citrullinated histone-3 in anti-neutrophil cytoplasmic antibody-associated vasculitides

Asta Dogg Jonasdottir; Milena Manojlovic; Jelena Vojinovic; Annica Nordin; Annette Bruchfeld; Iva Gunnarsson; Fariborz Mobarrez; Aleksandra Antovic

doi:10.3389/fmed.2023.1240325

Augmented thrombin formation is related to circulating levels of extracellular vesicles exposing tissue factor and citrullinated histone-3 in anti-neutrophil cytoplasmic antibody-associated vasculitides

Front Med (Lausanne). 2023 Oct 17:10:1240325. doi: 10.3389/fmed.2023.1240325. eCollection 2023.

Authors

Asta Dogg Jonasdottir^{1

2}, Milena Manojlovic³, Jelena Vojinovic³, Annica Nordin⁴, Annette Bruchfeld^{1

5}, Iva Gunnarsson^{4

6}, Fariborz Mobarrez⁷, Aleksandra Antovic^{4

6}

Affiliations

¹ Department of Clinical Science, Intervention and Technology, Division of Renal Medicine, Karolinska Institutet, Stockholm, Sweden.
² Division of Nephrology, Department of Medicine, Landspitali - The National University Hospital, Reykjavik, Iceland.
³ Department of Pediatrics, Medical Faculty, University of Niš, Niš, Serbia.
⁴ Department of Medicine, Division of Rheumatology, Karolinska Institutet, Stockholm, Sweden.
⁵ Department of Health, Medicine and Caring Sciences, Linköping University, Linköping, Sweden.
⁶ Rheumatology, Karolinska University Hospital Stockholm, Stockholm, Sweden.
⁷ Department of Medical Sciences, Uppsala University, Uppsala, Sweden.

Abstract

Objectives: To study circulating myeloperoxidase (MPO)-positive extracellular vesicles (MPO⁺EVs) exposing citrullinated histone-3 (H3Cit), tissue factor (TF), and plasminogen (Plg) in association to thrombin generation in patients with anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitis (AAV).

Methods: We have involved well-characterized patients with AAV together with population-based controls. Flow cytometry was used to assess the levels of MPO⁺EVs in citrated plasma. MPO⁺EVs were phenotyped by anti-MPO-antibodies together with anti-CD142 (anti-TF), anti-H3Cit, and anti-Plg antibodies. A modified Calibrated Automated Thrombogram (CAT) assay was utilized to measure thrombin generation in plasma initiated by EVs-enriched pellets. The activity of AAV was evaluated with the Birmingham Vasculitis Activity Score (BVAS).

Results: This study comprised 46 AAV patients, 23 in the active stage of the disease and 23 in remission, as well as 23 age- and sex matched population-based controls. Augmented levels of all investigated MPO⁺ EVs were found in active AAV patients in comparison to the subgroup of patients in remission and controls. Thrombin generation, measured by endogenous thrombin potential (ETP) and peak of thrombin formation, was higher in plasma when triggered by EVs-enriched pellet from AAV patients. ETP and peak were associated with the levels of MPO⁺TF⁺ and MPO⁺H3Cit⁺ EVs. Additionally, MPO⁺TF⁺ EVs correlated with the disease activity evaluated with BVAS.

Conclusion: Augmented thrombin generation is found in AAV patients regardless of disease activity and is associated with higher exposure of TF and H3Cit on MPO⁺EVs. This may contribute to the increased risk of thrombosis seen in AAV patients.

Keywords: NETs; anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis; extracellular vesicles; thrombin generation; tissue factor.

Grants and funding

The study was performed by grants provided by Region Stockholm (ALF project), King Gustaf the V-80 years foundation, the Swedish Rheumatism Association and the Landspitali University Hospital Research Fund.