Detection of tissue factor-positive extracellular vesicles using the ExoView R100 system

Background: Tissue factor (TF) is essential for hemostasis. TF-expressing extracellular vesicles (TF⁺ EVs) are released in pathological conditions, such as trauma and cancer, and are linked to thrombosis. Detection of TF⁺ EV antigenically in plasma is challenging due to their low concentration but may be of clinical utility.

Objectives: We hypthesised that ExoView can allow for direct measurement of TF⁺ EV in plasma, antigenically.

Methods: We utilized the anti-TF monoclonal antibody 5G9 to capture TF EV onto specialized ExoView chips. This was combined with fluorescent TF⁺ EV detection using anti-TF monoclonal antibody IIID8-AF647. We measured tumor cell-derived (BxPC-3) TF⁺ EV and TF⁺ EVs from plasma derived from whole blood with or without lipopolysaccharide (LPS) stimulation. We used this system to analyze TF⁺ EVs in 2 relevant clinical cohorts: trauma and ovarian cancer. We compared ExoView results with an EV TF activity assay.

Results: BxPC-3-derived TF⁺ EVs were identified with ExoView using 5G9 capture with IIID8-AF647 detection. 5G9 capture with IIID8-AF647 detection was significantly higher in LPS+ samples than in LPS samples and correlated with EV TF activity (R² = 0.28). Trauma patient samples had higher levels of EV TF activity than healthy controls, but activity did not correlate with TF measurements made by ExoView (R² = 0.15). Samples from patients with ovarian cancer have higher levels of EV TF activity than those from healthy controls, but activity did not correlate with TF measurement by ExoView (R² = 0.0063).

Conclusion: TF⁺ EV measurement is possible in plasma, but the threshold and potential clinical applicability of ExoView R100, in this context, remain to be established.