Ann Lab Med.  2024 Sep;44(5):392-400. 10.3343/alm.2023.0395.

Platelet and Monocyte Microvesicles as Potential Biomarkers of COVID-19 Severity: A Cross-Sectional Analysis

• Nunki N
• Hernaningsih Y
• Wardhani P
• Herawati A
• Yusoff NM
• Moses EJ
• Semedi BP

• KMID: 2559145
• DOI: http://doi.org/10.3343/alm.2023.0395

Abstract

Background
Coronavirus disease (COVID-19) induces inflammation, coagulopathy following platelet and monocyte activation, and fibrinolysis, resulting in elevated D-dimer levels. Activated platelets and monocytes produce microvesicles (MVs). We analyzed the differences in platelet and monocyte MV counts in mild, moderate, and severe COVID-19, as well as their correlation with D-dimer levels.
Methods
In this cross-sectional study, blood specimens were collected from 90 COVID-19 patients and analyzed for D-dimers using SYSMEX CS-2500. Platelet MVs (PMVs; PMVCD42b⁺ and PMVCD41a⁺ ), monocyte MVs (MMVs; MMVCD14⁺ ), and phosphatidylserinebinding annexin V (PS, AnnV⁺ ) were analyzed using a BD FACSCalibur instrument.
Results
PMV and MMV counts were significantly increased in COVID-19 patients. AnnV⁺ PMVCD42b⁺ and AnnV⁺ PMVCD41a⁺ cell counts were higher in patients with severe COVID-19 than in those with moderate clinical symptoms. The median (range) of AnnV⁺ PMVCD42b⁺ (MV/µL) in mild, moderate, and severe COVID-19 was 1,118.3 (328.1–1,910.5), 937.4 (311.4–2,909.5), and 1,298.8 (458.2–9,703.5), respectively (P = 0.009). The median (range) for AnnV⁺ PMVCD41a⁺ (MV/µL) in mild, moderate, and severe disease was 885.5 (346.3–1,682.7), 663.5 (233.8–2,081.5), and 1,146.3 (333.3–10,296.6), respectively (P = 0.007). D-dimer levels (ng/mL) weak correlated with AnnV⁺ PMVCD41a⁺ (P = 0.047, r = 0.258).
Conclusions
PMV PMVCD42b⁺ and PMVCD41a⁺ counts were significantly increased in patients with severe clinical symptoms, and PMVCD41a⁺ counts correlated with D-dimer levels. Therefore, MV counts can be used as a potential biomarker of COVID-19 severity.

Keyword

Annexin V; CD14; CD41a; CD42b; COVID-19; D-dimer; Microvesicles

Figure

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  Fig. 1 Flow-cytometric detection of MVs. (A) The MV gate region was separated, defined, and calculated using the SSC-height (log) scale as the threshold to eliminate the 0.16-µm beads and use the 0.2- and 0.5-µm bead clouds to set the MV gate. The gate used for capturing the beads for counting is depicted as region 1 (R1). (B) MVs were calibrated using Megamix-Plus SSC beads in a Trucount tube, using fluorescence (FL1)/FITC as the threshold. (C) FSC and SSC of isolated MVs stained with fluorescence (FL), fluorescein isothiocyanate (FITC)-conjugated CD42b, and Trucount calibrator beads (R1). (D) Analysis of a specimen from a COVID-19 patient using a Trucount tube to obtain the R4 region. R4 comprised Trucount gating beads for counting. (E) MV-negative control, analyzed from healthy controls without staining with fluorescent-labeled antibodies (unstained). (F) The MV population of healthy controls with annexin V-positive staining was analyzed for PMVs (AnnV+ PMVCD42b+, AnnV+ PMVCD41a+) and MMVs (AnnV+ MMVCD14+). (G) The MV population of mild COVID-19 patients with annexin V-positive staining was analyzed for PMVs and MMVs. (H) The MV population of moderate COVID-19 patients with annexin V-positive staining was analyzed for PMVs and MMVs. (I) The MV population of severe COVID-19 patients with annexin V-positive staining was analyzed for PMVs and MMVs. The detected MVs were conjugated with fluorescent FITC for CD42b, PerCP-Cy5.5 for CD41a, APC for CD14, and PE for annexin V. Abbreviations: MVs, microvesicles; PMVs, platelet microvesicles; MMVs, monocyte microvesicles; Qty, quantity; FSC, forward scatter; SSC, side scatter; FITC, fluorescein isothiocyanate; PerCP-Cy5.5, peridinin chlorophyll protein-cyanine5.5; APC, allophycocyanin; PE, phycoerythrin.

Reference

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