Fig. 2: Phenotyping of peripheral blood monocytes in VEXAS syndrome.

A Proportions (frequencies) of classical (CD14⁺, CD16⁻), intermediate (CD14⁺, CD16⁺), and nonclassical (CD14^lo, CD16⁺) monocytes among blood monocytes from VEXAS patients, VEXAS-like patients, MDS and healthy controls, were analyzed (CD14⁺, CD16⁺, *P  =  0.00379; CD14^lo, CD16⁺, **P  =  0.0084). Each dot represents a single patient. B Non-supervised Uniform Manifold Approximation and Projection (UMAP) of blood monocytes. Cells are automatically separated into spatially distinct subsets according to the combination of markers that they express. C UMAP of blood monocytes stained with 13 markers and measured with mass cytometry. D UMAP colored according to cell density across patients’ groups. Red indicates the highest density of cells. E Volcan plots of differentially represented monocyte subsets showing clusters increased or decreased in VEXAS patients compared to healthy controls, and VEXAS patients compared to VEXAS-like. F Heatmap representation of all monocyte clusters, ordered by hierarchical clustering and expression of the cell surface markers. G Proportion (frequencies) of CXCR3 + CXCR5+ expressing monocytes. Each dot represents a single patient. Gating strategy for the analysis of expression of CXCR3 and CXCR5 on blood monocytes, Illustrative dot plots are shown and enumeration of percentages of CXCR3 + CXCR5+ cells (**P  =  0.0012 and **P  =  0.0048). H Pie chart showing the proportion of HLA-DR^lo dysfunctional monocytes, exhausted monocytes expressing chemokine receptors, HLA-DR^hi functional monocytes, and other clusters not significantly different between groups. Numbers on the pie charts indicate the median proportion of each monocytes subsets in each group. P values were determined by the two-sided Kruskal-Wallis test, followed by Dunn’s post test for multiple group comparisons. *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001.