Figure 1.

CyTOF panel and workflow analysis delineates four monocyte subsets in peripheral blood. (A) Biaxial plots showing the expression of markers on Ir⁺CD45⁺ PBMCs measured by mass cytometry. A representative HD is shown. An arcsinh scale (−5.0 to 10⁴) with a cofactor of 5 was used. (B) By mass cytometry analysis >100,000 Ir⁺CD45⁺ cells were defined on a biaxial plot before classification with the viSNE algorithm. MPS (>20,000 cells) was gated as remaining cells after the exclusion of B (CD19⁺), T (CD3⁺), and NK (CD3⁻CD16⁺CD45RA⁺) lymphocytes and doublets (see Supplemental Fig. 1). (C) Events in the MPS gate were then parsed with SPADE into 30 nodes using all markers for clustering, except CD19 and CD3. CD14⁺, CD16⁺, and Slan⁺ SPADE groups were observed to match classic (CD14⁺CD16⁻), intermediate (CD14⁺CD16⁺), nonclassic Slan^low (CD14^dimCD16⁺Slan^low), and nonclassic Slan⁺ (CD14^dimCD16⁺Slan^high) monocytes. A representative HD is shown. % represents the frequency among PBMCs. (D) On the 4 monocyte subsets previously described in (B), heat maps show the relative normalized transformed mean intensity for various markers tested by mass cytometry, for a representative HD.