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. 2021 Apr 12;76(10):1010–1019. doi: 10.1136/thoraxjnl-2020-216256

Figure 2.

Figure 2

BALF T cells comprise predominantly effector memory CD4 and CD8 T cells and CD8 Trm. PBMCs and BALFMCs isolated from patients with COVID-19 admitted to the ICU were measured using spectral flow cytometry (n=17). T cells were phenotyped (see online supplemental table 3 for all subsets) using CD27, CD28, CCR7 and CD45RA and CD95 in naïve (CD45RA+CD27+CD28+CD95−), stem cell-like memory (SCM; CD45RA+CD27+CD28+CD95+), effector memory-1 (EM1) (CD45RA−CD27+CD28+CCR7−), EM2 (CD45RA−CD27−CD28+CCR7+), EM3 (CD45RA−CD27−CD28+CCR7−), EM4 (CD45RA−CD27−CD28−CCR7−), effector memory Ra+ (TEMRA; CD45RA−CD27+CD28+CCR7−), central memory (CM; CD45RA−CD27+CD28+CCR7+), tissue-resident memory (Trm; CD103+CD28−; only for BALFMCs) and regulatory T cells (Treg; CD25+CD127−; only for CD4 T cells) (A, F). Activation (ie, HLA-DR+CD38+) is presented for different CD4 and CD8 T cell subsets (only for populations with >250 events) (B, F). Representation of PD-1 expression on different T cells subsets (C, G) in PBMC and BALFMC with concomitant quantification of total PD-1 expression (D, H,). Levels of IL-4 (I), IL17-a (J), granzyme B (K), IL-2 (L), IL-7 (M), IL-10 (N) and soluble PD-L1 (O) are presented in plasma and BALF. Box plots represent median±IQR. anti-N, anti-nucleacapsid; anti-RBD, anti-receptor binding domain of spike protein; BALF, bronchoalveolar lavage fluid; DN, double negative; ICU, intensive care unit; PBMC, peripheral blood mononuclear cells; PD-1, programmad death-1; PD-L1, programmed death-ligand 1. Statistical significance was tested with multiple testing correction using Kruskal-Wallis (A, B, E, F: control vs patient PBMC), Friedmans test (A, B, E, F: patient PBMC vs BALFMC), Wilcoxon signed-rank test (D, H, I–O: COVID-19 plasma vs BALF) or Mann-Whitney U test (I–O: healthy vs COVID-19). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.