Extended Data Fig. 4: Flow cytometry gating strategy. | Nature Immunology

Extended Data Fig. 4: Flow cytometry gating strategy.

From: Memory CD8+ T cell diversity and B cell responses correlate with protection against SARS-CoV-2 following mRNA vaccination

Extended Data Fig. 4: Flow cytometry gating strategy.The alternative text for this image may have been generated using AI.

a,Gating strategy ancestry: lymphocytes (FSC-A/SSC-A), FSC singlets (FSC-W/FSC-A), SSC singlets (SSC-W/SSC-A), Dump- live cells [Dead FVS450+ cells and CD14+ CD16+ CD19+ CD56+ (Dump+) cells excluded], CD4 vs CD8 (BV605-CD8/BV786-CD4). b, c, Differentiation markers on CD4+ (b) or CD8+ (c)T cells. CD4+ or CD8+ T cells, CD25 vs CD69 (BV-510-CD25/APC-R700-CD69). CD4+ or CD8+ T cells with high gMFI levels for both CD25 and CD69 were gated as double positive cells. CD45RA (APC-CD45RA/FSC-A) on gated CD4+ or CD8+ and on gated CD25+ CD69+ CD4+ or CD8+ T cells, CCR7 vs CD27 (BV711-CCR7/ APC-H7-CD27) on gated CD45RA− and on gated CD45RA+ cells. d, e, Flow cytometry analysis showing the percentage of CD4+ (d) and CD8+ (e) T cells in VNLRs (n = 25), VNHRs (n = 24), and VR subjects (n = 12) at month 6. f, g, Representative flow cytometry dot plots of CD25 vs CD69 cells on gated CD4+ (f) and CD8+ (g) T cells from VNLRs, VNHRs and VR subjects at T0, after 24h incubation with Medium (Unstimulated) or SWT. Data are represented with background subtraction from paired unstimulated controls. The same subjects were longitudinally analyzed. Box plots indicate median, interquartile range, and minimum/maximum. Each dot of box plots represents the average between three technical replicates of the same subject. All data are pooled from multiple experiments. Statistics were calculated using two-sided Wilcoxon rank-sum tests.

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