Fig. 1: METTL1-associated translation is enhanced in B cells following vaccination.
a–g Single-cell RNA sequencing (scRNA-seq) analysis of PBMCs from donors before (d 0) and after (d 7) influenza vaccination (GSE211560). a t-SNE displaying cell clusters. b Bar graph displaying cell subset percentages before and after vaccination. c The top 20 terms upregulated in B cells compared with other clusters (adjusted p value via “clusterProfiler” GO analysis). d tRNA modification score (GO: 0006400) among cell clusters (4023 B cells, 29435 CD4 T cells, 7354 CD8 T cells, 4546 Monocytes, 6402 NK cells, 4613 NKT cells). The upper whisker is 75th percentile plus 1.5* interquartile range (IQR), and the lower is 25th percentile minus 1.5*IQR. The upper and lower bounds of box are 75th and 25th percentile, respectively, and the center is median. e Dot plot displaying differentially expressed genes (DEG) associated with common tRNA-modified enzymes. f Dot plot displaying Mettl1, Mettl8, Trmt1 and Nsun2 expression in B cells. g The top 10 GO-BP terms upregulated via GO analysis using upregulated DEGs on d 7 compared to d 0 B cells. h Experimental scheme of influenza vaccination. i METTL1 and WDR4 expression in B cells measured by western blotting. j, k The frequency of CD27hiCD38hi antibody-secreting cells (ASC) in PBMCs from donors receiving influenza vaccination (d 0 and d 28) measured by flow cytometry (n = 12). l H1N1, H3N2, and Victoria antibody titers in the sera of donors receiving influenza vaccination (d 0 and d 28), and n = 12. m–o scRNA-seq analysis of PBMCs from donors on the first day and 7 d after SARS-CoV-2 vaccination (GSE201534). m t-SNE displaying cell clusters from PBMCs. n Bar graph displaying cell subset percentages. o The top 5 terms upregulated in B cells compared with other clusters via GO analysis. p Experimental scheme of healthy donors receiving SARS-CoV-2 vaccination. q METTL1 and WDR4 expression in B cells measured by western blotting. r Representative flow cytometry plots of spike-specific B cells from the PBMCs of donors that received SARS-CoV-2 vaccination (d 0 and d 28). s, t Representative ELISpot and summary data showing spike- and receptor-binding domain (RBD)-specific IgG-secreting B cells from the PBMCs of donors who received SARS-CoV-2 vaccination (d 0 and d 28, n = 9). Biological independent samples for k, l, t. The data are presented as the mean ± SEM. A two-tailed unpaired Student’s t test in k, t, and Kruskal–Wallis test in l, d. Source data are provided as a Source Data file. h and p were created with BioRender.com.
