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. 2020 Dec 16;218(3):e20200547. doi: 10.1084/jem.20200547

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© 2020 Lu et al.

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Figure 4. — Mice lacking follicular regulatory CD4⁺ T cells exhibit a distinct BCR repertoire following influenza virus infection. (A) Principal component analysis visualization of V_H gene usage frequency for total unique Igh sequences isolated from the mLN-derived repertoire. (B) Heatmaps showing V_H gene usage frequencies for the top 15 V_H genes based on maximal principal component 2 score from principal component analysis of total Igh sequences. These are shown with values normalized for each row based on Z-score. (C) Individual V_H gene frequencies relative to the total number of unique variant VDJ sequences are plotted for the top six V_H genes. Significance was assessed using a two-tailed Wilcoxon test with a significance threshold of *, P < 0.1; **, P, < 0.05. Horizontal bars show the average frequency of V_H gene usage for samples of a specific genotype. (D) Scatterplots for V_H genes based on their principal component 2 loading score compared with their usage frequency among the published set of V_H sequences from Kuraoka et al. (2016) associated with HA specificity (compared with non-HA specifics; left) or with PA specificity (compared with non-PA specific sequences; right) at day 16 after immunization. Correlations were computed as Pearson correlation coefficients, and a P value was assigned to the significance of the correlation with a significance threshold of P < 0.05. For the BCR repertoire analysis, data are from one experiment with four mice per group performed 15 d following PR8 influenza virus infection. Cor., Pearson correlation coefficient.