Fig 2: scRNA-seq analysis of splenocytes from mice treated with 3A5 BiTE(A) Immune cells of the spleen from SFTSV-infected mice following treatment with control BiTE or 3A5 BiTE were isolated for scRNA-seq analysis (n = 3 per group). The UMAP plot displays all the immune cells from the two groups, colored by eight major cell types.(B) Proportion of the eight major cell types in the control BiTE group and 3A5 BiTE group.(C) Violin plots show cytotoxicity (left) and exhaustion (right) module score for CD4+ T (n = 936 and n = 532) and CD8+ T (n = 1997 and n = 1174) cells in the 3A5 BiTE-treated group and the control group, respectively. The horizontal lines display the 25th, 50th (median), and 75th percentiles. The cytotoxicity module score was calculated based on the expression levels of Gzma, Gzmb, Gzmk, Gzmc, Gzmm, Gzmf, Ifng, Nkg7, Prf1, Ccl4, and Ccl5. The exhaustion module score was calculated based on the expression levels of Pdcd1, Lag3, Ctla4, Havcr2, and Tnfrsf9. A two-tailed t test was performed to analyze the differences between two groups. ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns: no significance.(D) The UMAP plot displays T cells colored by 10 T cell subsets.(E) The bubble plot shows expression of well-documented marker genes in T cell subsets. Dot size represents gene expression percentage per cell type, with marker gene intensity displayed.(F) Comparison of the proportions of T cell subsets between control BiTE group and 3A5 BiTE group. Data were presented as mean ± SD Mann-Whitney test was used for comparisons of values among two groups. ∗p < 0.05; ∗∗p < 0.01; ns: no significance.(G) Heatmap of GSEA pathway enrichment scores of individual T cell clusters between 3A5 BiTE and control BiTE groups.(H) A circle plot showing differential interaction strength among CD4+ T cells, CD8+ T cells, B cells, and plasma cells across two groups. Red edges indicate stronger interactions in the 3A5 BiTE group in comparison with control BiTE group, while blue edges indicate stronger interactions in the control BiTE group compared with 3A5 BiTE group.(I) Spleen sections collected from 3A5 BiTE- and control BiTE-treated mice at 6 days post-infection were subjected to co-immunofluorescence staining using anti-CD4 and anti-CD19 antibodies. Scale bars, 100 μm.See also Figures S13 and S14.