Fig 3. Aberrant BACH2 expression induces proliferation and suppresses effector fates in primary CD4 + T cells.
(a) To mimic aberrant BACH2 expression, CD3/CD28-activated CD4 + T cells were transduced with LV-BACH2 and control LV-GFP lentivirus and assessed longitudinally for proliferative behavior (flow-cytometry based) and transcriptomic changes. Created in BioRender. Brauckmann, L. (2026) https://BioRender.com/1vu1fc4. (b) Schematic of LV-BACH2 and LV-GFP expression cassettes. Created in BioRender. Brauckmann, L. (2026) https://BioRender.com/s96zj7i. (c) Cell tracer-based assay for proliferative behavior of LV-BACH2, LV-GFP and non-transduced cells. Cells were stained with CellTrace Far Red at d12 post transfection and followed on d14, d15 and d17. Statistical analysis was done using unpaired t-test, *p < 0.05, **p < 0.03. (d) Growth competition of LV-BACH2 and LV-GFP transduced CD4 + T cells (n = 3 donors) in co-culture. Shown are percentages of LV-BACH2 and LV-GFP cells at days (d) 7, 11 and 18 post initiation of co-culture. (e) Principal component analysis based on RNAseq datasets of SPB-dLNGFR-enriched LV-BACH2 and LV-GFP transduced CD4 + T cells at d4 (n = 3), d9 (n = 4) and d25 (n = 3) post transduction. (f) Number of differentially expressed genes (DEG; cutoffs set at |log2FC| > 1, p-adj < 0.05) between LV-BACH2 and LV-GFP transduced CD4 + T cells at d4, d9 and d25 post transduction. (g) Gene set enrichment analysis (GSEA) based on DEGs between LV-BACH2 and LV-GFP transduced CD4 + T cells at d9 post transduction delineates suppressed cellular pathways in LV_BACH2 cells. (h) STRING analysis based on DEG between LV-BACH2 and LV-GFP transduced CD4 + T cells at d25 post transduction.