Fig. 4.
Removal CD19-CAR-T pressure restores endogenous expression of CD19 on transferred CD19-downregulated B cells A Experimental outline. Images generated with BioRender.com. B Gating strategy for sorting CD19-lo and CD19-int B220 + B cells isolated from lymph nodes and spleens from T-cell deficient animals 10 days after adoptive transfer of 4 × 106 CD19-CAR-T cells. Sample pooled from 3 CD19-CAR-T treated animals. C Sorted 2.2 × 105 CD19-lo or CD19-int B cells were adoptively transferred into congenically distinct wild-type hosts. Lymph nodes and spleens from adoptively transferred animals were harvested 60 days after transfer for flow cytometric analysis. Representative plots of endogenous B cell populations, transferred CD19-int B cells, and transferred CD19-lo B cells isolated from pooled lymph node and spleen are shown (n = 3 per transfer group, n = 6 for endogenous populations, from 1 independent experiment). D CD19 frequency (left panel) and gMFI (right panel) of B220 + cells isolated from pooled lymph node and spleen from animals adoptively transferred with sorted CD19 B cell populations (n = 3 per transfer group, n = 6 for endogenous populations, from 1 independent experiment). Statistical analysis was performed using one-way ANOVA