Extended Data Fig. 5: Intact cell extrinsic determinants of B cell development and function.
(a) Levels of sCD40L, APRIL and BAFF in the plasma of CBL-LOH patients (n = 6) and healthy donors (n = 14). Line shows the mean of the displayed datapoints (one point per individual assessed). (b) Quantification of the production of sCD40L, APRIL and BAFF by PBMCs of CBL-LOH patients (n = 3) and healthy donors (n = 5). Line shows the mean of the displayed datapoints (one point per individual assessed). (c-f) Impact of sCD40L levels on B cell differentiation in vitro using CD34+ HSPCs. (c,d) B cell output at day 21 of co-culture of CD34+ HSPCs from two healthy donors. The MS5 co-culture was supplement with IL-7 (20 ng/mL) and the indicated doses of sCD40L. Mean of dots that represent technical replicates. (e,f) Quantification of B cell subsets based on CD10 and CD20 marker expression. Cells were treated as in (c,d). Mean ± s.d. (g) IL-21 production by sorted CD4+ naïve and memory T cells upon the indicated stimuli. Mean ± s.d. (h) Surface staining of CD38 expressed on CD19+ HSPC-derived B cell progenitors edited with the indicated sgRNAs. Mean ± s.d. of three biological replicates. (i) Surface staining of CD38 expressed on HEK293T, REH and BJAB cells as determined by flow cytometry. Mean ± s.d. of three biological replicates. (j) CBL Y371C KI BJAB (n = 9) cells are not more resistant than WT (n = 5) cells to BCR-induced apoptosis. Mean ± s.d. of clones over three independent experiments.
