Fig. 4. CLL cell engraftment in NSG mice conditioned with HS5-CD40L-IL4 or HS5-CD40L-IL4-IL21 cells.

A A scheme of the transplantation approach. The scaffold was incubated with HS5-CD40L-IL4 or HS5-CD40L-IL4-IL21 cells in vitro and transplanted s.c. on the left flank of NSG mice (6–8 weeks old). After 7 days, each mouse was injected with HS5-CD40L-IL4 ± IL21 cells i.p.. At 14 days, CLL cells (purity >98% CD5⁺CD19⁺ cells) were mixed with HS5 cells and injected s.c. and i.p.. Control mice were injected with purified CLL cells only (day 14, not shown). B Human CD20 staining of tumor B cells in mice co-transplanted with CLL and HS5-CD40L-IL4 cells. Magnification 200×. C Spleen weight of control mice and mice co-transplanted with HS5-CD40L-IL4 or HS5-CD40L-IL4-IL21 cells with or without detectable tumor growth. D Representative images of the spleen of control mice (left) and mice with engrafted tumors (right). E CD20 staining of tumor B cells in mice co-transplanted with HS5-CD40L-IL4 or HS5-CD40L-IL4-IL21 cells (control stands for mice transplanted with CLL cells only, no HS5 cells co-transplanted). F Spleen (i) and tumor (ii) weight and relationship to the mice’s time to death. Mice that died spontaneously are not included due to missing spleen weight values. G Time from CLL cell transplantation into NSG mice conditioned with HS5-CD40L-IL4 (n = 10) or HS5-CD40L-IL4-IL21 (n = 5) cells to euthanasia due to extensive tumor growth. Mice with no tumor growth are not included in the graph. H FISH detection of 13q14 deletion in spleen sections from M20 and M29 mice with engrafted human B cells. Red dot, 13q14 probe; blue dot, 13q34 probe for control region; green dot, CEP12 control probe.