Figure 5.

ITGB4-targeted immunotherapies elicited ITGB4-specific host immune responses against CSCs as well as non-CSCs. A, B: Splenetic T cells harvested from mice treated with ITGB4-DC vaccine (A) or mITGB4 BiAb-armed TDLN T cells (B) mediated significant cytotoxicity against 4T1 cells. Data were replicated in a second experiment for both ITGB4-DC vaccine and mITGB4 BiAb-armed TDLN T cell adoptive transfer experiments. C-F: ITGB4-targeted immunotherapy conferred significant host anti-ITGB4 humoral immunity in both 4T1 and SCC7 models. Immune plasma was collected from treated mice and IgG was quantified using ELISA. C: Plasma harvested from treated mice-bearing 4T1 tumor as indicated bound to 4T1 cells. D: Plasma IgG induced by ITGB4-DC vaccine bound to unsorted, ALDH^high, and ALDH^low SCC7 cells to different extend. E: Cytotoxic effects of immune plasma via CDC in 4T1 model. Plasma harvested from animals-bearing 4T1 tumor subjected to treatment was incubated with ALDH^high 4T1 or ALDH^low 4T1 cells for 1 hour followed by culturing in the presence of rabbit complement for another 1 hour. Lower percentage of viable cells at the end of incubation indicates more cell lysis. F: Similar CDC assay as in E was tested in the SCC7 model. Data were repeated twice.