Heat-shocked tumor cell lysate-pulsed dendritic cells induce effective anti-tumor immune response in vivo

Figure 1 Western blot detection of HSP70 expression of CT-26 cells heating at 42 °C. 1: No heating. 2: heat-shocked for 30 min. 3: heat-shocked for 1 h. 4: heat-shocked for 2 h.

Figure 2 The surface molecule expressions of mouse bone marrow derived DCs were measured by staining with PE anti-mouse CD11c (N418, hamster IgG), FITC anti-mouse I-A/I-E (m5/114. 15.2, ratIgG2b), FITC anti-mouse CD86 (B7.1, PO3, ratIgG2b) and analyzed by FACS. 1: DC culture system at d 7. 2: DC culture system at d 9. 3: DC culture system stimulating with LPS(1 μg/mL). 4: DC culture system pulsed with CT-26 lysates. 5: DC culture system pulsed with HSCT-26 lysates. The data are representative for 3 independent experiments.

Figure 3 The concentration of mIL-12 p70 in supernatant of different DC culture system were measured by ELISA assay. 1: DC culture system at d 7. 2: DC culture system at d 9. 3: DC culture system pulsed with CT-26 lysates.4: DC culture system pulsed with HSCT-26 lysates. 5: DC culture system stimulating with LPS (1μg/mL). The data were representative for three independent experiments.

Figure 4 Tumor-specific IFN-γ secreting Splenic T cells induced by different cell lysates pulsed DC. 1: positive control (no target, 1mg/L PHA stimulated). 2: negative control (no target). 3: single DC immunized. 4:CT-26 lysates pulsed DC immunized. 5: HSCT-26 lysates pulsed DC immunized. Results are representative of three experiments.

Figure 5 Bone marrow-derived DC pulsed with HSCT-26 lysate can generate tumor-specific CTLs in vivo. Splenic T cells obtained from immunized mice were evaluated for cytolytic activity against Mitomycin C treated CT-26 cells at the effector: target (E:T) ratios indicated. Values are the mean ± SE of triplicate wells.

Figure 6 Survival curves of mice immunized with different dendritic cells tumor vaccine.