Prevention of gastrointestinal tumors based on adenomatous polyposis coli gene mutation by dendritic cell vaccine
J. Clin. Invest. Toshio Iinuma, et al. 113:1307 doi:10.1172/JCI17323 [Go to this article.]

Figure 4
Reactivity of serum with tumor T cells. (A-D) Reactivity of serum from mouse treated with DC/T + IL-12 against the cells derived from various carcinoma cell lines. Tumor T cells and other irrelevant tumor cells (2 ∞ 105) were incubated with 50 ∝l of the serum at 4_C for 30 minutes, washed extensively, and then incubated with 2 ∝l of FITC-conjugated rat anti-mouse Ig antibody. Fluorescence histograms shown by the cells were obtained with FACS: (A) tumor T; (B) MC38 colon carcinoma; (C) B16 melanoma; (D) Hepa1-6 hepatocellular carcinoma. Thick line, negative control; dotted line, serum from untreated mouse; thin line, serum from mouse treated with DC/T + IL-12. (E) Decrease of serum reactivity by incubation with tumor T cells. Serum from a mouse treated with DC/T + IL-12 was diluted 100-fold with PBS, incubated with tumor T cells (2 ∞ 105) at 4_C for 1 hour and centrifuged. Tumor T cells were incubated with 50 ∝l of the supernatant (dashed line), 100-fold diluted serum (dotted line), or untreated mouse serum (thick line) at 4_C for 30 minutes, washed extensively, incubated with 2 ∝l of FITC-conjugated rat anti-mouse Ig antibody, and submitted for flow-cytometric analysis using FACS. Three independent experiments were performed with similar results. A typical experiment is shown. (F) Serum dilution and reactivity with tumor T cells. Tumor T cells (2 ∞ 105) were incubated with 50 ∝l of the diluted serum at 4_C for 30 minutes, washed extensively, and then incubated with 2 ∝l of FITC-conjugated rat anti-mouse Ig antibody. Fluorescence histograms shown by the cells were obtained using FACS, and the median fluorescence intensity was measured. All determinations were carried out in triplicate. Squares and triangles, sera from untreated mice; diamonds and circles, sera from mice treated with DC/T + IL-12.