Tumor growth was apparently inhibited by Tpit/E vaccination. C. Series of CT scan selleck images showing growth of the subcutaneous tumor. (a-d): a control mouse; day 10 (a), 14 (b), 17 (c) and 21 (d) after tumor challenge, (e-h): a Tpit/E vaccinated mouse; day 10 (e), 21 (f), 24 (g) and 28 (h) after tumor challenge. Tumors were indicated by arrowheads. Arrows in panel g. and h. point to necrotic region in the tumor. D. Survival rate after tumor challenge; p < 0.05: Tpit/E vaccine vs. control.
Inhibition of lung metastasis by the Tpit/E vaccination B16/F10 cells were injected into the tail vein on the same schedule as the subcutaneous tumor model and development of lung metastasis was followed by CT scanning. At day 7 after tumor challenge, no metastasis was detected in all mice. At day STI571 ic50 14, metastases appeared in all of control mice and three out of eight Tpit/E
vaccination mice (Table 1). Metastasis appearance rate at this time point was significantly inhibited by Tpit/E cell vaccination. Series of CT images at day 14 and 21 post tumor challenge of representative mice of each group are shown in Fig. 3A. Survival period of the Tpit/E cell vaccination find more group was significantly longer than control (Fig. 3B). Table 1 Animals with lung metastases at day 14 post tumor challenge mice with metastasis mice without metastasis control 6 0 T-pit/E vaccine 3 5 Animals with lung metastases at day 14 post tumor challenge; p < 0.05: control vs. T-pit/E vaccine by the chi-square analysis. Figure 3 Tumor growth and survival rate in the lung metastasis model. A. Series of CT scan images showing development of the lung metastases, (a, b): a control mouse, (c, d): a Tpit/E vaccinated mouse; day 14 (a, c) and 21 (b, d) after tumor challenge. Anidulafungin (LY303366) Tumors were indicated by arrowheads.
No lung metastasis was observed in panel c. B. Survival rate after tumor challenge, p < 0.05: Tpit/E vaccine vs. control. Anti-endothelial cell specific antibody generated in a Tpit/E vaccinated mouse To make sure that specific antibodies to Tpit/E cells are generated in a mouse vaccinated with Tpit/E cells, we aimed to obtain Tpit/E specific antibody-secreting hybridoma clones. A surviving mouse in the Tpit/E cell vaccination group of the subcutaneous tumor model was sacrificed at day 45 after tumor challenge. Hybridomas of the spleen cells and SP-2 cells were prepared and the conditioned media were subjected to immunostaining to check the presence of IgG reactive to Tpit/E or B16/F10 cells. Some hybridoma clones were shown to secret antibodies reactive to Tpit/E but not to B16/F10 cells. Images of immunostaining with the medium of a representative clone along with phase contrast images are shown (Fig. 4). Edge of the colonies of Tpit/E cells was strongly stained, while B16/F10 cells were not stained. Figure 4 Antibody to Tpit/E cells generated in the vaccinated mouse.