A Couple Of Concepts To Simplify Erastin
On Days 11, 18, and 25, 106 DCs were injected into the tumor (intratumor). Two days after the last injection of DCs (on Day 27), 106 SCCVII cells were injected subcutaneously into the contralateral (left) thigh. http://www.selleckchem.com/products/3-deazaneplanocin-a-dznep.html Tumor volume was calculated by using the following formula: 0.5236 (long axis) �� (short axis).2, 14, 15 The volume of the primary tumor on the right thigh was measured every 3 or 4 days. The size of tumors was determined 30 days after the challenge, and tumors http://www.selleck.cn/products/pf-06463922.html with Versalyse (Beckmann Coulter, Fullerton, Calif), splenocytes (4 �� 106 cells/mL) from individual mice were cocultured separately and restimulated for 5 days with SCCVII cells (3 �� 105 cells/mL) that had been treated previously with mitomycin C (2 mg/mL). During restimulation, the cells were cultured in the presence of IL-2 (30 U/mL; PeproTec) for 3 days. The cells were harvested on Day 6 and were used as effector cells in a standard chromium 51 (51Cr) release assay, as described previously.15 Maximum release and spontaneous release were defined as the counts from samples incubated with 2% Triton X-100 or medium alone, respectively. Cytolytic activity was calculated using the following formula: percentage of specific 51Cr release = (experimental release ? spontaneous release) �� 100/(maximum release ? spontaneous release). An immunoglobulin fraction was collected from ascites of Balb/c nude mice that had been injected with a hybridoma line TIB-222 (anti-CD8). Antiasialo GM1 polyclonal http://www.selleckchem.com/products/erastin.html antibody was purchased from WAKO Chemicals (Osaka, Japan). Anti-CD8 monoclonal antibody (250 ��g per dose), antiasialo GM1 (100 ��g per dose), or phosphate-buffered saline was injected intraperitoneally for 1 week every second day into the treated mice, which had completely rejected the second challenge of SCCVII. Two days after the last injection of antibody (110 days after primary tumor inoculation), 106 SCCVII cells were inoculated into the abdomen as a third challenge. The Fisher exact test was used to evaluate the acceptance or rejection of the multiple tumor challenges. Multivariate analysis of the factors that affected inhibition of the second inoculation of SCCVII was performed to identify independent DC-related factors (iDC or rSeVDC, requirement of tumor lysate, and route of DC administration), and hazard ratios were assessed using a Cox proportional hazards model.