Tumor cell specific immune response.

<p><b>a</b>) Splenocytes from control and MIP treated tumor bearing mice at 3 weeks after tumor implantation were in-vitro restimulated with UV-irradiated B16F10 melanoma cells for 48 hrs. Culture supernatants were then analyzed for the levels of IL-12, IFN-γ, TNF-α, IL-10, IL-1β and IL-6 by ELISA. Each bar represents the mean ± SD of the two independent experiments (n = 4); (□) unstimulated; (▪) restimulated; **<i>p</i><0.01. <b>b</b>) NK cell cytotoxicity. To check NK cell cytotoxicity, CFSE labeled Yac-1 target cells were co-cultured with splenocytes from control and MIP treated tumor bearing mice for 6 hrs. Target cell killing was analysed on the basis of PI uptake by flow cytometry. Cytotoxicity was found to be higher in the MIP treated group as compared to control. <b>c</b>) CTL response against B16 melanoma cells. CTLs were derived from splenocytes of control and MIP treated tumor bearing mice by culturing with UV-irradiated B16F10 cells and rIL-2 for 5 days. Cytotoxicity was determined by co-culturing CTLs with CFSE-labeled B16F10 target cells. % Cytotoxicity = 100×[No. of CFSE & PI double positive cells]÷[(No. of CFSE positive cells)+(No. of CFSE & PI double positive cells)].