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Therefore, another control experiment was set up in which Vac-LMP2A-immunized mice were challenged with either Renca-LMP2A or Renca-VC tumors. The data show that mice challenged with Renca-VC MRIP not only had significantly higher tumor volumes, P?=?0.0009, (Fig.?(Fig.4D)4D) at all the time points tested, but the time to reach MATS was significantly reduced, P?=?0.006, (Fig.?(Fig.4E),4E), compared with mice challenged with Renca-LMP2A. The uncontrolled growth of Renca-VC tumors, compared with Renca-LMP2A tumors, in immunized mice confirmed the specific CTL targeting of LMP2A epitopes expressed by the Renca-LMP2A tumors. The data thus far show that, despite the presence of tumor-infiltrating LMP2A-specific CTL (generated by Vac-LMP2A immunization), the fact that there is slow but progressive growth of Renca-LMP2A tumors clearly suggests that there is only partial control of tumor growth. EtxB has been shown to colocalize with LMP2A in the lipid rafts of EBV-infected LCL where it enhances antigen presentation leading to increased CTL targeting of the LCL 14. We have also demonstrated enhanced targeting of LMP2A in vitro following treatment of Renca-LMP2A with EtxB (Fig.?(Fig.3D3D find more and ?andE).E). We next tested the ability of EtxB to enhance the in vivo antitumor response to Renca-LMP2A. Once again, groups of BALB/c mice were either immunized with Vac-LMP2A to generate an underlying LMP2A-specific CTL response or left unimmunized as controls. Eleven days later, these mice were challenged with either Renca-LMP2A in PBS (PBS controls) or Renca-LMP2A in PBS containing 50?��g of EtxB. The mice PD173074 cost were continued on either PBS or EtxB treatment twice a week for the first 10?days. As soon as the tumors became palpable (from day 10 after tumor challenge), EtxB or PBS was administered daily for 8?days, as outlined in Figure?Figure5A.5A. EtxB delivery was then stopped on day 18 and the rest of the animals culled; except for the immunized mice treated with EtxB, in which tumor measurements were taken for a further 10?days in the absence of EtxB treatment. Figure 5 Enhanced immune control of tumor growth by frequent administration of EtxB. (A) Schedule for immunization and EtxB (or PBS) for experiments shown in B�CF. (B and D) Unimmunized or (C, E, and F) Vac-LMP2A-immunized mice were injected with Renca-LMP2A ... Among the unimmunized mice, there was no difference in the time taken for tumor onset or in the rate of tumor progression; whether or not mice were treated with either EtxB or PBS (Fig.?(Fig.5B5B and ?andC).C). However, Vac-LMP2A-immunized mice that were treated with EtxB showed a 5-day delay in tumor onset which was accompanied by significantly lower tumor volumes (P?