In general, vaccine formulations can be tested either for their ability to elicit anti-tumor protection or for eradication of established tumor. Although, the latter is the more challenging task, both assays are feasible using 38c-13 and A20 lymphomas. However, use of the slower-growing lymphoma, A20, is recommended for immunotherapy experiments. Both types of in vivo experiments, tumor protection and therapy of established tumor, are described in this protocol. Moreover, the protocol starts with description of immunization strategies for tumor protection experiments using DNA ("a" steps), protein ("b" steps), and cytokine-transduced vaccines ("c" steps), followed by tumor challenge protocols, including a tumor challenge protocol for protection studies ("d" step; same as tumor challenge steps in Basic Protocol 1) and a detailed experiment for immunotherapy of established A20 lymphoma ("e" steps). All in vivo experiments should be repeated at least three times with 10 mice per experimental group (6- to 12-week-old syngeneic female mice). Ig-KLH, whole-cell, or chemokine-sFv fusion protein vaccines require immunization once or twice every two weeks, while DNA vaccines require three or more immunizations every two weeks. Sera (200 |l) are drawn from the retro-orbital spaces of five randomly selected mice per group before each boost and tested for the presence of anti-idiotypic antibodies by ELISA (Support Protocol 2), to monitor immunization, prior to tumor challenge.
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