Electroporation of dendritic cells with autologous total RNA from tumor material.

Dendritic cells (DC) are unique antigen-presenting cells that initiate and orchestrate adaptive immunity. Theoretically, cancer cells that express tumor-specific antigens can be destroyed by cytotoxic T cells. However, inherent antitumor responses are often not efficient, since tumor cells can mask their antigens and do not activate DC, an event required for the development of tumor antigen-specific cytotoxic T cell responses. Over a decade ago, the ex vivo preparation of autologous tumor antigen-loaded monocyte-derived DC vaccines as a novel potent anticancer strategy was launched. Phase I and II trials have been performed employing this strategy to treat several malignancies, such as B cell lymphoma, myeloma, melanoma, prostate, colon, ovarian, pancreatic, breast cancer, and renal cell carcinoma. So far, DC immunotherapy is well tolerated with little side or toxic effects. An issue of concern is the way DC are loaded with tumor antigens. An effective strategy is the loading of DC with tumor antigen through electroporation with tumor RNA. In this chapter, a comprehensive description of a protocol for loading of ex vivo-derived DC with total tumor RNA through electroporation is provided.