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Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation
Published on: July 8, 2011
Immune Monitoring Using mRNA-Transfected Dendritic Cells
Troels Holz Borch1,2, Inge Marie Svane3,4, Özcan Met3,4
1Department of Haematology, Center for Cancer Immune Therapy, Copenhagen University Hospital Herlev, Herlev Ringvej 75, 2730, Herlev, Denmark. troels.holz.borch@regionh.dk.
Abstract:
Dendritic cells are known to be the most potent antigen presenting cell in the immune system and are used as cellular adjuvants in therapeutic anticancer vaccines using various tumor-associated antigens or their derivatives. One way of loading antigen into the dendritic cells is by mRNA electroporation, ensuring presentation of antigen through major histocompatibility complex I and potentially activating T cells, enabling them to kill the tumor cells. Despite extensive research in the field, only one dendritic cell-based vaccine has been approved. There is therefore a great need to elucidate and understand the immunological impact of dendritic cell vaccination in order to improve clinical benefit. In this chapter, we describe a method for performing immune monitoring using peripheral blood mononuclear cells and autologous dendritic cells transfected with tumor-associated antigen-encoding mRNA.

