Enhanced T‐cell activation by immature dendritic cells loaded with HSP70‐expressing heat‐killed melanoma cells

Please cite this paper as: Enhanced T‐cell activation by immature dendritic cells loaded with HSP70‐expressing heat‐killed melanoma cells. Experimental Dermatology 2010; 19: 108–116. Abstract: Vaccination protocols that utilize dendritic cells (DCs) to elicit therapeutic immunity against tumors are the subject of intense research. Given that the capacity of DCs to cross‐present antigens is physiologically low, there is considerable interest to develop strategies that enhance that pathway. In order to best exploit the enhanced cross‐presentation of antigens bound to heat shock protein 70 (HSP70), we analysed melanoma cell preparations for their HSP70 expression. Western blotting revealed strong upregulation of HSP70 after heat‐killing in contrast to UV‐B irradiation. When the uptake of heat‐killed necrotic cells by DCs at various levels of maturation was assessed, 61 ± 7% of immature DCs (iDCs) internalized fluorescence‐labelled necrotic material. Apoptotic material from UV‐B‐irradiated cells was internalized by only 48 ± 5% of iDCs. Maturation‐inducing cytokines did not affect the uptake when added simultaneously with the tumor cell preparations. Loading DCs with heat‐necrotic or apoptotic melanoma cells slightly reduced CD83 expression while leaving CD208 (DC‐LAMP) expression unchanged. As determined by IFN‐γ‐detecting enzyme‐linked‐immunospot assays, iDCs loaded with heat‐killed melanoma cells activated autologous T cells most effectively when used without any further maturation, whereas DCs loaded with apoptotic material required maturation. In conclusion, HSP70‐expressing melanoma cells could be generated by heat‐killing. Loading iDCs with heat‐killed melanoma cells resulted in a superior priming of autologous T cells in vitro.