Generation of Leukaemia-Derived Dendritic Cells (DC leu ) to Improve Anti-Leukaemic Activity in AML: Selection of the Most Efficient Response Modifier Combinations.

Dendritic cells (DC) and leukaemia derived DC (DC leu ) are potent stimulators of anti-leukaemic activity in acute myeloid leukaemia (AML) and can be generated from mononuclear cells in vitro following standard DC/DC leu -generating protocols. With respect to future clinical applications though, DC/DC leu -generating protocols specifically designed for application in a whole-blood-(WB)-environment must be established. Therefore, we developed ten new DC/DC leu -generating protocols (kits; Kit-A/-C/-D/-E/-F/-G/-H/-I/-K/-M) for the generation of DC/DC leu from leukaemic WB, containing calcium-ionophore, granulocyte-macrophage-colony-stimulating-factor (GM-CSF), tumour-necrosis-factor-alpha, prostaglandin-E 1 (PGE 1 ), prostaglandin-E 2 (PGE 2 ) and/or picibanil (OK-432). All protocols were evaluated regarding their performance in generating DC/DC leu using refined classification and/or ranking systems; DC/DC leu were evaluated regarding their performance in stimulating anti-leukaemic activity using a cytotoxicity fluorolysis assay. Overall, we found the new kits capable to generate (mature) DC/DC leu from leukaemic WB. Through refined classification and ranking systems, we were able to select Kit-I (GM-CSF + OK-432), -K (GM-CSF + PGE 2 ) and -M (GM-CSF + PGE 1 ) as the most efficient kits in generating (mature) DC/DC leu , which are further competent to stimulate immunoreactive cells to show an improved anti-leukaemic cytotoxicity as well. This great performance of Kit-I, -K and -M in mediating DC/DC leu -based anti-leukaemic immunity in a WB-environment in vitro constitutes an important and directive step for translating DC/DC leu -based immunotherapy of AML into clinical application.