Poor Yield of Dendritic Cell Precursors from Untreated Pediatric Cancer

Abstract

We have studied the generation of dendritic cells (DC) in vitro from healthy children and children with newly diagnosed cancer. Peripheral blood derived adherent cells were harvested and cultured in the presence of granulocyte-macrophage colony-stimulating-factor (GM-CSF) and interleukin-4 (IL-4). Differentiated DC were characterized morphologically and analyzed by flow cytometry and allogenic mixed lymphocyte reaction (MLR). The numbers of adherent cells were two-fold higher in healthy children than in those with malignant tumors: 1.5 × 10⁵/ml of blood (mean) versus 0.7 × 10⁵/ml, respectively (p = 0.025). No significant differences were found in the cell survival or yield after the in vitro cultivation of adherent cells. Cytological examination of cultured cells showed that they were similar to DC in adults, being large, irregularly shaped, with several thin membrane protrusions, and bean-shaped nuclei. Differentiated DC from healthy controls expressed CD86 and HLA-DR, but did not express monocyte markers CD14 and CD64 (FcγRI). The phenotype of DC from cancer patients was otherwise similar, except that a substantial proportion (24–85%) continued to express CD64 (p = 0.001). DC derived both from cancer patients and controls were strong stimulators in allogeneic MLR. We conclude that functionally capable DC can be generated in vitro from blood-derived adherent cells in children, but children with untreated cancer yield lower numbers of DC than healthy children. The continued expression of CD64 on DC derived from cancer patients may indicate that adherent cells from cancer patients are more resistant to signals inducing differentiation into DC.