Human monocytes and keratinocytes in culture ingest hydroxyethylstarch

Severe itching for unknown reasons has been reported after administration of hydroxyethylstarch (HES) in haemodilution therapy of humans. In skin sections from patients who had received HES for otological reasons HES deposition was found by immunohistochemical methods in keratinocytes, epidermal dendritic cells, endothelial cells and macrophages. The aim of the present investigation was to study the interaction of cultured human monocytes and keratinocytes, normal (HNK) and malignant (KB, A431), with FITC-conjugated HES (200/0.5). Peripheral blood monocytes from healthy donors were isolated by Ficoll-Hypaque gradient centrifugation and adherence to plastic surfaces. Cells were incubated with HES-FITC (1 mg/ml) for 10, 20, 30, 60 and 120 min at 37 degrees C, washed several times to minimize cell-associated HES-FITC. The uptake of HES-FITC (200/0.5) was detected by fluorescence microscopy and flow cytometry. Monocytes and keratinocytes (HNK, KB, A431) acquired significant amounts of HES-FITC after 120 min. Endocytosis was blunted by the addition of unconjugated HES in ten-fold concentration. At 4 degrees C, a further inhibition was achieved, indicating an uptake of HES-FITC by monocytes and keratinocytes in vitro. These results indicate that a connection between the storage of HES by keratinocytes and monocytes and the observed itching in patients might be possible.