Nephrol Dial Transplant (1996) 11: 2453-2460
© 1996 European Renal Association-European Dialysis and Transplant Association
research-article
Activation of human neutrophils after contact with cellulose-based haemodialysis membranes: intracellular calcium signalling in single cells
1Institut für Immunologie Heidelberg 2Deutsches Krebsforschungszentrum Heidelberg, Germany 3University of Maryland at.Baltimore, Medical School, Dept. of Pathology Baltimore, MD, USA 4Gambro Co., Hechingen Germany 5Department if Internal Medicine Heidelberg, Germany 6Institut für Pharmazeutische Technologie and Biopharmazie Heidelburg, Germany
Correspondence and offprint requests to: Correspondence and offprint requests to: Gertrud M. Hänsch PhD, Instiut fü Immunologie der Universität Heidelberg, Im Neuenheimer Feld 305,69120 Heidelberg, Germany
PURPOSE OF STUDY.: In vitro contact of human leukocytes with cellulose-based dialysis membranes under complement-independent conditions results in activation of various leukocyte functions. To analyse signals involved in the mechanism of cell activation, we measured changes in cytosolic free calcium ([Ca2+]i) in individual human blood neutrophils (PMN) upon contact with flat sheet haemodialysis membranes.
RESULTS.: By confocal laser-scanning microscopy (CLSM), changes in [Ca2+]i were monitored in Fluo-3-labelled cells up to 10 min after contact with a regenerated cellulose (RC) membrane. Multiple [Ca2+]i transients were observed for cells in contact with RC; biostochastic analysis showed that up to 67% of the PMN responded with a high increase in [Ca2+]i the rest were low- or non-responding cells. After contact with the new synthetic polycarbonate-polyether (PC-PE) membrane only non-responding cells were seen, indicating reduced cellular contact activation. The increase in [Ca2+]i of cells on RC could be inhibited by 5 mM L-fucose. This monosaccharide was recently found to bepresent in cellulose-based polymers in picomolar concentrations.
CONCLUSIONS.: The data supports the hypothesis that dialysis-membrane-associated L-fucose residues participate in complement-independent leukocyte activation during haemodialysis therapy.
Keywords: biocompatibility; calcium imaging; dialysis membranes; L-fucose; haemodialysis; cell activation
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