Role of complement and platelet-activating factor in the stimulation of phagocytosis and reactive oxygen species production during haemodialysis.
BACKGROUND
Neutrophil phagocytic functions have been studied extensively in haemodialysis (HD) patients; however, results are contradictory and the mechanisms that modulate phagocytosis and oxidative burst during dialysis are not completely understood.


METHODS
The present study investigated neutrophil functions in a selected population of patients before and during clinical dialysis with cuprophane, and polyacrylonitrile (AN69) membranes. We measured phagocytosis of Escherichia coli and intracellular hydrogen peroxide (H2O2) production by flow cytometry in whole blood.


RESULTS
Before dialysis, neutrophils from HD patients showed normal phagocytic capability and H2O2 formation. Phagocytosis of FITC-E. coli was significantly stimulated in cuprophane but not AN69-treated patients. Spontaneous and stimulated H2O2 production was enhanced with both cuprophane and AN69 membranes. We then investigated in vitro the role of complement and platelet-activating factor (PAF) in the activation of neutrophils. Incubation of whole blood with C5a increased phagocytosis but not H2O2 production. On the contrary, the addition of synthetic PAF showed a markedly stimulated H2O2 production without increase in phagocytosis. Moreover, during dialysis with formaldehyde-reused cuprophane, complement activation was abolished and phagocytosis was no longer enhanced, while the stimulation of H2O2 production persisted. In addition, we also excluded a particular role of the membrane itself in the activation of neutrophils.


CONCLUSIONS
We demonstrated that in a selected population of HD patients, neutrophils exhibit normal phagocytic capability and normal intracellular H2O2 production. During dialysis, the stimulation of phagocytosis observed with cuprophane is complement dependent, whereas the enhanced H2O2 production observed with both cuprophane and AN69 membranes might be related to PAF production.
Gastaldello
K
K
Department of Nephrology, Dialysis and Transplantation, Hôpital Erasme, Université Libre de Bruxelles, Belgium. kgastald@ulb.ac.be
Husson
C
C
Wens
R
R
Vanherweghem
J L
JL
Tielemans
C
C
D003167:Complement Activation / Q000502:physiology; D003165:Complement System Proteins / Q000502:physiology*; D004731:Endotoxins / Q000097:blood; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008567:Membranes, Artificial; D009504:Neutrophils / Q000378:metabolism / Q000502:physiology; D010587:Phagocytosis / Q000187:drug effects / Q000502:physiology*; D010972:Platelet Activating Factor / Q000494:pharmacology / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012016:Reference Values; D006435:Renal Dialysis; D013997:Time Factors
D004731:Endotoxins; D008567:Membranes, Artificial; D010972:Platelet Activating Factor; D017382:Reactive Oxygen Species; D003165:Complement System Proteins; D006861:Hydrogen Peroxide