Mononuclear leucocyte function tests in the assessment of the biocompatibility of peritoneal dialysis fluids.
BACKGROUND
Previous studies showed that the currently used dextrose based peritoneal dialysis fluids impair several leucocyte functions.


OBJECTIVE
To determine which in vitro mononuclear leucocyte (monocyte) function tests most clearly reflect the biocompatibility of peritoneal dialysis fluid.


METHODS
Monocytes were tested for phagocytic capacity, bactericidal activity, Fc and C3 receptor expression, and chemiluminescence response, and by analysis of the release of interleukin 8 (IL-8) and tumour necrosis factor alpha (TNF alpha) in the presence of test fluids. Cytokine release was studied in an alternative dynamic in vitro peritoneal dialysis model in which monocytes were exposed to test fluid that was continuously equilibrated with an interstitial fluid-like medium through a microporous membrane. The chemiluminescence response by stressed monocytes was also tested after an 18 h recovery period. All tests were performed during or after exposure to different degrees of glycerol induced osmotic stress and after exposure to a 1% milk-whey derived, polypeptide enriched test fluid. Cells incubated in 0.1% gel Hanks buffer (GH) served as control.


RESULTS
Osmotic stress induced impairment of leucocyte function was found by the chemiluminescence assay (mean (SEM): 179 (20)% v 138 (23)% after 30 minutes in 0.5% and 1.5% glycerol, respectively) and by the analysis of IL-8 released by monocytes (44 (9) ng in 0.7% glycerol v 40 (7) ng in 2.0% glycerol). Only the chemiluminescence assay showed a protective effect of polypeptides on leucocyte function (after > or = 60 minutes). If monocytes were allowed to recover in culture medium after exposure to test fluids, the changes in chemiluminescence response appeared to be reversible after a 30 minute exposure, but became more pronounced after 60 and 120 minutes. The phagocytosis and bacterial killing assays were less sensitive. The observations carried out with the phagocytosis assay did not correspond with the Fc or C3 receptor density data.


CONCLUSIONS
The release of IL-8 by peripheral blood monocytes in a two compartment model and their chemiluminescence response are appropriate assays for the assessment of changes in leucocyte function in response to different peritoneal dialysis fluids.
Brulez
H F
HF
Department of Nephrology, Academisch Ziekenhuis Vrije Universiteit Amsterdam, The Netherlands. h.brulez@wxs.nl
ter Wee
P M
PM
Snijders
S V
SV
Donker
A J
AJ
Verbrugh
H A
HA
D015314:Dialysis Solutions / Q000494:pharmacology*; D006801:Humans; D016209:Interleukin-8 / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000502:physiology; D008163:Luminescent Measurements; D010587:Phagocytosis; D017463:Receptors, Complement 3b / Q000378:metabolism; D011961:Receptors, Fc / Q000097:blood; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D015314:Dialysis Solutions; D016209:Interleukin-8; D017463:Receptors, Complement 3b; D011961:Receptors, Fc; D014409:Tumor Necrosis Factor-alpha