Hemodialysis-induced degranulation of polymorphonuclear cells: no correlation between membrane markers and degranulation products.
OBJECTIVE
Degranulation of polymorphonuclear leukocytes (PMN) during hemodialysis (HD) is usually assessed by measuring degranulation products. However, this process might also be estimated by the assessment of cell surface markers. In this study, the relationship between the expression of PMN degranulation markers (CD63 and CD66b) and the release of degranulation products [myeloperoxidase (MPO) and lactoferrin (LF)] was investigated during clinical HD in order to evaluate cell surface markers as a useful index of PMN degranulation.


METHODS
The expression of CD63 and CD66b on PMN and the release of MPO and LF were investigated in 10 chronic HD patients, during both heparin (HDhep) and trisodium citrate anticoagulation (HDcit), in a randomized order. Samples were drawn from both the efferent and afferent lines of the dialyzer at 0, 7.5, and 180 min.


RESULTS
During HDhep at first passage, a major increase in MPO (from 158 +/- 32 to 448 +/- 177 microg/l, p = 0.001) and LF (from 134 +/- 52 to 260 +/- 120 microg/l, p = 0.01) was found across the dialyzer, whereas marked changes were not observed during HDcit. The expression of CD63 and CD66b increased across the dialyzer during both anticoagulation modalities, but was only significant in the case of HDhep (CD63: mean fluorescence intensity from 247 +/- 61 to 331 +/- 118, p < 0.01; CD66b: mean fluorescence intensity from 340 +/- 76 to 434 +/- 103, p = 0.01). During HDhep a correlation was noted between the degranulation products and markers of both azurophilic and specific granules (MPO and CD63: r = 0.35; p < 0.01; LF and CD66b: r = 0.39, p < 0.01). Significant differences in the expression of CD63 and CD66b between HDhep and HDcit were not observed. When analyzing the combined data for both HDhep and HDcit, no correlation was observed between degranulation products and markers.


CONCLUSIONS
Our data suggest that the measurements of cell surface markers may not be a reliable indicator of the degree of HD-induced PMN degranulation.
Grooteman
M P
MP
Medical Center Alkmaar, The Netherlands.
van Tellingen
A
A
van Houte
A J
AJ
Bos
J C
JC
Schoorl
M
M
van Limbeek
J
J
Nubé
M J
MJ
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000925:Anticoagulants / Q000627:therapeutic use; D015703:Antigens, CD / Q000378:metabolism; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D015550:Cell Degranulation; D002951:Citrates / Q000627:therapeutic use; D005260:Female; D058851:GPI-Linked Proteins; D006493:Heparin / Q000627:therapeutic use; D006801:Humans; D007781:Lactoferrin / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D008875:Middle Aged; D009504:Neutrophils / Q000276:immunology / Q000502:physiology*; D009195:Peroxidase / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D006435:Renal Dialysis / Q000009:adverse effects*; D000077559:Sodium Citrate; D060149:Tetraspanin 30
D000925:Anticoagulants; D015703:Antigens, CD; D000951:Antigens, Neoplasm; C554586:CD63 protein, human; C504120:CEACAM8 protein, human; D015815:Cell Adhesion Molecules; D002951:Citrates; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D000077559:Sodium Citrate; D006493:Heparin; D009195:Peroxidase; D007781:Lactoferrin