[Determination of polymorphonuclear neutrophil adhesion receptors. Effect of pre-analytic factors].
Polymorphonuclear neutrophil (PMN) adherence receptors expression varies with leukocyte activation state. Their quantification need accurate and inter-laboratories reproducible methods, without artefactual activation.


OBJECTIVE
The aim of this study was to study the influence of cell preparation on PMN adherence receptors expression.


METHODS
It was proposed to quantify, using immunolabeling standard (QIFIKIT(R), Dako), surface expression of the main adherence receptors (L-selectin and B(2) integrins), from different preparations of PMN: total blood collected with EDTA, isolated PMN by density gradient Polymorphprep(TM) 1,113 (Nycomed Pharma) and formaldehyde fixed PMN.


RESULTS
A decrease of all receptors was noted after isolation and fixation of PMN, in comparison with whole blood PMN analysis. These results differed from data previously reported since, in these studies, activated phenotypes (increased of B(2) integrins) were observed after isolation and fixation methods.


CONCLUSIONS
The present study provides strong evidence that pre-analytical conditions are sources of biological variations and thus extreme care must be taken in the interpretation of results. It underlines the interest of consensual practices for these pre-analytic and analytic parameters in order to compare results in multicenter and longitudinal studies.
Latger-Cannard
V
V
Regnault
V
V
Dumas
D
D
Nguyen
P
P
Lecompte
T
T
Stoltz
J
J
D018821:CD18 Antigens / Q000097:blood*; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods; D006801:Humans; D019041:L-Selectin / Q000097:blood*; D018375:Neutrophil Activation; D009504:Neutrophils / Q000166:cytology / Q000502:physiology*; D016707:Tissue Fixation / Q000379:methods
D018821:CD18 Antigens; D019041:L-Selectin