Down-regulation of L-selectin Surface Expression by Various Leukocyte Isolation Procedures

Autor: C Bührer, D Stibenz
Rok vydání: 1994
Předmět:
Zdroj: Scandinavian Journal of Immunology. 39:59-63
ISSN: 1365-3083
0300-9475
DOI: 10.1111/j.1365-3083.1994.tb03340.x
Popis: L-selectin, a cell surface glycoprotein expressed on lymphocytes, granulocytes, and monocytes, has been implicated in lymphocyte homing and extravasation of phagocytic leukocytes into areas of inflammation. Considerable differences of L-selectin expression among various individuals has been reported, with clinical correlations to perinatal events, maturation, and circadian rhythm. In this study, L-selectin expression of various white blood cells was found to be differentially sensitive to ficoll-hypaque or percoll density gradient centrifugation. After density gradient centrifugation, a significant loss of median monocyte L-selectin expression was observed when compared to time and temperature-matched controls or results obtained by whole blood incubation with anti-L-selectin monoclonal antibodies followed by simultaneous leukocyte fixation and red cell lysis. Mock treatment itself was associated with a variable L-selectin loss of monocytes but not lymphocytes or granulocytes. Ficoll-hypaque or percoll density gradient centrifugation resulted in significant L-selectin down-regulation of lymphocytes while granulocytes separated from lymphocytes and monocytes by ficoll-hypaque or percoll retained full L-selectin surface reactivity. L-selectin downregulation was seen also after colloid sedimentation with hydroxy-ethyl starch. It is concluded that unseparated blood should be used for measuring L-selectin expression.
Databáze: OpenAIRE