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Down‐regulation of L‐selectin Surface Expression by Various Leukocyte Isolation Procedures
Author(s) -
STIBENZ D.,
BÜHRER C.
Publication year - 1994
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1994.tb03340.x
Subject(s) - isolation (microbiology) , l selectin , expression (computer science) , selectin , biology , chemistry , microbiology and biotechnology , immunology , computer science , inflammation , cell adhesion molecule , programming language
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‐hypaqus or percoll retained full L‐seleclin 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.