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Chemical agents and enzymes used for the extraction of gut lymphocytes influence flow cytometric detection of T cell surface markers
Nancy Van Damme ,
Dominique Baeten ,
Martine De Vos ,
Pieter Demetter ,
Dirk Elewaut ,
Herman Mielants ,
Gust Verbruggen ,
Claude Cuvelier ,
Eric M. Veys ,
Filip De Keyser
March 2000
March 2000
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Abstract
Chemical agents (DTT, EDTA) and enzymes (collagenase, DNAse) are often used for the
generation of a single cell suspension from tissue samples. In this study, flow cytometry
was used to examine the effect of chemical agents and enzymes on the expression of
cell membrane markers of T lymphocytes from tonsils and peripheral blood. Expression
of CD4, CD8, CD25, CD38, L-selectin, CD44, alphaEbeta7 and alpha4beta7 were studied.
Incubation of lymphocytes with DTT and EDTA resulted in a decrease of CD38, alphaEbeta7
and alpha4beta7 expression. Incubation with collagenase A and DNAse resulted in a
decrease of CD25, L-selectin, alphaEbeta7 and alpha4beta7. The results of this study
indicate that a careful interpretation is necessary for phenotypic descriptions of
lymphocyte populations obtained by enzymatic isolation techniques.