TY - JOUR
T1 - Chemical agents and enzymes used for the extraction of gut lymphocytes influence flow cytometric detection of T cell surface markers
AU - van Damme, N.
AU - Baeten, D.
AU - de Vos, M.
AU - Demetter, P.
AU - Elewaut, D.
AU - Mielants, H.
AU - Verbruggen, G.
AU - Cuvelier, C.
AU - Veys, E. M.
AU - de Keyser, F.
PY - 2000
Y1 - 2000
N2 - 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
AB - 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
U2 - https://doi.org/10.1016/S0022-1759(99)00243-4
DO - https://doi.org/10.1016/S0022-1759(99)00243-4
M3 - Article
C2 - 10699577
SN - 0022-1759
VL - 236
SP - 27
EP - 35
JO - Journal of immunological methods
JF - Journal of immunological methods
IS - 1-2
ER -