Comparison of low density lipoprotein uptake by different human lymphocyte subsets: a new method using double-fluorescence staining

A method to determine low density lipoprotein (LDL) uptake of distinct lymphocyte subpopulations was developed using fluorescent LDL and subsequent staining of lymphocyte subsets with biotinylated monoclonal antibodies plus streptavidin-CyChrome. LDL uptake was detected on a single cell level and semiquantified by FACS analysis. This method allows comparison of defined lymphocyte subsets from different individuals and excludes the falsifying influence of individual differences in subset distribution, which may occur in studies on total peripheral blood lymphocytes (PBL). Investigation of total PBL and lymphocyte subsets of 20 healthy volunteers (8 male, 12 female) showed the following. i) Different lymphocyte subsets exhibited highly significant differences in LDL uptake, with NK cells (CD16) showing a higher uptake than T (CD3) and B cells (CD19); CD8-positive cells exhibited higher values than CD4-positive cells. ii) These differences are due to specific, LDL-receptor (LDL-R)-mediated LDL uptake. iii) Inter-individual differences in LDL uptake are reflected on all lymphocyte subsets.