LIPID UTILIZATION BY HUMAN-LYMPHOCYTES IS CORRELATED WITH HIGH-DENSITY-LIPOPROTEIN BINDING-SITE ACTIVITY

The nature and physiological importance of high-density lipoprotein (HDL) binding sites on unstimulated (resting) and mitogen-activated (blast) human peripheral blood lymphocytes were investigated. Specific HDL binding on resting and blast T-lymphocytes was saturable at 50-mu-g of I-125-HDL/ml and of high affinity, with K(d) values of 8.1 x 10(-8) M and 6.5 x 10(-8) M, respectively, and B(max) values of 79 ng and 180 ng/mg of cell protein respectively at 4-degrees-C. Binding of HDL double-labelled with fluorescent dioctadecylindocarbocyanine (Dil) and isotope (I-125) as well as of single fluorescence- or isotope-labelled HDL was inhibited competitively by HDL apoproteins. Studies of the cholesterol flux between the cells and HDL showed that HDL, low-density lipoprotein (LDL) or BSA at a concentration of 100-mu-g/ml in the tissue culture medium did not result in a significant difference in exogenous [H-3]cholesterol efflux from the cell membrane at 37-degrees-C. Proliferating T-blasts incorporated more cholesterol from HDL or LDL than did resting lymphocytes. When the cells were pulsed with I-125-HDL and chased in fresh lipid-free medium, up to 80 % of the radioactivity released was not precipitable with trichloroacetic acid. This percentage decreased in a competitive manner when unlabelled HDL was present in the chase incubation medium. Finally, cultivation of lymphocytes with conditioned medium from macrophages increased Dil-HDL binding/uptake, while it was decreased by mevinolin-induced inhibition of hydroxymethylglutaryl-coA reductase. In conclusion, human lymphocytes possess a HDL binding site (receptor) responsible for lipid binding/uptake and concomitant internalization and degradation of apoproteins from HDL, but not for reverse cell membrane cholesterol transport. The activity of the binding site is up-regulated during cell proliferation and down-regulated during cell growth suppression.