title: Lipid utilization by human lymphocytes is correlated with high-density-lipoprotein binding site activity.
abstract: 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 micrograms of 125I-HDL/ml and of high affinity, with Kd values of 8.1 x 10(-8) M and 6.5 x 10(-8) M, respectively, and Bmax. 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 (125I) 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 micrograms/ml in the tissue culture medium did not result in a significant difference in exogenous [3H]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 125I-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.
authors.affiliation: Institute for Biomedical Aging Research, Austrian Academy of Sciences, Innsbruck.
authors.forename: Q
authors.initials: Q
authors.lastname: Xu
authors.forename: G
authors.initials: G
authors.lastname: Jürgens
authors.forename: L A
authors.initials: LA
authors.lastname: Huber
authors.forename: G
authors.initials: G
authors.lastname: Böck
authors.forename: H
authors.initials: H
authors.lastname: Wolf
authors.forename: G
authors.initials: G
authors.lastname: Wick
mesh_terms: D000328:Adult; D001665:Binding Sites; D001692:Biological Transport; D002232:Carbocyanines / Q000737:chemistry; D002455:Cell Division; D002784:Cholesterol / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D007457:Iodine Radioisotopes; D050356:Lipid Metabolism; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000378:metabolism; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism*; D008297:Male; D013696:Temperature
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; D007457:Iodine Radioisotopes; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; C024286:3,3'-dioctadecylindocarbocyanine; D002784:Cholesterol