LDL Receptor Activity in Human Leukocyte Subtypes: Regulation by Insulin

Objectives: LDL receptors of leukocytes play a key role in lipoprotein uptake, immunoregulation and the pathogenesis of atherosclerosis. Numerous studies with different methods of low reliability yielded conflicing results of its regulation in leukocyte subtypes. Design and Methods: LDL receptors of human leukocytes were measured with use of the monoclonal antibody C-7. Specific C-7 binding was detected by FACS analysis using phycoerythrin-anti-mouse-IgG. Parallel incubations with FITC-labelled anti-LEU 4 (CD 3), anti-LEU 12 (CD 19) and anti-MY 4 (CD 14) antibodies were used to distinguish C-7 binding of specific cell types (T-, B-lymphocytes and monocytes). Results: In contrast to monocytes, T and B-lymphocytes freshly isolated from healthy blood donors had no detectable binding capacity for C-7. After 24 and 48 h incubation of cells in a lipid-free medium, lymphocytes acquired some C-7 binding, albeit still much less than monocytes. Incubation with insulin for 24 h in a concentration of 0.5 μg/mL led to an increase in C-7 binding for monocytes (up to 180%). Saturation experiments with the ligand suggests an increase in the number of receptors. In contrast the same insulin concentration inhibited C-7 binding of B- and T-lymphocytes by 35%. Conclusions: FACS analysis using monoclonal antibodies seems to be a feasible method for the investigation of lipid metabolism in leukocytes. The LDL receptor expression and its regulation by insulin differs in circulating monocytes and lymphocytes.