Improved detection of familial hypercholesterolemia by determining low density lipoprotein receptor expression in mitogen-induced proliferating lymphocytes.

摘要: In view of the presence some 190 mutations in low density lipoprotein receptor (LDL-R) gene and a lack simple detection methods, we have developed an improved assay system for detecting familial hypercholesterolemia (FH) using mitogen-induced proliferating lymphocytes. Freshly isolated mononuclear cells were cultured 3 days RPMI 1640 supplemented with 10% human lipoprotein-deficient serum (LPDS) 1% phytohemagglutinin (PHA). LDL-R expression was measured by flow cytometry monoclonal anti-LDL-R antibody or DiI-LDL. Mitogenic responses monitored cell size (FSC), interleukin-2 (IL2-R) expression, stimulation index (SI). The PHA-stimulated lymphocytes significantly higher than monocytes without PHA (15.2- 3.6-fold, respectively). gradation highly correlated to FSC, IL2-R SI (r > 0.9 each case). However, no difference existed between 30 clinically diagnosed FH 42 normolipemic control subjects. lower group (45.2 +/- 15.3% versus 100 14.1%; unpaired t test, P < 0.0001) indicated genetic defects. Normocholesterolemic first degree relatives non-FH hypercholesterolemic subjects demonstrated normal as did controls. carries efficiency 97% both sensitivity specificity 98.5%. Measurement phytohemagglutinin- serum-stimulated offers method sensitivity.