An improved method for detection of low density lipoprotein receptor defects in human T lymphocytes.
Familial hypercholesterolemia (FH) results from an inherited functional defect of the low density lipoprotein (LDL) receptor and is complicated by premature atherosclerosis. FH diagnosis is obtained by sophisticated techniques or is suggested by clinical criteria. We have developed a technique based on flow cytometry for the measurement of DiI-labeled LDL uptake in human peripheral blood T lymphocytes left for 2 days in a lipoprotein-deficient culture medium. Flow cytometry allowed us to discriminate the uptake of DiI-LDL by T lymphocytes subpopulation from the uptake by the whole mononuclear population using a T cell-specific anti-CD3 antibody. The method appeared to be highly specific for the receptor-mediated pathway of LDL uptake as DiI-LDL uptake was inhibited in the presence of a 10-fold excess of unlabeled LDL and by EDTA. A good relationship was found between the uptake of DiI-LDL and 125I-labeled LDL degradation. The test was applied in three groups of patients: patients with normal cholesterol levels, patients with heterozygous FH, and patients with high cholesterol levels but without clinical criteria of FH. The mean fluorescence intensities were 23.1 +/- 8.9, 6.3 +/- 1.7, and 17.1 +/- 3.5 (mean +/- standard deviation), respectively. The ability to measure the fluorescence in T lymphocytes improved the discrimination between FH and non-FH subjects when compared with values obtained from the whole mononuclear cell population. These results suggest that our method could be useful for LDL receptor defects screening.
Verhoeye
F R
FR
Groupe d'Etude du Métabolisme Tumoral, A.S.B.L., Hôpital de Jolimont, Haine-Saint-Paul, Belgium.
Descamps
O
O
Husson
B
B
Hondekijn
J C
JC
Ronveaux-Dupal
M F
MF
Lontie
J F
JF
Heller
F R
FR
D000293:Adolescent; D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000097:blood* / Q000235:genetics; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood; D008875:Middle Aged; D011973:Receptors, LDL / Q000235:genetics*; D012016:Reference Values; D013601:T-Lymphocytes / Q000378:metabolism*
D008077:Lipoproteins, LDL; D011973:Receptors, LDL