Expression of low-density lipoprotein receptors in peripheral blood and tonsil B lymphocytes.
B lymphocytes, purified from peripheral leucocytes from young normolipaemic humans, expressed and internalized low-density lipoprotein receptors (LDLR). The expression was assessed by a monoclonal anti-LDLR. The internalization of LDL was assessed by LDL labelled with 125I (125I-LDL) and 1,1'-dioctadecyl-3,3,3',3' tetramethyl-indocarboxycyanine perchlorate (LDL-DiI). The expression of LDLR, assessed by anti-LDLR, was: 38 +/- 8% (n = 5) for fresh purified cells, 60 +/- 10% (n = 12) for non-stimulated cells, 79 +/- 5% (n = 10) for IL-2 (100 U/ml)-stimulated cells and 95 +/- 5% (n = 8) for pokeweed mitogen (PWM) (1:200 dilution)-stimulated cells. The optimal concentrations of agonist were 100 U/ml of IL-2, and 1:200 dilution of PWM. IL-2 and PWM increased the internalization of LDL-DiI by 1.5-fold. The internalization of LDL-DiI was maximal at 60 microg of protein/ml (48 +/- 8%). Scatchard analysis revealed a Kd of 3.2 +/- 0.22 x 10(-8) M and 2180 +/- 190 binding sites in non-stimulated cells, a Kd of 7.73 +/- 0.36 x 10(-9) M and 12,500 +/- 430 binding sites for IL-2 (100 U/ml)-stimulated cells, and a Kd of 7.2 +/- 0.43 x 10(-9) M and 13,250 +/- 450 binding sites for PWM (1:200 dilution)-stimulated cells. Lineweaver-Burk analysis of LDL binding (LDL-DiI) revealed that the apparent Kd for non-stimulated cells was 1.3 +/- 0.11 x 10(-8) M, and 9.2 +/- 0.2 x 10(-9) M and 7.5 +/- 0.25 x 10(-9) M for IL-2- and PWM-stimulated cells, respectively. B lymphocytes from tonsils also showed a high expression of LDLR assessed with anti-LDLR (70 +/- 6%). The high expression of LDLR and the avid internalization of LDL suggest that LDL may be important for B cell physiological responses.
Institute of Immunology, Faculty of Medicine, Central University of Venezuela, Caracas.
J B
JB
De Sanctis
I
I
Blanca
H
H
Rivera
N E
NE
Bianco
D000328:Adult; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002469:Cell Separation; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000378:metabolism*; D014066:Palatine Tonsil / Q000166:cytology / Q000276:immunology*; D011043:Pokeweed Mitogens / Q000494:pharmacology; D011973:Receptors, LDL / Q000032:analysis*
D007376:Interleukin-2; D008070:Lipopolysaccharides; D008077:Lipoproteins, LDL; D011043:Pokeweed Mitogens; D011973:Receptors, LDL