title: Low density lipoprotein receptor expression and function in human polymorphonuclear leucocytes.
abstract: Low density lipoprotein receptors (LDLR), capable of internalizing LDL, are expressed in polymorphonuclear neutrophils (PMN). The expression was assessed using anti-LDLR antibody by flow cytometry. The internalization of LDL was assessed by: (i) quantification of the uptake of labelled LDL with 1,1'-dioctadecyl-3,3,3',3' tetramethyl-indocarboxycyanine perchlorate (DiI) by flow cytometry; and (ii) the binding of LDL-125I. In fresh purified cells, Lineweaver Burk analysis of LDL binding (LDL-DiI) revealed that the calculated Kd (internalized LDL) for PMN (15.0 x 10(-9) M) is lower than the Kd for monocytes (1.1 x 10(-7) M) and the Kd for lymphocytes (3.2 x 10(-7) M). Scatchard analysis (LDL-125I) revealed 25,000 binding sites and a Kd of 9.6 x 10(-9) M for PMN. The interaction of LDL with its receptor caused a two-fold fast (peak at 1 min) and transient increase in the oxidative burst, measured by the formation of 2',7' dicholoflurescein (DCF) by flow cytometry. This effect was not observed in monocytes or lymphocytes, and it was blocked by anti-LDLR antibody. The stimulation of LDL was optimal at 10 microg of protein/ml. LDL was able to suppress DCF formation induced by phorbol myristate acetate (PMA) and PMA was unable to further stimulate LDL-treated cells, suggesting protein kinase-C (PKC) involvement in LDL effects. Using a PKC assay, LDL was shown to induce a two-fold increase in PKC translocation to the membrane. Thus, LDL increases PMN oxidative burst through a PKC-dependent pathway.
authors.affiliation: Instituto de Immunología, Facultad de Medecina, Universidad Central deVenezuela, Caracas.
authors.forename: L L
authors.initials: LL
authors.lastname: Lara
authors.forename: H
authors.initials: H
authors.lastname: Rivera
authors.forename: C
authors.initials: C
authors.lastname: Perez-P
authors.forename: I
authors.initials: I
authors.lastname: Blanca
authors.forename: N E
authors.initials: NE
authors.lastname: Bianco
authors.forename: J B
authors.initials: JB
authors.lastname: De Sanctis
mesh_terms: D006801:Humans; D009504:Neutrophils / Q000378:metabolism*; D011973:Receptors, LDL / Q000096:biosynthesis* / Q000502:physiology*
chemical_list: D011973:Receptors, LDL