Comparative binding of 125I-and 99mTc-d-labeled native and glycated low-density lipoprotein to human microvascular endothelial cells-potential for atherosclerosis imaging?
Native (n), glycated (g), and glycoxidated (go) low-density lipoproteins (LDL) were labeled with 125I or 99mTc, and the labeling efficiency and binding were assessed for potential use of these LDL compounds in imaging analysis of atherosclerotic lesions (PPAR-gamma receptors) by determining the number of specific receptors for nLDL, gLDL or goLDL on human microvascular endothelial cells as well as the KDs using either 125I-or 99mTc-labeled LDLs. The specific activity of labeled gLDL and goLDL was much higher (for goLDL 20 times higher) than that of nLDL. Gel filtration of labeled LDLs revealed, however, that 99mTc-g/goLDL is significantly degraded by the labeling reaction. No fragmentation was observed for 99mTc-nLDL and all the 125I-labeled LDL forms. Binding studies using both 125I-and 99mTc-nLDL indicated a weak binding affinity (KD 10- 7mol/L) to human microvascular endothelial cells. The binding affinity of 125I-g/goLDL to these cells was significantly higher (KD 10- 9mol/L) and could be increased further by preactivation of the endothelial cells using TNFalpha. Incubation with 99mTc-goLDL, however, did not result in specific binding of the ligand, possibly as a consequence of the fragmentation of the lipoprotein during the labeling. Scatchard transformation of the binding data with 99mTc-gLDL revealed the presence of only a few binding sites. This was in contrast to the results obtained with 125I-labeled gLDL, which revealed a much higher membrane density of scavenger receptors for this ligand. We conclude that for in vitro binding studies as well as for potential in vivo imaging, only 125I-labeled goLDL should be used, whereas nLDL may be applied as 125I-or 99mTc-labeled ligand.
Sobal
Grazyna
G
Department of Nuclear Medicine, Medical University of Vienna, Vienna, Austria. Grazyna.Sobal@meduniwien.ac.at
Menzel
Ernst Johannes
EJ
Sinzinger
Helmut
H
D000328:Adult; D050197:Atherosclerosis / Q000175:diagnosis*; D003952:Diagnostic Imaging; D042783:Endothelial Cells / Q000473:pathology*; D005260:Female; D006031:Glycosylation; D006801:Humans; D007457:Iodine Radioisotopes; D007553:Isotope Labeling / Q000379:methods; D008077:Lipoproteins, LDL; D008297:Male; D008833:Microcirculation / Q000473:pathology; D008875:Middle Aged; D011485:Protein Binding; D011973:Receptors, LDL / Q000032:analysis*; D013667:Technetium
D007457:Iodine Radioisotopes; D008077:Lipoproteins, LDL; D011973:Receptors, LDL; D013667:Technetium