Photosensitizer targeting in photodynamic therapy. II. Conjugates of haematoporphyrin with serum lipoproteins.
Conjugates between haematoporphyrin (HP) and human low-density lipoprotein (LDL), human high-density lipoprotein (HDL) and bovine HDL have been prepared, purified and characterized. HP-LDL is aggregated possibly via interparticle apoB protein cross-linking. HP-HDL human and bovine conjugates show different degrees of intraparticle apoA polypeptide cross-linking. Receptor-mediated endocytosis of HP-LDL by NIH 3T3 cells is inferred from the increased uptake observed when LDL receptors are upregulated. HP-LDL uptake into HT29 cells faces competition from unlabelled LDL, albeit at rather high doses. HP-HDL uptake is also inhibited by LDL, suggesting that both lipoprotein conjugates may have cell-surface binding sites in addition to the specific LDL (apoB) receptor. J774.2 macrophages avidly accumulate HP-LDL, retaining most of the fluorescence and some of the protein while degrading the remainder. Oxidized LDL species compete in these processes, with the major effect on protein degradation. Chloroquine has little effect on the fluorescence uptake but inhibits protein degradation (and hence enhances protein accumulation). HP-HDL is also avidly taken up by J774.2 cells, but in the case of the bovine material with a sigmoidal concentration dependence. This is consistent with prior aggregation before the particles can be endocytosed. P388.D1 cells, which appear to be less activated than the J774.2 line, take up less fluorescence and retain and degrade less protein, but still to higher extents than observed for non-phagocytic cells. We conclude that photosensitizer-lipoprotein conjugates can be taken up in large amounts by cells possessing scavenger receptors and/or phagocytic activity, and that this may be a means of targeting photodynamic therapy.
Hamblin
M R
MR
Department of Surgery, University of Dundee, Ninewells Hospital, UK.
Newman
E L
EL
D016475:3T3 Cells; D000818:Animals; D001667:Binding, Competitive; D001692:Biological Transport; D002417:Cattle; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004337:Drug Carriers; D004705:Endocytosis; D006415:Hematoporphyrins / Q000378:metabolism* / Q000633:toxicity; D006801:Humans; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism* / Q000633:toxicity; D008077:Lipoproteins, LDL / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism* / Q000633:toxicity; D051379:Mice; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000378:metabolism* / Q000633:toxicity; D011973:Receptors, LDL / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured
D004337:Drug Carriers; D006415:Hematoporphyrins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D017319:Photosensitizing Agents; D011973:Receptors, LDL