Differences in binding of platelet factor 4 to vascular endothelium in vivo and endothelial cells in vitro.
The binding of fluorescein-labelled recombinant human platelet factor 4 (rhPF4) to the vasculature of the hamster cheek pouch in vivo was compared with that to cultured endothelial cells (EC) from human umbilical veins (HUVEC) and arteries (HUAEC) and from human aorta (HAEC). In vivo data: systemically injected rhPF4 rapidly disappeared from plasma in a biphasic pattern (t1/2 = 2 and 41 min). High intensity non-uniform binding of rhPF4 occurred at short specific sites along both arterioles and venules. The length of the intense sites was 76 +/- 46 microns and their frequency was 10 +/- 4 per cm2 cheek pouch. Heparin was injected at 4 and 9 min, but not 30 min, post-rhPF4 displaced most of the high intensity labelling indicating internalization with time. Neither pretreatment with more than 50-fold excess of unlabelled rhPF4 nor histamine- or LTB4-induced vascular macromolecular leakage changed the frequency of short intense sites. In vitro data: uniform time-dependent intense binding of rhPF4 occurred in a similar fashion in subconfluent HUVEC, HUAEC and HAEC. All cell types showed nuclear staining, demonstrating internalization. When heparin was given to EC prior to rhPF4, binding was delayed in time but not blocked. In conclusion, rhPF4 does not bind uniformly with high intensity along pre- and post-capillary vessels of the hamster cheek pouch in vivo as predicted by the rhPF4-labelling of subconfluent (migrating/proliferating) human EC in vitro. The short infrequent sites of intense rhPF4-labeling in vivo may represent regions of endothelial cell migration/proliferation similar to subconfluent EC in culture.
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D001011:Aorta / Q000166:cytology; D001835:Body Weight; D002478:Cells, Cultured / Q000502:physiology; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D006493:Heparin / Q000494:pharmacology; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D007975:Leukotriene B4 / Q000494:pharmacology; D008297:Male; D008647:Mesocricetus; D018715:Microscopy, Video; D010978:Platelet Factor 4 / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding / Q000502:physiology; D014469:Umbilical Arteries / Q000166:cytology; D014471:Umbilical Veins / Q000166:cytology
D000925:Anticoagulants; D007975:Leukotriene B4; D010978:Platelet Factor 4; D006632:Histamine; D006493:Heparin; D016650:Fluorescein-5-isothiocyanate