Pozzi D, Lisi A, De Ros I, Ferroni L, Giuliani A, Ravagnan G, Grimaldi S
Istituto di Medicine Sperimentale C.N.R., Roma, Italy.
Photochem Photobiol. 1993 Mar;57(3):426-30. doi: 10.1111/j.1751-1097.1993.tb02313.x.
Human erythrocytes were separated into five fractions representing different age groups. In each group phospholipid inside-outside translocation was determined by quantitation of the amino phospholipids phosphatidylserine and phosphatidylethanolamine and their lyso-derivatives by thin layer chromatography. To assess the role of transbilayer phospholipid distribution in the recognition and fusion of vesicular stomatitis virus (VSV) and human aged erythrocytes, we monitored the fusion kinetics using the octadecylrhodamine dequenching assay. Fusion of VSV with each single group of red blood cells (RBC) was not detectable with the youngest cells (F1 group) but increased with RBC aging (F2-F5 groups). The same increase in fusion was observed with microvesicles generated from RBC in which aging was mimicked by incubating the cells with Ca2+ in the presence of the Ca2+ ionophore A23187. Conversion of the aminophospholipids to the trinitrophenyl derivative by reaction with trinitrobenzensulfonate completely inhibits fusion on ghosts in which aging was artificially induced by translocation of aminophospholipids in the outer leaflet (symmetric ghosts). These results indicate that RBC become susceptible to VSV fusion during aging and in all pathology related to the aging process.
人红细胞被分离成代表不同年龄组的五个部分。在每组中，通过薄层层析法定量测定氨基磷脂磷脂酰丝氨酸和磷脂酰乙醇胺及其溶血衍生物，来确定磷脂的内外转运。为了评估跨膜磷脂分布在水疱性口炎病毒（VSV）与衰老人红细胞的识别和融合中的作用，我们使用十八烷基罗丹明去淬灭测定法监测融合动力学。VSV与最年轻的细胞（F1组）融合未被检测到，但随着红细胞衰老（F2 - F5组）融合增加。在用钙离子载体A23187存在下用Ca2 +孵育细胞模拟衰老的红细胞产生的微泡中也观察到相同的融合增加。通过与三硝基苯磺酸盐反应将氨基磷脂转化为三硝基苯基衍生物，完全抑制了通过在外层小叶中转运氨基磷脂人工诱导衰老的血影（对称血影）的融合。这些结果表明，红细胞在衰老过程中以及在与衰老过程相关的所有病理状态下变得易于VSV融合。
