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USE OF OCTADECYLRHODAMINE FLUORESCENCE DEQUENCHING TO STUDY VESICULAR STOMATITIS VIRUS FUSION WITH HUMAN AGED RED BLOOD CELLS *
Author(s) -
Pozzi Deleana,
Lisi Antonella,
Ros Ivan De,
Ferroni Lucietta,
Giuliani Anna,
Ravagnan Glanpietro,
Grimaldr Settimio
Publication year - 1993
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1993.tb02313.x
Subject(s) - vesicular stomatitis virus , fluorescence , fusion , virology , chemistry , virus , biology , physics , optics , linguistics , philosophy
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 (Fl 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 Ca 2+ in the presence of the Ca 2+ 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.