Morphological, Chemical, and Biological Characterization of Japanese Encephalitis Virus Virion and Its Hemagglutinin

Abstract

Three morphologically distinct structures, inner core, envelope, and surface projections, were observed in purified Japanese encephalitis virus virions by electron microscopy. The average diameter of each structure was 29.8 ± 2.5, 44.8 ± 3.2, and 53.1 ± 4.5 nm, respectively. Double staining with uranyl acetate and phosphotungstic acid preserved these structures well. Treatment of virions with proteolytic enzymes resulted in the loss of hemagglutinating activity, surface projections, and the major polypeptide band in polyacrylamide gel electrophoresis, which corresponds to glycoprotein, one of the three virion polypeptides. Surface projections were purified by cesium chloride density gradient centrifugation after treatment of virions with Nonidet P-40. The purified materials had a density of 1.256 g/cm 3 and were composed of only glycoprotein, as revealed by polyacrylamide gel electrophoresis. Purified surface projections carried hemagglutinating activity, as well as neutralizing antibody-blocking activity, and induced neutralizing antibody in mice.