An Electron Microscope Study of the Development of a Mouse Hepatitis Virus in Tissue Culture Cells

• Published in: The Journal of cell biology 1965-01, Vol.24 (1), p.57-78
• Main Authors: David-Ferreira, J. F., Manaker, R. A.
• Format: Article
• Language: English
• Subjects: Cell membranes; Cells; Cultured cells; Cytoplasm; Cytoplasmic inclusions; Electrons; Hepatitis Viruses; L cells; Liver - cytology; Liver cells; Microscopy; Microscopy, Electron; Murine hepatitis virus; Old Medline; Tissue Culture Techniques; Virion; Virions; Virus Cultivation; Viruses
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.24.1.57

Samples taken at different intervals of time from suspension cultures of the NCTC 1469 line of mouse liver--derived (ML) cells infected with a mouse hepatitis virus have been studied with the electron microscope. The experiments revealed that the viruses are incorporated into the cells by viropexis within 1 hour after being added to the culture. An increasing number of particles are found later inside dense cytoplasmic corpuscles similar to lysosomes. In the cytoplasm of the cells from the samples taken 7 hours after inoculation, two organized structures generally associated and never seen in the controls are observed: one consists of dense material arranged in a reticular disposition (reticular inclusion); the other is formed by small tubules organized in a complex pattern (tubular body). No evidence has been found concerning their origin. Their significance is discussed. With the progression of the infection a system of membrane-bounded tubules and cisternae is differentiated in the cytoplasm of the ML cells. In the lumen of these tubules or cisternae, which are occupied by a dense material, numerous virus particles are observed. The virus particles which originate in association with the limiting membranes of tubules and cisternae are released into their lumen by a "budding" process. The virus particles are 75 mμ in diameter and possess a nucleoid constituted of dense particles or rods limiting an electron transparent core. The virus limiting membrane is sometimes covered by an outer layer of a dense material. In the cells from the samples taken 14 to 20 hours after inoculation, larger zones of the cell cytoplasm are occupied by inclusion bodies formed by channels or cisternae with their lumens containing numerous virus particles. In the samples taken 20 hours or more after the inoculation numerous cells show evident signs of degeneration.