title: Rotavirus is released from the apical surface of cultured human intestinal cells through nonconventional vesicular transport that bypasses the Golgi apparatus.
abstract: Rotaviruses are nonenveloped viruses that infect enterocytes of the small intestine and cause severe infantile gastroenteritis. It was previously thought that rotavirus exits cells by lysis, but this behavior does not match the local pathogenesis of the virus. In this study, we have investigated the release of the simian rotavirus strain (RRV) from the polarized intestinal Caco-2 cells. We found that RRV is released almost exclusively from the apical pole of Caco-2 cells before any cells lyse. Using confocal laser scanning microscopy and drugs that inhibit vesicular transport, we studied the RRV transport route from the endoplasmic reticulum (ER) to the apical side of intestinal cells. We demonstrated that RRV exits from the ER through a carbonyl cyanide m-chlorophenylhydrazone-sensitive vesicular transport. RRV staining was never found within the Golgi apparatus or lysosomes, suggesting that the RRV intracellular pathway does not involve these organelles. This finding was confirmed by treatment with monensin or NH4Cl, which do not affect release of RRV. Electron microscopic analysis revealed RRV containing small smooth vesicles in the apical area and free virions outside the cell in the brush border, consistent with a vesicular vectorial transport of virus. These results may provide, for the first time, a cellular explanation of the pathogenesis of rotavirus.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, Pathogénie Cellulaire et Moléculaire des Microorganismes Entérovirulents, Faculté de Pharmacie, Université Paris XI, Chatenay-Malabry, France.
authors.forename: N
authors.initials: N
authors.lastname: Jourdan
authors.forename: M
authors.initials: M
authors.lastname: Maurice
authors.forename: D
authors.initials: D
authors.lastname: Delautier
authors.forename: A M
authors.initials: AM
authors.lastname: Quero
authors.forename: A L
authors.initials: AL
authors.lastname: Servin
authors.forename: G
authors.initials: G
authors.lastname: Trugnan
mesh_terms: D001692:Biological Transport; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002462:Cell Membrane / Q000821:virology; D016764:Cell Polarity; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000821:virology; D019084:Fluorescent Antibody Technique, Indirect; D006056:Golgi Apparatus / Q000821:virology; D006801:Humans; D007413:Intestinal Mucosa / Q000821:virology*; D008247:Lysosomes / Q000821:virology; D008854:Microscopy, Electron; D012401:Rotavirus / Q000254:growth & development*; D014771:Virion / Q000648:ultrastructure
chemical_list: D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone