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1、VIROLOGY224, 345351 (1996) ARTICLE NO. 0540 SHORT COMMUNICATION A Murine and a Porcine Coronavirus Are Released from Opposite Surfaces of the Same Epithelial Cells J. W. A. ROSSEN, C. P. J. BEKKER, G. J. A. M. STROUS,* M. C. HORZINEK, G. S. DVEKSLER, K. V. HOLMES,1and P. J. M. ROTTIER2 Virology Divi
2、sion, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, and Institute of Biomembranes, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands; *Laboratory of Cell Biology, Medical School, Utrecht University, Heidelberglaan 100, AZU-H02.314, 3584 CX Utrecht,
3、The Netherlands; and Department of Pathology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799 Received April 23, 1996; accepted August 5, 1996 Epithelial cells are important target cells for coronavirus infection. Earlier we have shown that transmissible gastroent
4、eritis coronavirus (TGEV) and mouse hepatitis coronavirus (MHV) are released from different sides of porcine and murine epithelial cells, respectively. To study the release of these viruses from the same cells, we constructed a porcine LLC-PK1 cell line stably expressing the recombinant MHV receptor
5、 cDNA (LMR cells). The MHV and TGEV receptor glycoproteins were shown by immunofluorescence to appear at the surface of the cells and to be functional so that the cells were susceptible to both MHV and TGEV infection. Both coronaviruses entered polarized LMR cells only through the apical surface. Re
6、mark- ably, while the cells remained susceptible to TGEV for long periods, infectability by MHV decreased with time after plating of the cells onto filters. This was not due to a lack of expression of the MHV receptor, since this glycoprotein was still abundant on the apical surface of these cells.
7、TGEV and MHV appeared to exit LMR cells from opposite sides. Whereas TGEV was released preferentially at the apical membrane, MHV was released preferentially at the basolateral surface. These results show that vesicles containing the two coronaviruses are targeted differently in LMR cells. We propos
8、e that the viruses are sorted at the Golgi complex into different transport vesicles that carry information directing them to one of the two surface domains. The apical release of TGEV and the basolateral release of MHV might be factors contributing to the difference in virus spread found between TG
9、EV and MHV in their respective natural hosts, the former causing mainly a localized enteric infection, the latter spreading through the body to other organs.q 1996 Academic Press, Inc. Coronaviruses are enveloped, positive-strand RNA vi-vesicular stomatitis virus are released from the apical and bas
10、olateral surfaces, respectively, in both Fischerruses that infect humans and animals. Each virus has a narrow host range. The course of infection ranges fromrat thyroid (FRT) and Caco-2 cells, two togaviruses, Sindbis and Semliki Forest virus, bud from the apicalsubclinical to lethal, and the sympto
11、matology from respi- ratory and enteric disease (most commonly) to hepatitis,membrane of FRT cells, but from the basolateral mem- brane of Caco-2 cells (3). In all these cases buddingperitonitis, encephalomyelitis, and other syndromes. Pri- mary replication is often limited to epithelial cells of th
12、eappeared to take place at the plasma membrane domain where the respective viral membrane proteins wererespiratory or gastrointestinal tracts (1). The plasma membrane of epithelial cells is dividedfound to accumulate. Unlike the viruses mentioned above which mature byinto an apical domain, directed
13、to the external milieu, and a basolateral domain, facing the internal milieu. Manybudding from plasma membranes, coronaviruses mature by budding from intracytoplasmic membranes in a pre-viruses enter these cells from a specific side and also virus release is often vectorial (for a review, see Ref. 2
14、).Golgi compartment called the budding or intermediate compartment (46). From there the viral particles areThe polarity of virus release may differ in epithelial cells depending on their origin. Whereas influenza virus andtransported in vesicles through the secretory pathway to the plasma membrane,
15、where they are released by exocytosis (7). Recently, we have shown that the mouse 1Present address: Department of Microbiology, University of Colo- hepatitis coronavirus (MHV) is preferentially secreted rado Health Sciences Center, Campus Box B-175, 4200 East Ninth through the basolateral side of po
16、larized murine mTAL Avenue, Denver, Colorado 80262. cells, but that the porcine transmissible gastroenteritis 2To whom correspondence and reprint requests should be ad- dressed. Fax: *31-302536723; E-mail: P.Rottiervetmic.dgk.ruu.nl.coronavirus (TGEV) is secreted from apical surfaces of 345 0042-6822/96 $18.00 Copyright q 1996 by Academic Press, Inc. All rights of reproduction in any form reserved. AIDVY 8171/6a1f$82109-03-96 00:16:07virasAP: Virology 346SHORT COMMUNICATION FIG. 1. Susceptib
