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1、Vincr Research, 16 (1990) 185-194 Ekevier 185 VIRUS 00580 Hemagglutinating encephalomyelitis virus attaches to IV-acetyl-9-O-acetylneuraminic acid-containing receptors on erythrocytes: comparison with bovine coronavirus and influenza C virus Beate Schultze , Hans-Jiirgen Gross *, Reinhard Brossmer *
2、, Hans-Dieter Klenk and Georg Hkrrler I Institut fiir Virologie, Philipps- Universitiir Marburg Marburg F. R. G. and 2 Institut ftir Biochemie II, Universitiit Heidelberg, Heidelberg, F. R. G. (Accepted 20 February 1990) The receptors for the hemagglutinating encephalomyelitis virus (HEV, a porcine
3、coronavirus) on chicken erythrocytes were analyzed and compared to the receptors for bovine coronavirus (BCV) and influenza C virus. Evidence was obtained that HEV requires the presence of N-acetyl-9-0-acetylneuraminic acid (Neu5,9Ac,) on the cell surface for agglutination of erythrocytes as has bee
4、n previously shown for BCV and influenza C virus: (i) Incubation of red blood cells with sialate 9-0- acetylesterase, the receptor-destroying enzyme of influenza C virus, rendered the erythrocytes resistant against agglutination by each of the three viruses; (ii) Human erythrocytes which are resista
5、nt to agglutination by HEV acquire receptors for HEV after resialylation with Neu5,9Ac,. Sialylation of red blood cells with limiting amounts of sialic acid indicated that strain JHB/1/66 of influenza C virus requires less Neu5,9Ac, for agglutination of erythrocytes than the two coronaviruses, both
6、of which were found to be similar in their reactivity with Neu5,9Ac,-containing receptors. Bovine coronavirus; Hemagglutinating encephalomyelitis virus; receptor; Sialic acid; N-acetyl-9-0-acetyhreuraminic acid Correspondence to: G. Herder, Institut fiir Virologie, Robert-Koch-Str. 17, D-3550 Marbur
7、g, F.R.G. 0168-1702/90/%03.50 6 1990 Ekevier Science Publishers B.V. (Biomedical Division) 186 Introduction There are great variations among members of the family Coronaviridae in their ability to agglutinate red blood cells. Some strains such as A59 or JHM of mouse hepatitis virus are completely de
8、void of hemagglutinating activity. Other coronaviruses have been reported to be poor hemagglutinins, for instance transmis- sible gastroenteritis virus (Noda et al., 1987) or infectious bronchitis virus (Bingham et al., 1975). Several members of this virus family, however, are quite potent hemagglut
9、inating agents. These include bovine coronavirus (BCV), human coronavirus 0C43 (HCV-OC43), hemagglutinating encephalomyelitis virus (HEV), and some murine coronaviruses (Kaye and Dowdle, 1969; Pensaert and Callebaut, 1974; Sato et al., 1977; Sugiyama and Amano, 1980). Information about the nature of
10、 the erythrocyte receptors recognized by these viruses became available only recently, when BCV was reported to contain a sialate 9-0-acetylesterase (Vlasak et al., 1988a). The same enzyme, which releases 9-0-acetyl residues from sialic acid, has previously been shown to be present on influenza C vi
11、rions and to inactivate cellular receptors for influenza C virus (Herrler et al., 1985; Herrler and Klenk, 1987). These findings indicated that N-acetyl-9-0-acetylneuraminic acid (Neu59Ac,) is a receptor determinant for attachment of influenza C virus to cells, which was confirmed by resialylation s
12、tudies (Rogers et al., 1986). The esterases of both BCV and influenza C virus have been shown to inactivate the receptors on erythrocytes for BCV as well as for HCV-0C43 (Vlasak et al., 1988a). This result indicates that Neu5,9Ac, is a receptor determinant for attachment of BCV and HCV-OC43 to eryth
13、rocytes. Here we report that the hemagglutinating activity of HEV is also dependent on the presence of NeuSQAc, on the erythrocyte surface. Evidence is presented that strain JHB/1/66 of influenza C virus is more efficient in recognizing Neu5,9Ac,- containing receptors than HEV and BCV, which both re
14、quire similar amounts of 9-0-acetylated sialic acid on the cell surface in order to cause hemagglutination. Materials and Methods Viruses Strain NT-9 of HEV was obtained from Dr R.G. Hess (Koblenz, F.R.G.). Strain L9 of BCV was provided by Dr R. Rott (Giessen, F.R.G.). Both coronaviruses were grown
15、in MDCK I cells, a subline of Madin-Darby canine kidney cells, which were maintained as described previously (Herrler et al., 1988). After an adsorption period of 60 min at 37”C, the inoculum was removed, and the infected cells were incubated at 37 o C with serum-free Eagles minimal essential medium
16、 (MEM) in the presence of trypsin (1 pg/ml). Virus was harvested 48 h p.i. The supernatant, which usually had an HA-titer of 256-512 HA-units/ml, was clarified by low speed centrifugation and, following addition of fetal calf serum (lo%), stored at -80 o C. 187 Strain Johannesburg/l/66 (JHB/1/66) of influenza C virus and fowl plague virus (influenza A/FPV/Rostock/34 (H7Nl) were grown in embryonated eggs as described previously (Herrler and Klenk, 1987; Klenk et al., 1972).
