Abstract
The S gene of transmissible gastroenteritis virus (TGEV) was inserted into the genome of Autographa californica nuclear polyhedrosis virus (AcNPV) using the transfer plasmid pVL941. Infection of Sf9 insect cells with the recombinant virus resulted in the synthesis of a 175K polypeptide which was able to trimerize and was transported to the cell surface as is the authentic TGEV S protein. Despite the lack of complete carbohydrate processing, the recombinant S protein exhibited antigenic properties similar to TGEV S and induced high levels of neutralizing antibodies in immunized rats. Engineering a deletion (70 amino acids) into the carboxy-terminus containing the membrane anchor of the polypeptide allowed its secretion. The oligomerization process and the antigenic profile of the anchor-free S protein were shown to be partially altered.
References
- Britton P., Page K.W. Sequence of the S gene from a virulent British field isolate of transmissible gastroenteritis virus. Virus Res. 1990;18:71–80. doi: 10.1016/0168-1702(90)90090-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Correa I., Jimenez G., Sune C., Bullido M.J., Enjuanes L. Antigenic structure of the E2 glycoprotein from transmissible gastroenteritis coronavirus. Virus Res. 1988;10:77–94. doi: 10.1016/0168-1702(88)90059-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Correa I., Gebauer F., Bullido M.J., Sune C., Baay M.F.D., Zwaagstra K.A., Posthumus W.P.A., Lenstra J.A., Enjuanes L. Localization of antigenic sites of the E2 glycoprotein of transmissible gastroenteritis coronavirus. J. Gen. Virol. 1990;71:271–279. doi: 10.1099/0022-1317-71-2-271. [DOI] [PubMed] [Google Scholar]
- De Groot R.J., Luytjes W., van Der Zeijst B.A.M., Spaan W.J.M., Lenstra J.A. Evidence for a coiled-coil structure in the spike proteins of coronaviruses. J. Mol. Biol. 1987;196:963–966. doi: 10.1016/0022-2836(87)90422-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Delmas B., Gelfi J., Laude H. Antigenic structure of transmissible gastroenteritis virus. II. Domains in the peplomer glycoprotein. J. Gen. Virol. 1986;67:1405–1418. doi: 10.1099/0022-1317-67-7-1405. [DOI] [PubMed] [Google Scholar]
- Delmas B., Laude H. Assembly of coronavirus spike protein into trimers and its role in epitope expression. J. Virol. 1990;64:5367–5375. doi: 10.1128/jvi.64.11.5367-5375.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Delmas B., Laude H. Carbohydrate-induced conformational changes strongly modulate the antigenicity of coronavirus TGEV glycoproteins S and M. Virus Res. 1991;20:107–120. doi: 10.1016/0168-1702(91)90103-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Delmas B., Rasschaert D., Godet M., Gelfi J., Laude H. Four major antigenic sites of the coronavirus transmissible gastroenteritis virus are located on the amino-terminal half of spike glycoprotein S. J. Gen. Virol. 1990;71:1313–1323. doi: 10.1099/0022-1317-71-6-1313. [DOI] [PubMed] [Google Scholar]
- Domingo D.L., Trowbridge I.S. Characterization of the human transferrin receptor produced in a baculovirus expression system. J. Biol. Chem. 1988;263:13386–13392. [PubMed] [Google Scholar]
- Doms R.W., Helenius A. Quaternary structure of influenza virus hemagglutinin after acid treatment. J. Virol. 1986;60:833–839. doi: 10.1128/jvi.60.3.833-839.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garwes D.J., Lucas M.H., Higgins D.A., Pike B.V., Cartwright S.F. Antigenicity of structural components from porcine transmissible gastroenteritis virus. Vet. Microbiol. 1978/1979;3:179–190. [Google Scholar]
- Jimenez G., Correa I., Melgosa M.P., Bullido M.J., Enjuanes L. Critical epitopes in transmissible gastroenteritis virus neutralization. J. Virol. 1986;60:131–139. doi: 10.1128/jvi.60.1.131-139.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu. Rev. Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
- Kuroda K., Geyer H., Geyer R., Doerfler W., Klenk H.D. The oligosaccharides of influenza virus hemagglutinin expressed in insect cells by a baculovirus vector. Virology. 1990;174:418–429. doi: 10.1016/0042-6822(90)90095-9. [DOI] [PubMed] [Google Scholar]
- Kuroda K., Veit M., Klenk H.D. Retarded processing of influenza virus hemagglutinin in insect cells. Virology. 1991;180:159–165. doi: 10.1016/0042-6822(91)90019-8. [DOI] [PubMed] [Google Scholar]
- Laude H., Chapsal J.M., Gelfi J., Labial S., Grosclaude J. Antigenic structure of transmissible gastroenteritis virus. I. Properties of monoclonal antibodies directed against virion proteins. J. Gen. Viol. 1986;67:119–130. doi: 10.1099/0022-1317-67-1-119. [DOI] [PubMed] [Google Scholar]
- Lim H., Pène J.J. Optimal conditions for supercoil DNA sequencing with the Escherichia coli DNA polymerase I large fragment. Gene Anal. Tech. 1988;5:32–39. doi: 10.1016/0735-0651(88)90024-6. [DOI] [PubMed] [Google Scholar]
- Luckow V.A., Summers M.D. High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors. Virology. 1989;170:31–39. doi: 10.1016/0042-6822(89)90348-6. [DOI] [PubMed] [Google Scholar]
- Posthumus W.A., Lenstra J.A., Schaaper W.M.M., van Nieuw-Stadt A.P., Enjuanes L., Meloen R.H. Analysis and simulation of a neutralizing epitope of transmissible gastroenteritis virus. J. Virol. 1990;64:3304–3309. doi: 10.1128/jvi.64.7.3304-3309.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rasschaert D., Laude H. The predicted structure of the peplomer protein E2 of the porcine coronavirus primary transmissible gastroenteritis virus. J. Gen. Virol. 1987;68:1883–1890. doi: 10.1099/0022-1317-68-7-1883. [DOI] [PubMed] [Google Scholar]
- Rasschaert D., Gelfi J., Laude H. Enteric coronavirus TAEV: Partial sequence of the genomic RNA, its organization and expression. Biochimie. 1987;69:591–600. doi: 10.1016/0300-9084(87)90178-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rasschaert D., Duarte M., Laude H. Porcine respiratory coronavirus differs from transmissible gastroenteritis virus by a few genomic deletions. J. Gen. Virol. 1990;71:2599–2607. doi: 10.1099/0022-1317-71-11-2599. [DOI] [PubMed] [Google Scholar]
- Scharf S.J., Horn G.T., Erlich H.A. Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science. 1986;233:1076–1078. doi: 10.1126/science.3461561. [DOI] [PubMed] [Google Scholar]
- Spaan W., Cavanagh D., Horzinek M.C. Coronaviruses: structure and genome expression. J. Gen. Virol. 1988;69:2939–2952. doi: 10.1099/0022-1317-69-12-2939. [DOI] [PubMed] [Google Scholar]
- Summers M.D., Smith G.E. Texas agricultural Station; College Station, TX: 1987. A manual of methods for baculovirus vectors and insect cell culture procedure. (Texas Agricultural Experiment Station bulletin No. 1555). [Google Scholar]
- Van-Drunen Little-van Den Hurk S., Parker M.D., Fitzpatrick D.R., Zamb T.J., van Den Hurk J.V., Campos M., Harland R., Babiuk L.A. Expression of bovine Herpesvirus 1 glycoprotein glV by recombinant baculovirus and analysis of its immunogenic properties. J. Virol. 1991;65:263–271. doi: 10.1128/jvi.65.1.263-271.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takase-Yoden S., Kikuchi T., Siddell S.G., Taguchi F. Localization of major neutralizing epitopes on the S1 polypeptide of the murine coronavirus peplomer glycoprotein. Virus Res. 1990;18:99–108. doi: 10.1016/0168-1702(91)90011-J. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wesley R.D. Nucleotide sequence of the E2-peplomer protein gene and partial nucleotide sequence of the upstream polymerase gene of transmissible gastroenteritis virus (Miller strain) In: Cavanagh D., Brown T.D.K., editors. Coronaviruses and Their Diseases. Vol. 276. Plenum Press; New York: 1990. (Adv. Exp. Med. Biol.). [DOI] [PubMed] [Google Scholar]
- Yoden S., Kikuchi T., Siddell S.G., Taguchi F. Expression of the peplomer glycoprotein of murine coronavirus JHM using a baculovirus vector. Virology. 1989;173:615–623. doi: 10.1016/0042-6822(89)90573-4. [DOI] [PMC free article] [PubMed] [Google Scholar]