title: High-level expression of the tick-borne encephalitis virus NS1 protein by using an adenovirus-based vector: protection elicited in a murine model.
abstract: Tick-borne encephalitis virus (TBEV) encodes an abundant, highly immunogenic nonstructural glycoprotein, NS1. The function of this protein has yet to be determined. We have cloned the NS1 gene from the Neudorfl strain of TBEV under the control of the powerful constitutive cytomegalovirus major immediate-early promoter into an adenovirus E1 deletion mutant. The novel combination of the cytomegalovirus immediate-early promoter and the adenovirus vector produced extremely high levels of NS1 expression in cells which do not support replication of the adenovirus deletion mutant. The recombinant protein was shown to be indistinguishable from authentic TBEV NS1 in its (i) apparent molecular weight by polyacrylamide gel electrophoresis, (ii) glycosylation pattern, (iii) ability to form high-molecular-weight complexes, and (iv) ability to be secreted from cells. Appropriate processing of NS1 expressed by the adenovirus recombinant occurred independently of any additional TBEV-encoded gene function. When directly inoculated into mice, the recombinant adenovirus RAd51 was shown to elicit an antibody response to the TBEV NS1 protein. Immunization with RAd51 conferred protection against challenge with TBEV.
authors.affiliation: Biologics Division, Centre for Applied Microbiology and Research, Porton Down, Salisbury, United Kingdom.
authors.forename: S C
authors.initials: SC
authors.lastname: Jacobs
authors.forename: J R
authors.initials: JR
authors.lastname: Stephenson
authors.forename: G W
authors.initials: GW
authors.lastname: Wilkinson
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D004669:Encephalitis Viruses, Tick-Borne / Q000235:genetics*; D004675:Encephalitis, Tick-Borne / Q000517:prevention & control*; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D007114:Immunization; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines / Q000008:administration & dosage
chemical_list: D004279:DNA, Viral; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines