High-level expression of the measles virus nucleocapsid protein by using a replication-deficient adenovirus vector: induction of an MHC-1-restricted CTL response and protection in a murine model.

Abstract:

:Replication-deficient adenovirus (Ad) vectors provide an efficient technology for direct DNA delivery to cells both in vitro and in vivo. We have inserted the measles virus nucleoprotein (N) gene under the control of the strong constitutive CMV major IE promoter into an Ad type 5 E1- vector to produce the recombinant virus RAd68. Following infection of human fibroblasts with RAd68 in vitro, recombinant N protein was synthesized as a 60-kDa protein that represented up to 20% total soluble cell protein. Long filamentous structures were produced in both the nucleus and the cytoplasm that were similar in appearance to measles virus nucleocapsids. These "nucleocapsid-like" structures were readily purified by density gradient centrifugation. Murine immunization with RAd68 elicited (i) a humoral immune response to N, (ii) a major histocompatibility complex class I-restricted, antigen-specific cytotoxic T cell response, and (iii) protection against challenge with the measles virus CAM/RB strain in mice. This study demonstrates the capacity of replication-deficient Ad recombinants both to induce and to characterize cell-mediated immune responses.

journal_name

Virology

journal_title

Virology

authors

Fooks AR,Schadeck E,Liebert UG,Dowsett AB,Rima BK,Steward M,Stephenson JR,Wilkinson GW

doi

10.1006/viro.1995.1362

subject

Has Abstract

pub_date

1995-07-10 00:00:00

pages

456-65

issue

2

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(85)71362-1

journal_volume

210

pub_type

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