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Journal of Virology, October 2002, p. 10138-10146, Vol. 76, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.20.10138-10146.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Newcastle Disease Virus (NDV) Marker Vaccine: an Immunodominant Epitope on the Nucleoprotein Gene of NDV Can Be Deleted or Replaced by a Foreign Epitope

Teshome Mebatsion,1* Marck J. M. Koolen,1 Leonie T. C. de Vaan,1 Niels de Haas,1 Marian Braber,1 Angela Römer-Oberdörfer,2 Paul van den Elzen,1 and Pieter van der Marel1

Department of Virology, Intervet International B.V., 5830 AA Boxmeer, The Netherlands,1 Institute of Molecular Biology, Friedrich-Loeffler Institute, Federal Research Centre for Virus Diseases of Animals, D-17498 Insel Riems, Germany2

Received 6 May 2002/ Accepted 17 July 2002

The nucleoprotein (NP) of Newcastle disease virus (NDV) functions primarily to encapsidate the virus genome for the purpose of RNA transcription, replication, and packaging. This conserved multifunctional protein is also efficient in inducing NDV-specific antibody in chickens. Here, we localized a conserved B-cell immunodominant epitope (IDE) spanning residues 447 to 455 and successfully generated a recombinant NDV lacking the IDE by reverse genetics. Despite deletion of NP residues 443 to 460 encompassing the NP-IDE, the mutant NDV propagated in embryonated specific-pathogen-free chicken eggs to a level comparable to that of the parent virus. In addition, a B-cell epitope of the S2 glycoprotein of murine hepatitis virus (MHV) was inserted in-frame to replace the NP-IDE. Recombinant viruses properly expressing the introduced MHV epitope were successfully generated, demonstrating that the NP-IDE not only is dispensable for virus replication but also can be replaced by foreign sequences. Chickens immunized with the hybrid recombinants produced specific antibodies against the S2 glycoprotein of MHV and completely lacked antibodies directed against the NP-IDE. These marked-NDV recombinants, in conjunction with a diagnostic test, enable serological differentiation of vaccinated animals from infected animals and may be useful tools in ND eradication programs. The identification of a mutation-permissive region on the NP gene allows a rational approach to the insertion of protective epitopes and may be relevant for the design of NDV-based cross-protective marker vaccines.

* Corresponding author. Mailing address: Department of Virology, Intervet International B.V., P.O. Box 31, 5830 AA Boxmeer, The Netherlands. Phone: 31 485 587 351. Fax: 31 485 585 317. E-mail: teshome.mebatsion{at}intervet.com.

Journal of Virology, October 2002, p. 10138-10146, Vol. 76, No. 20
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.20.10138-10146.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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