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Journal of Virology, May 2002, p. 4241-4250, Vol. 76, No. 9
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.9.4241-4250.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of Neutralizing and Nonneutralizing Epitopes in the Porcine Reproductive and Respiratory Syndrome Virus GP5 Ectodomain

M. Ostrowski,1 J. A. Galeota,2 A. M. Jar,3 K. B. Platt,4 F. A. Osorio,2 and O. J. Lopez1*

Wolf Biotech,1 University of Nebraska, Lincoln, Nebraska,2 University of Buenos Aires, Buenos Aires, Argentina,3 Iowa State University, Ames, Iowa4

Received 24 September 2001/ Accepted 29 January 2002

After infection of swine with porcine reproductive and respiratory syndrome virus (PRRSV), there is a rapid rise of PRRSV-specific nonneutralizing antibodies (NNA), while neutralizing antibodies (NA) are detectable not sooner than 3 weeks later. To characterize neutralizing epitopes, we selected phages from a 12-mer phage display library using anti-PRRSV neutralizing monoclonal antibody (MAb) ISU25-C1. In addition, phages carrying peptides recognized by swine antibodies with high seroneutralizing titer were isolated after subtracting from the library those clones binding to swine anti-PRRSV serum with no neutralizing activity. Two epitopes located in the ectodomain of PRRSV GP5 were identified. One of these epitopes, which we named epitope B, was recognized both by neutralizing MAb ISU25-C1 and swine neutralizing serum (NS) but not by swine nonneutralizing serum (NNS), indicating that it is a neutralizing epitope. Epitope B is sequential, conserved among isolates, and not immunodominant. Antibodies directed against it are detected in serum late after infection. In contrast, the other epitope, which we named epitope A, is hypervariable and immunodominant. Antibodies against it appear early after infection with PRRSV. This epitope is recognized by swine NNA but is not recognized by either neutralizing MAb ISU25-C1 or swine NA, indicating that it is not involved in PRRSV neutralization. During infection with PRRSV, epitope A may act as a decoy, eliciting most of the antibodies directed to GP5 and delaying the induction of NA against epitope B for at least 3 weeks. These results are relevant to the design of vaccines against PRRSV.

* Corresponding author. Mailing address: Wolf Biotech, 1921 Pinedale Ave., Lincoln, NE. Phone: (402) 472-6171. Fax: (402) 472-9690. E-mail: biowolf{at}inebraska.com.

Journal of Virology, May 2002, p. 4241-4250, Vol. 76, No. 9
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.9.4241-4250.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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