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J Gen Virol 72 (1991), 557-565; DOI 10.1099/0022-1317-72-3-557
© 1991 Society for General Microbiology

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Identification of linear epitopes on Semliki Forest virus E2 membrane protein and their effectiveness as a synthetic peptide vaccine

Alies Snijders1, Barry J. Benaissa-Trouw1, Tom A. M. Oosterlaken1, Wouter C. Puijk2, Willem P. A. Posthumus2, Rob H. Meloen2, Wil A. M. Boere1, Janine D. Oosting1, Cornelis A. Kraaijeveld1 and Harm Snippe1

1 Eijkman-Winkler Laboratory of Medical Microbiology, Academic Hospital Utrecht, HP G04.614, Postbox 85500, 3508 GA Utrecht
and2 Central Veterinary Institute, Postbox 65, 8200 AB Lelystad, The Netherlands

Semliki Forest virus (SFV) infection of mice was used as a model to study the applicability of synthetic peptides containing only linear epitopes as viral vaccines. The identification of linear epitopes with vaccine potential on the E2 membrane protein of SFV was based on the binding of SFV-specific antibodies to a set of overlapping synthetic hexapeptides (Pepscan) representing the whole E2 amino acid sequence. The 14 available E2-specific monoclonal antibodies which were protective in vivo proved to be unsuitable for the identification of linear epitopes because they recognized only conformational epitopes, as indicated by their lack of reactivity with unfolded, reduced E2 protein on immunoblots. Three epitopes were detected with polyclonal anti-SFV serum at amino acid positions 135 to 141, 177 to 185 and 240 to 246 of the E2 protein. Synthetic peptides containing these epitopes were coupled to a carrier protein and tested as a vaccine. Mice immunized with the peptide containing amino acids 240 to 255 of protein E2 were protected against a challenge with virulent SFV but protection of mice immunized with the peptides containing amino acids 126 to 141 or 178 to 186 was only marginally better than that of controls. The prechallenge sera of most peptide-immunized mice reacted with SFV-infected cells but none of these sera neutralized the virus in vitro. However, protection of mice correlated well with SFV-specific antibody titre, suggesting antibody-mediated protection.

Received 8 October 1990; accepted 29 November 1990.


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Copyright © 1991 by the Society for General Microbiology.