Delineation of protective epitopes on the E2-envelope glycoprotein of Semliki Forest virus.
Two short linear peptides, 17 and 14 amino acids long, on the Semliki Forest virus (SFV) E2-envelope polypeptide are shown to be involved in the protection of mice against lethal challenge with SFV. Peptides corresponding to these two regions, designated H and L, were selected for study on the basis of our model for prediction of protective epitopes on E2 polypeptide of alphaviruses. These peptides were produced in Escherichia coli as recombinant proteins fused to the amino terminus of beta-galactosidase. Both the H epitope (amino acid positions 227-243 on E2) and L epitope (amino acid positions 297-310) are recognized by antibodies raised against SFV, and both trigger antibodies that interact with native SFV-E2. Vaccination of mice with the H-beta-galactosidase polypeptide confers 64-87% protection against a lethal viral challenge (250 LD50), and immunization with L-beta-galactosidase leads to 23-66% protection of challenged mice. The efficacy of the L-based synthetic vaccine could be improved further (up to 100% protection) by presentation of this epitope as a dimer fused to beta-galactosidase. These results provide evidence that the algorithm and the methodology proposed by us previously are effective tools for identification of linear protective epitopes on E2-envelope of SFV.
Grosfeld
H
H
Department of Biochemistry, Israel Institute for Biological Research, Ness-Ziona.
Velan
B
B
Leitner
M
M
Lustig
S
S
Lachi
B E
BE
Cohen
S
S
Shafferman
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D005260:Female; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012672:Semliki forest virus / Q000235:genetics / Q000276:immunology*; D014036:Togaviridae Infections / Q000517:prevention & control; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000302:isolation & purification* / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics
D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; D004247:DNA; D001616:beta-Galactosidase