An immunodominant cytotoxic T cell epitope on the VP7 rotavirus protein overlaps the H2 signal peptide.
C57BL/6 (H-2b) mice were primed with the bovine RF strain of rotavirus to study the induction of CD8+ cytotoxic T lymphocytes (CTLs). These rotavirus-specific CTLs were detected only after in vitro restimulation with the virus. Using a recombinant vaccinia virus we identified the RF VP7 protein as a major target of these CTLs. The response against this protein was obtained also after in vitro restimulation with simian SA11 and human WA strains of rotavirus. Using published Db and Kb allele-specific motifs to predict possible CTL epitopes in the RF VP7 protein, we synthesized and tested 18 predicted peptides of VP7. Only one peptide was able to sensitize target cells at a concentration below 5 x 10(-7) M. This CTL epitope was also induced by immunization with the RF VP7 expressed with a baculovirus vector, and was shown to be immunodominant by its capacity to inhibit, in an unlabelled target assay, the bulk response against cells infected with recombinant vaccinia virus expressing VP7. This CTL epitope overlaps the H2 signal peptide of the protein.
Franco
M A
MA
Laboratoire de Virologie et Immunologie Moléculaires INRA, C.R.J., Jouy-en-Josas, France.
Prieto
I
I
Labbé
M
M
Poncet
D
D
Borras-Cuesta
F
F
Cohen
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral; D016367:Baculoviridae / Q000235:genetics; D016827:CD8 Antigens / Q000276:immunology; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D003429:Cross Reactions; D006183:H-2 Antigens / Q000276:immunology*; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D021382:Protein Sorting Signals / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012401:Rotavirus / Q000235:genetics / Q000276:immunology*; D013045:Species Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014616:Vaccinia virus / Q000235:genetics
D000956:Antigens, Viral; D016827:CD8 Antigens; D036022:Capsid Proteins; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C050779:VP7 protein, Rotavirus