Mapping of the target antigens of the rotavirus-specific cytotoxic T cell response.
The cytotoxic T cell (CTL) response in C57/B16 (H-2b) mice to rotavirus has been analysed using a cognate set of vaccinia virus recombinants covering the 12 primary gene products of the UKtc strain of bovine rotavirus. The gene products of RNA segments 5 (VP5/NSP-1) and 8 (VP7) both elicited a classic CD8+ Class I MHC restricted CTL response. Using L cells transfected with specific Class I MHC loci as targets the VP5/NSP-1 response was found to be restricted at Db and the VP7 response at Kb. Vaccinia virus recombinants expressing VP7 genes from seven G serotypes were used to show that the CTL response to this antigen is completely cross-reactive. By contrast, using the same strategy the CTL response to VP5/NSP-1 was found to be virus strain specific. A vaccinia virus recombinant carrying RNA segment 5 from the deletion mutant P9D delta 5 was used to localize at least one CTL epitope in VP5/ NSP-1 to the first 150 amino acids of the protein. The expression of a number of fragments of VP7 in vaccinia virus recombinants was used to show that the CTL epitope (amino acids 31-40) previously identified through the use of synthetic peptides is virus serotype specific rather than cross-reactive.
Heath
R R
RR
Department of Biological Sciences, University of Warwick, Coventry, UK.
Stagg
S
S
Xu
F
F
McCrae
M A
MA
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D002417:Cattle; D002460:Cell Line; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D015870:Gene Expression; D005822:Genetic Vectors; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D011995:Recombination, Genetic; D012401:Rotavirus / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014616:Vaccinia virus / Q000235:genetics
D000956:Antigens, Viral; D036022:Capsid Proteins; D018984:Epitopes, T-Lymphocyte; D011993:Recombinant Fusion Proteins; C050779:VP7 protein, Rotavirus