Identification of a 25-aminoacid sequence from the major African swine fever virus structural protein VP72 recognised by porcine cytotoxic T lymphocytes using a lipoprotein based expression system.
Identification of African swine fever virus (ASFV) proteins recognised by cytotoxic T lymphocytes (CTL) from swine surviving ASFV/NH/P68 infection was assessed using expression vectors based on the Pseudomonas aeruginosa outer membrane lipoprotein I gene (oprI). Viral antigens expressed as fusion lipoproteins were shown to be taken efficiently by porcine blood-derived macrophages incubated with outer membrane protein preparations from transformed E. coli. To assess recognition by CTL the fusion lipoprotein-treated macrophages were used as targets in 51Cr release microcytotoxicity assays. Using this approach it was shown that the aminoacid sequence HKPHQSKPILTDENDTQRTCSHTNP from the major structural ASFV protein (VP72), encoded by a recombinant clone (pVUB72) is presented by macrophages, which are lysed under restriction of SLA class I antigens. Overall, the results demonstrate that the oprI based vectors are valuable tools to study ASFV-specific CTL activity.
Leitão
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Laboratório de Doenças Infecciosas, CIISA, Faculdade de Medicina Veterinária, Lisboa, Portugal.
Malur
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Cornelis
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Martins
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D000358:African Swine Fever Virus / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D015870:Gene Expression; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013552:Swine; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D001426:Bacterial Proteins; D036022:Capsid Proteins; D008074:Lipoproteins; D011993:Recombinant Fusion Proteins; C063745:capsid protein p72, African swine fever virus; C060507:outer membrane lipoprotein I, Bacteria