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Efficient identification of HLA-A*2402-restricted cytomegalovirus-specific CD8 1 T-cell epitopes by a computer algorithm and an enzyme-linked immunospot assay

Efficient identification of HLA-A*2402-restricted cytomegalovirus-specific CD8 1 T-cell epitopes by a computer algorithm and an enzyme-linked immunosp

Efficient identification of HLA-A*2402-restricted cytomegalovirus-specific CD8 1 T-cell epitopes by a computer algorithm and an enzyme-linked immunospot assay   (Citations: 32)
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Antigenic peptides recognized by virus- specific cytotoxic T lymphocytes (CTLs) are useful tools for studying the CTL responses exclusively among those who own the major histocompatibility com- plex (MHC) class I molecules that present the peptides. For widening the applica- tion, an efficient strategy to determine such epitopes in the context of a given MHC is highly desirable. A rapid and efficient strategy is presented for the determination of CTL epitopes in the context of given MHC molecules of inter- est through multiple screenings consist- ing of a computer-assisted algorithm and MHC stabilization and enzyme-linked immunospot assays. A major cytomega- lovirus (CMV)-specific CTL epitope, QYDPVAALF, in the amino acid sequence of its lower matrix 65 kd phosphoprotein (pp65) presented by HLA-A*2402 mol- ecules was identified from 83 candidate peptides. The results indicate that the CMV-specific CTL response is highly fo- cused to pp65 in the context of HLA- A*2402. Endogenous processing and pre- sentation was confirmed using a peptide- specific CD81 T-cell clone as the effectors and autologous fibroblast cells infected with recombinant vaccinia virus express- ing pp65 gene or CMV as antigen-present- ing cells. Flow cytometric analysis of intracellular interferon-g production re- vealed 0.04% to 0.27% of CD81 T cells in peripheral blood of HLA-A241 and CMV- seropositive donors to be specific for the peptide. The tetrameric MHC-peptide com- plexes specifically bound to the reactive T-cell clone and 0.79% of CD81 T cells in peripheral blood from a seropositive do- nor. The peptide could be a useful reagent to study CTL responses to CMV among populations positive for HLA-A*2402. (Blood. 2001;98:1872-1881)
Published in 2010.
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