Abstract: Viral CD8 T-cell epitopes, represented by viral peptides bound to major histocompatibility complex class-I (MHC-I) glycoproteins, are often identified by “reverse immunology”, a strategy not requiring biochemical and structural knowledge of the actual viral protein from which they are derived by antigen processing. Instead, bioinformatic algorithms predicting the probability of C-terminal cleavage in the proteasome, as well as binding affinity to the presenting MHC-I molecules, are applied to amino acid sequences deduced from predicted open reading frames (ORFs) based on the genomic sequence. If the protein corresponding to an antigenic ORF is known, it is usually inferred that the kinetic class of the protein also defines the phase in the viral replicative cycle during which the respective antigenic peptide is presented for recognition by CD8 T cells. We have previously identified a nonapeptide from the predicted ORFm164 of murine cytomegalovirus that is presented by the MHC-I allomorph H-2 Dd and that is immunodominant in BALB/c (H-2d haplotype) mice. Surprisingly, although the ORFm164 protein gp36.5 is expressed as an Early (E) phase protein, the m164 epitope is presented already during the Immediate Early (IE) phase, based on the expression of an upstream mRNA starting within ORFm167 and encompassing ORFm164.
Keywords: antigenic peptides; antigen presentation; CD8 T cell epitope; gene expression; immediate-early protein; murine CMV ORF m164; open reading frame; RACE mapping; reverse immunology; transcription start site; translation start site
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Fink, A.; Büttner, J.K.; Thomas, D.; Holtappels, R.; Reddehase, M.J.; Lemmermann, N.A.W. Noncanonical Expression of a Murine Cytomegalovirus Early Protein CD8 T-Cell Epitope as an Immediate Early Epitope Based on Transcription from an Upstream Gene. Viruses 2014, 6, 808-831.
Fink A, Büttner JK, Thomas D, Holtappels R, Reddehase MJ, Lemmermann NAW. Noncanonical Expression of a Murine Cytomegalovirus Early Protein CD8 T-Cell Epitope as an Immediate Early Epitope Based on Transcription from an Upstream Gene. Viruses. 2014; 6(2):808-831.
Fink, Annette; Büttner, Julia K.; Thomas, Doris; Holtappels, Rafaela; Reddehase, Matthias J.; Lemmermann, Niels A.W. 2014. "Noncanonical Expression of a Murine Cytomegalovirus Early Protein CD8 T-Cell Epitope as an Immediate Early Epitope Based on Transcription from an Upstream Gene." Viruses 6, no. 2: 808-831.