Identification of an atypical CD8 T cell epitope encoded by murine cytomegalovirus ORF-M54 gaining dominance after deletion of the immunodominant antiviral CD8 T cell specificities.
Identifieur interne : 001667 ( PubMed/Corpus ); précédent : 001666; suivant : 001668Identification of an atypical CD8 T cell epitope encoded by murine cytomegalovirus ORF-M54 gaining dominance after deletion of the immunodominant antiviral CD8 T cell specificities.
Auteurs : Rafaela Holtappels ; Niels A W. Lemmermann ; Doris Thomas ; Angélique Renzaho ; Matthias J. ReddehaseSource :
- Medical microbiology and immunology [ 1432-1831 ] ; 2015.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Antigens, Viral (chemistry), Antigens, Viral (immunology), CD8-Positive T-Lymphocytes (immunology), Computational Biology, Cytotoxicity, Immunologic, Epitope Mapping, Epitopes, T-Lymphocyte (chemistry), Epitopes, T-Lymphocyte (immunology), Female, Genome, Viral, Herpesviridae Infections (immunology), Herpesviridae Infections (virology), Histocompatibility Antigen H-2D (immunology), Immunodominant Epitopes (chemistry), Immunodominant Epitopes (immunology), Mice, Muromegalovirus (genetics), Muromegalovirus (immunology), Mutation, Open Reading Frames (genetics), Open Reading Frames (immunology), Peptide Library, Peptides (chemistry), Peptides (immunology).
- MESH :
- chemical , chemistry : Antigens, Viral, Epitopes, T-Lymphocyte, Immunodominant Epitopes, Peptides.
- chemical , immunology : Antigens, Viral, Epitopes, T-Lymphocyte, Histocompatibility Antigen H-2D, Immunodominant Epitopes, Peptides.
- genetics : Muromegalovirus, Open Reading Frames.
- immunology : CD8-Positive T-Lymphocytes, Herpesviridae Infections, Muromegalovirus, Open Reading Frames.
- virology : Herpesviridae Infections.
- Amino Acid Sequence, Animals, Computational Biology, Cytotoxicity, Immunologic, Epitope Mapping, Female, Genome, Viral, Mice, Mutation, Peptide Library.
Abstract
Control of murine cytomegalovirus (mCMV) infection is mediated primarily by CD8 T cells, with four specificities dominating in BALB/c mice. Functional deletion of the respective immunodominant epitopes (IDEs) in mutant virus Δ4IDE revealed a still efficient control of infection. In a murine model of hematopoietic cell transplantation and infection with Δ4IDE, an mCMV-specific open reading frame (ORF) library screening assay indicated a strong CD8 T cell reactivity against the ORF-M54 product, the highly conserved and essential mCMV homolog of human CMV DNA polymerase UL54, which is a known inducer of in vivo protection against mCMV by DNA immunization. Applying bioinformatic algorithms for CD8 T cell epitope prediction, the top-scoring peptides were used to stimulate ex vivo-isolated CD8 T cells and to generate cytolytic T cell lines; yet, this approach failed to identify M54 epitope(s). As an alternative, a peptide library consisting of 549 10-mers with an offset of two amino acids (aa), covering the complete aa-sequence of the M54 protein, was synthesized and used for the stimulation. A region of 12 aa proved to encompass an epitope. An 'alanine walk' over this antigenic 12-mer and all possible 11-, 10- and 9-mers derived thereof revealed aa-residues critical for antigenicity, and terminal truncations identified the H-2D(d) presented 8-mer M5483-90 as the optimal epitope. An increased frequency of the corresponding CD8 T cells in the absence of the 4 IDEs indicated immunodomination by the IDE-specific CD8 T cells as a mechanism by which the generation of M54-specific CD8 T cells is inhibited after infection with wild-type mCMV.
DOI: 10.1007/s00430-015-0404-3
PubMed: 25805564
Links to Exploration step
pubmed:25805564Le document en format XML
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<front><div type="abstract" xml:lang="en">Control of murine cytomegalovirus (mCMV) infection is mediated primarily by CD8 T cells, with four specificities dominating in BALB/c mice. Functional deletion of the respective immunodominant epitopes (IDEs) in mutant virus Δ4IDE revealed a still efficient control of infection. In a murine model of hematopoietic cell transplantation and infection with Δ4IDE, an mCMV-specific open reading frame (ORF) library screening assay indicated a strong CD8 T cell reactivity against the ORF-M54 product, the highly conserved and essential mCMV homolog of human CMV DNA polymerase UL54, which is a known inducer of in vivo protection against mCMV by DNA immunization. Applying bioinformatic algorithms for CD8 T cell epitope prediction, the top-scoring peptides were used to stimulate ex vivo-isolated CD8 T cells and to generate cytolytic T cell lines; yet, this approach failed to identify M54 epitope(s). As an alternative, a peptide library consisting of 549 10-mers with an offset of two amino acids (aa), covering the complete aa-sequence of the M54 protein, was synthesized and used for the stimulation. A region of 12 aa proved to encompass an epitope. An 'alanine walk' over this antigenic 12-mer and all possible 11-, 10- and 9-mers derived thereof revealed aa-residues critical for antigenicity, and terminal truncations identified the H-2D(d) presented 8-mer M5483-90 as the optimal epitope. An increased frequency of the corresponding CD8 T cells in the absence of the 4 IDEs indicated immunodomination by the IDE-specific CD8 T cells as a mechanism by which the generation of M54-specific CD8 T cells is inhibited after infection with wild-type mCMV. </div>
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