A new approach to epitope confirmation by sampling effector/memory T cells migrating to the lung.
The identification of T cell epitopes that elicit a weak T cell response is technically challenging due to the relatively low resolution of many available screening assays. Peptide immunization can confirm the immunogenicity of a given peptide, however, this also often induces a low frequency response in lymphoid organs. In this report, we use the murine gammaherpesvirus-68 model system to describe a novel technique to enrich for antigen-experienced T cells in vivo as an aid to epitope mapping. Mice are immunized with peptides containing putative epitopes then effector/memory cells which migrate to the lungs are washed out and tested for specificity using intracellular staining for interferon gamma (IFN-gamma). We show that the lung is a site where there are elevated numbers of antigen-experienced T cells and this can be exploited to confirm otherwise low frequency T cell responses. In addition, we identify two novel T cell epitopes in the ORF65 protein of MHV-68 which will be valuable tools in the dissection of the immune response to this model gammaherpesvirus.
Usherwood
Edward J
EJ
Department of Microbiology and Immunology, Dartmouth Medical School, 1 Medical Center Drive, Lebanon, NH 03755, USA. Edward.J.Usherwood@Dartmouth.edu
D000818:Animals; D000956:Antigens, Viral / Q000737:chemistry / Q000276:immunology*; D002465:Cell Movement; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D005260:Female; D018142:Gammaherpesvirinae / Q000276:immunology; D006566:Herpesviridae Infections / Q000276:immunology; D007156:Immunologic Memory; D007371:Interferon-gamma / Q000032:analysis; D008168:Lung / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D010455:Peptides; D007371:Interferon-gamma; D002772:Cholera Toxin