Oral Presentation Australian Society for Microbiology Annual Scientific Meeting 2013

Quantitative kinetics of antigen presentation during vaccinia virus infection (#124)

David C Tscharke 1 , Nathan P Croft 2 , Stewart A Smith 1 , Yik Chun Wong 1 , Leon Lin 1 , Inge EA Flesch 1 , Chor Teck Tan 2 , Nadine Dudek 2 , Anthony Purcell 2
  1. Research School of Biology, The Australian National University, Canberra, ACT, Australia
  2. Department of Biochemistry and Molecular Biology, Monash University, Clayton, Vic, Australia
CD8+ T cell responses to viral infection are driven by the presentation of peptide epitopes on the surface of cells in the context of major histocompatibility (MHC) class I complexes. Despite a thorough understanding of MHC antigen processing and presentation during viral infection, there is a lack of knowledge relating to the relative kinetics of MHC-peptide presentation from across a viral proteome. This is especially relevant for large viruses with ordered cascades of gene expression. In this context vaccinia virus (VACV) is an ideal model, with a well characterised CD8+ T cell epitope hierarcy in mice that includes antigens from all kinetic classes of virus genes. The abundance of 17 VACV MHC-peptide complexes were quantified in parallel using mass spectrometric methods at multiple times over 12 hours of infection. Furthermore, levels of source proteins for each peptide were simultaneously measured and related to MHC-peptide levels. Finally, the biochemical data were related to the known immunodomince of the epitopes. Our results represent the first multi-epitope study quantifying MHC presentation of viral epitopes during infection and provide important insights into the kinetics of viral epitope display. Further, they show that antigen presentation during VACV infection is more complex than has been suggested by previous studies and that there is no simple relationship between peptide presentation levels and immunodominance across that viral proteome.