MHC Associated Peptide Proteomics
In Vitro System To Identify Presented Epitopes That Induce Cytotoxic CD8+ T Cell Responses
By comparing the MHC class I presented peptides of viral infected cells & tumour cells with equivalent healthy cells using MAPPs, utilising powerful mass spectrometry, allows for the identification of antigens or neo-antigens for therapeutic targets. Combining in vitro T cell assays allows for confirmation of whether identified peptides isolated from MHC Class I molecules contain epitopes that can elicit CD8+ T cell response. Relative quantification and affinity of peptides that elicit T cell response can be measured by MAPPs.
The primary immune response to an antigen is initiated through antigen presenting cells such as dendritic cells (DC). Human leukocyte antigens (HLA) on the surface of these cells present protein-derived peptide fragments, where MHC class I associated HLA (i.e. HLA-A, B and C) present peptides to CD8+ T cells and MHC class II associated HLA (HLA-DR, DP and DQ) to CD4+ T cells. MHC-I presentation is fundamental for the development of immunity against tumours and viruses since endogenous proteins are normally presented to CD8+ T cells via this route. Presentation of exogenous antigens via MHC class I can also occur and trigger an immune response. Therefore, the identification of these antigens presented via MHC class I creates huge potential for the generation of new immuno-oncology therapeutics, vaccines and diagnostics.
To identify potential antigens for vaccine design putative antigens (i.e. viral proteins, tumour associated antigens) are expressed in human cell lines for a comparative analysis of the presented peptides. Unique viral peptides identified can be synthesised for use in in vitro T cell assays to assess for CD8+ T cell activation.
Tumour Antigen Discovery & Validation
MAPPs can identify tumour associated antigens or neo-antigens based on comparisons with healthy tissue. Any variable presented protein peptides can be synthesised to check for CD8+ T cell activation. This has huge potential for the discovery of new targets for immunotherapeutics.
T Cell responses can be associated with disease outcome (i.e. chronic infection, cancer). Through MAPPs relevant T cell epitopes can be identified from whole proteins and tumour antigens and used in assay design to monitor effective T cell responses ex vivo from patients.
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