High-Throughput Stability Screening of Neoantigen/HLA Complexes Improves Immunogenicity Predictions

Cancer Immunol Res. 2019 Jan;7(1):50-61. doi: 10.1158/2326-6066.CIR-18-0395. Epub 2018 Nov 13.

Abstract

Mutated peptides (neoantigens) from a patient's cancer genome can serve as targets for T-cell immunity, but identifying which peptides can be presented by an MHC molecule and elicit T cells has been difficult. Although algorithms that predict MHC binding exist, they are not yet able to distinguish experimental differences in half-lives of the complexes (an immunologically relevant parameter, referred to here as kinetic stability). Improvement in determining actual neoantigen peptide/MHC stability could be important, as only a small fraction of peptides in most current vaccines are capable of eliciting CD8+ T-cell responses. Here, we used a rapid, high-throughput method to experimentally determine peptide/HLA thermal stability on a scale that will be necessary for analysis of neoantigens from thousands of patients. The method combined the use of UV-cleavable peptide/HLA class I complexes and differential scanning fluorimetry to determine the Tm values of neoantigen complexes. Measured Tm values were accurate and reproducible and were directly proportional to the half-lives of the complexes. Analysis of known HLA-A2-restricted immunogenic peptides showed that Tm values better correlated with immunogenicity than algorithm-predicted binding affinities. We propose that temperature stability information can be used as a guide for the selection of neoantigens in cancer vaccines in order to focus attention on those mutated peptides with the highest probability of being expressed on the cell surface.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antigens, Neoplasm / chemistry
  • Antigens, Neoplasm / immunology*
  • HLA-A2 Antigen / chemistry
  • HLA-A2 Antigen / immunology*
  • High-Throughput Screening Assays
  • Peptides / chemistry
  • Peptides / immunology*
  • Protein Stability
  • Temperature

Substances

  • Antigens, Neoplasm
  • HLA-A2 Antigen
  • Peptides