Pooled-Peptide Epitope Mapping Strategies Are Efficient and Highly Sensitive: An Evaluation of Methods for Identifying Human T Cell Epitope Specificities in Large-Scale HIV Vaccine Efficacy Trials

PLoS One. 2016 Feb 10;11(2):e0147812. doi: 10.1371/journal.pone.0147812. eCollection 2016.

Abstract

The interferon gamma, enzyme-linked immunospot (IFN-γ ELISpot) assay is widely used to identify viral antigen-specific T cells is frequently employed to quantify T cell responses in HIV vaccine studies. It can be used to define T cell epitope specificities using panels of peptide antigens, but with sample and cost constraints there is a critical need to improve the efficiency of epitope mapping for large and variable pathogens. We evaluated two epitope mapping strategies, based on group testing, for their ability to identify vaccine-induced T-cells from participants in the Step HIV-1 vaccine efficacy trial, and compared the findings to an approach of assaying each peptide individually. The group testing strategies reduced the number of assays required by >7-fold without significantly altering the accuracy of T-cell breadth estimates. Assays of small pools containing 7-30 peptides were highly sensitive and effective at detecting single positive peptides as well as summating responses to multiple peptides. Also, assays with a single 15-mer peptide, containing an identified epitope, did not always elicit a response providing validation that 15-mer peptides are not optimal antigens for detecting CD8+ T cells. Our findings further validate pooling-based epitope mapping strategies, which are critical for characterizing vaccine-induced T-cell responses and more broadly for informing iterative vaccine design. We also show ways to improve their application with computational peptide:MHC binding predictors that can accurately identify the optimal epitope within a 15-mer peptide and within a pool of 15-mer peptides.

Publication types

  • Clinical Trial, Phase II
  • Multicenter Study
  • Randomized Controlled Trial
  • Research Support, N.I.H., Extramural

MeSH terms

  • AIDS Vaccines / immunology*
  • AIDS Vaccines / therapeutic use
  • Adolescent
  • Adult
  • CD8-Positive T-Lymphocytes / cytology*
  • Clinical Trials as Topic
  • Cohort Studies
  • Double-Blind Method
  • Enzyme-Linked Immunospot Assay
  • Epitope Mapping / methods*
  • Epitopes, T-Lymphocyte / chemistry*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Peptides / chemistry*
  • Reproducibility of Results
  • Research Design
  • Sensitivity and Specificity
  • Young Adult

Substances

  • AIDS Vaccines
  • Epitopes, T-Lymphocyte
  • Peptides