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Journal: Frontiers in Immunology
Article Title: Identification of immunodominant T cell epitopes induced by natural Zika virus infection
doi: 10.3389/fimmu.2023.1247876
Figure Lengend Snippet: Immunoinformatic strategy to identify MHC I and MHC II T cell epitopes. (A) The entire sequence of ZIKV strain PRVABC59 and 104 additional ZIKV strains were parsed into 9- and 10-mer frames, and conserved sequences predicted to bind to 6 MHC I supertypes and 9 common MHC II alleles were identified using Conservatrix and EpiMatrix algorithms. Promiscuous panDR clusters (DR) were generated from overlapping frames predicted to bind multiple MHC II alleles using the ClustiMer algorithm (16-27 amino acids). Sequences with high homology with human proteins (potential human cross-reactive sequences) were excluded using the JanusMatrix algorithm. (B) Shown is an example (Cluster DR1) of an EpiMatrix Cluster Report. Z-score indicates the potential of a 9-mer frame to bind to a given HLA allele; the strength of the score is indicated by the blue shading. Scores in the top 5% (Z-score ≥1.64) are considered “Hits”. *Z-scores in the top 10% are considered elevated, other scores grayed for simplicity. Frames containing four or more alleles scoring above 1.64 are referred to as EpiBars and are highlighted in yellow. Flanking amino acids, added to stabilize the cluster during in vitro testing, are underlined. The EpiMatrix Cluster Score is derived from the sum of Z-scores of EpiMatrix hits, normalized for the length of the cluster. Thus, EpiMatrix Cluster Score represents the excess or shortfall in predicted aggregate immunogenicity relative to a random peptide standard. Cluster Scores above 10 indicate significant potential for promiscuous response. (C) The 25-30 conserved ZIKV peptides predicted to bind the 6 MHC I supertypes and 33 clusters predicted to promiscuously bind human HLA-DR were synthesized for further study. The locations of each putative epitope or cluster within the ZIKV polyprotein are shown.
Article Snippet:
Techniques: Sequencing, Generated, In Vitro, Derivative Assay, Immunopeptidomics, Synthesized