Epitopes described in "ORFeome approach to the clonal, HLA allele-specific CD4 T-cell response to a complex pathogen in humans."

Article Authors:Lichen Jing; Stella Mayo McCaughey; D Huw Davies; Tiana M Chong; Phillip L Felgner; Stephen C De Rosa; Christopher B Wilson; David M Koelle
Article Title:ORFeome approach to the clonal, HLA allele-specific CD4 T-cell response to a complex pathogen in humans.
Reference Detail
Reference ID:1016588
Abstract:The CD4 T-cell response to vaccinia promotes antibody and long-term CD8 responses. HLA class II molecules present microbial epitopes to CD4 T-cells. In humans, at least 3 loci encode cell-surface peptide-binding HLA class II heterodimers. Using intracellular cytokine cytometry (ICC) assays, we determined that HLA DR had the strongest contribution to vaccinia antigen presentation. Among panels of vaccinia-restricted T-cell clones, most were DR-restricted but rare DQ-restricted clones were also recovered. Vaccinia has over 200 open reading frames (ORFs), providing a significant bottleneck to assigning fine specificity. To overcome this, we expressed each predicted vaccinia ORF using in vitro transcription and translation. Array-based pool proteins were used to rapidly assign fine specificity to each DQ-restricted clone and to a sample of HLA DR-restricted clones. Reactivity was confirmed using synthetic peptides for selected CD4 T-cell clones. This method should be broadly applicable to the study of large-genome, sequenced pathogens, and could also be used to investigate T-cell responses to cDNAs expressed in neoplastic and autoimmune disorders in which CD4 responses might be adaptive or harmful.
Affiliations:Department of Medicine, University of Washington, 1616 Eastlake Avenue East, Suite 251, Seattle, WA 98102, USA. lchjing@u.washington.edu.
Reference Type:Literature
PubMed ID:19520082
Journal:J Immunol Methods
Journal Volume:347
Article Pages:36-45
Journal ISSN:0022-1759
Article Chemical List:Antigens, Viral;Cytokines;DryVax vaccine;HLA-DQ Antigens;HLA-DR Antigens;Smallpox Vaccine
Article MeSH List:Animals; Antigen Presentation; Antigens, Viral(genetics; immunology); CD4-Positive T-Lymphocytes(immunology; virology); COS Cells; Cercopithecus aethiops; Cytokines(metabolism); Genome, Viral; HLA-DQ Antigens(immunology); HLA-DR Antigens(immunology); Humans; Hybridomas; Mice; Open Reading Frames; Proteomics(methods); Smallpox Vaccine(immunology); Vaccinia virus(genetics; immunology; pathogenicity)
Curation Last Updated:2015-06-05 01:46:57