title: Sequence-based in silico analysis of well studied hepatitis C virus epitopes and their variants in other genotypes (particularly genotype 5a) against South African human leukocyte antigen backgrounds.
abstract: BACKGROUND
  Host genetics influence the outcome of HCV disease. HCV is also highly mutable and escapes host immunity. HCV genotypes are geographically distributed and HCV subtypes have been shown to have distinct repertoires of HLA-restricted viral epitopes which explains the lack of cross protection across genotypes observed in some studies. Despite this, immune databases and putative epitope vaccines concentrate almost exclusively on HCV genotype 1 class I-epitopes restricted by the HLA-A*02 allele. While both genotype and allele predominate in developed countries, we hypothesise that HCV variation and population genetics will affect the efficacy of proposed epitope vaccines in South Africa. This in silico study investigates HCV viral variability within well-studied epitopes identified in genotype 1 and uses algorithms to predict the immunogenicity of their variants from other less studied genotypes and thus rate the most promising vaccine candidates for the South African population. Six class I- and seven class II- restricted epitope sequences within the core, NS3, NS4B and NS5B regions were compared across the six HCV genotypes using local genotype 5a sequence data together with global data. Common HLA alleles in the South African population are A30:01, A02:01, B58:02, B07:02; DRB1*13:01 and DRB1*03:01. Epitope binding to 13 class I- and 8 class -II alleles were described using web-based prediction servers, Immune Epitope Database, (IEDB) and Propred. Online population coverage tools were used to assess vaccine efficacy.
  
  
  RESULTS
  Despite the homogeneity of genotype 1 and genotype 5 over the epitopes, there was limited promiscuity to local HLA-alleles.Host differences will make a putative vaccine less effective in South Africa. Of the 6 well-characterized class I- epitopes, only 2 class I- epitopes were promiscuous and 3 of the 7 class-II epitopes were better conserved and promiscuous. By fine tuning the putative vaccine using an optimal cocktail of genotype 1 and 5a epitopes and local HLA data, the coverage was raised from 65.85% to 91.87% in South African Blacks.
  
  
  CONCLUSIONS
  While in vivo and in vitro studies are needed to confirm immunogenic epitopes, in silico HCV epitope vaccine design which takes into account HCV variation and host allele frequency will maximize population coverage in different ethnic groups.
authors.affiliation: Specialized Molecular Diagnostics, Hepatitis Unit, National Institute for Communicable Diseases, National Health Laboratory Services, Johannesburg, South Africa. niship@nicd.ac.za
authors.forename: Nishi
authors.initials: N
authors.lastname: Prabdial-Sing
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Puren
authors.forename: Sheila M
authors.initials: SM
authors.lastname: Bowyer
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D019295:Computational Biology / Q000379:methods*; D017124:Conserved Sequence; D030562:Databases, Protein; D005838:Genotype; D006680:HLA Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D020539:Sequence Analysis, Protein; D013019:South Africa
chemical_list: D006680:HLA Antigens; D016056:Immunodominant Epitopes