Journal
IMMUNOGENETICS
Volume 65, Issue 5, Pages 357-370Publisher
SPRINGER
DOI: 10.1007/s00251-013-0684-y
Keywords
HLA Class II; Restriction; Transfectants; Epitopes; Population coverage; Polymorphism
Categories
Funding
- National Institutes of Health [N01-AI-900044C, AI-900048C, AI-100275]
- Bill and Melinda Gates Foundation [OPP1021972-3]
- Bill and Melinda Gates Foundation [OPP1021972] Funding Source: Bill and Melinda Gates Foundation
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Classic ways to determine MHC restriction involve inhibition with locus-specific antibodies and antigen presentation assays with panels of cell lines matched or mismatched at the various loci of interest. However, these determinations are often complicated by T cell epitope degeneracy and promiscuity. We describe a selection of 46 HLA DR, DQ, and DP specificities that provide worldwide population (phenotypic) coverage of almost 90 % at each locus, and account for over 66 % of all genes at each locus. This panel afforded coverage of at least four HLA class II alleles in over 95 % of the individuals in four study populations of diverse ethnicity from the USA and South Africa. Next, a panel of single HLA class II-transfected cell lines, corresponding to these 46 allelic variants was assembled, consisting of lines previously developed and 15 novel lines generated for the present study. The novel lines were validated by assessing their HLA class II expression by FACS analysis, the in vitro peptide binding activity of HLA molecules purified from the cell lines, and their antigen presenting capacity to T cell lines of known restriction. We also show that these HLA class II-transfected cell lines can be used to rapidly and unambiguously determine HLA restriction of epitopes recognized by an individual donor in a single experiment. This panel of lines will enable high throughput determination of HLA restriction, enabling better characterization of HLA class II-restricted T cell responses and facilitating the development of HLA tetrameric staining reagents.
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