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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
A strategy to determine HLA class II restriction broadly covering the DR, DP, and DQ allelic variants most commonly expressed in the general population
Immunogenetics, Volume 65, No. 5, Year 2013
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Description
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. © 2013 Springer-Verlag Berlin Heidelberg.
Authors & Co-Authors
McKinney, Denise M.
United States, La Jolla
La Jolla Institute for Immunology
Southwood, Scott
United States, La Jolla
La Jolla Institute for Immunology
Hinz, Denise
United States, La Jolla
La Jolla Institute for Immunology
Oseroff, Carla
United States, La Jolla
La Jolla Institute for Immunology
Lindestam Arlehamn, Cecilia S.
United States, La Jolla
La Jolla Institute for Immunology
Schulten, Véronique
United States, La Jolla
La Jolla Institute for Immunology
Taplitz, Randy Allison
United States, La Jolla
University of California, San Diego
Broide, David H.
United States, La Jolla
University of California, San Diego
Hanekom, Willem Albert
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Scriba, Thomas J.
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Wood, Robert A.
United States, Baltimore
Johns Hopkins University
Alam, Rapheul
United States, Denver
National Jewish Health
Peters, Bjoern S.
United States, La Jolla
La Jolla Institute for Immunology
Sidney, John
United States, La Jolla
La Jolla Institute for Immunology
Sette, Alessandro D.
United States, La Jolla
La Jolla Institute for Immunology
Statistics
Citations: 73
Authors: 15
Affiliations: 5
Identifiers
Doi:
10.1007/s00251-013-0684-y
ISSN:
00937711
e-ISSN:
14321211
Research Areas
Health System And Policy
Study Design
Cross Sectional Study
Study Approach
Quantitative
Study Locations
South Africa