Chang, Cynthia X L2; Tan, Anthony T2; Or, Ming Yan2; Toh, Kai Yee2; Lim, Pei Yiing2; Chia, Adeline S E2; Froesig, Thomas M2; Nadua, Karen D2; Oh, Hsueh-Ling J2; Leong, Hoe Nam2; Hadrup, Sine R1; Gehring, Adam J2; Tan, Yee-Joo2; Bertoletti, Antonio2; Grotenbreg, Gijsbert M2
1 Department of Haematology, Herlev and Gentofte Hospital, The Capital Region of Denmark2 unknown
Conditional ligands have enabled the high-throughput production of human leukocyte antigen (HLA) libraries that present defined peptides. Immunomonitoring platforms typically concentrate on restriction elements associated with European ancestry, and such tools are scarce for Asian HLA variants. We report 30 novel irradiation-sensitive ligands, specifically targeting South East Asian populations, which provide 93, 63, and 79% coverage for HLA-A, -B, and -C, respectively. Unique ligands for all 16 HLA types were constructed to provide the desired soluble HLA product in sufficient yield. Peptide exchange was accomplished for all variants as demonstrated by an ELISA-based MHC stability assay. HLA tetramers with redirected specificity could detect antigen-specific CD8(+) T-cell responses against human cytomegalovirus, hepatitis B (HBV), dengue virus (DENV), and Epstein-Barr virus (EBV) infections. The potential of this population-centric HLA library was demonstrated with the characterization of seven novel T-cell epitopes from severe acute respiratory syndrome coronavirus, HBV, and DENV. Posthoc analysis revealed that the majority of responses would be more readily identified by our unbiased discovery approach than through the application of state-of-the-art epitope prediction. This flow cytometry-based technology therefore holds considerable promise for monitoring clinically relevant antigen-specific T-cell responses in populations of distinct ethnicity.
European Journal of Immunology, 2013, Vol 43, Issue 4, p. 1109-20
Journal Article; Research Support, Non-U.S. Gov't; Amino Acid Sequence; Asian Continental Ancestry Group; CD8-Positive T-Lymphocytes; Epitopes, T-Lymphocyte; HLA Antigens; Humans; Ligands; Molecular Sequence Data; Peptides; Protein Multimerization; Protein Stability; Virus Diseases