1. Affinity-matured HLA class II dimers for robust staining of antigen-specific CD4+ T cells.
- Author
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Sugata, Kenji, Matsunaga, Yukiko, Yamashita, Yuki, Nakatsugawa, Munehide, Guo, Tingxi, Halabelian, Levon, Ohashi, Yota, Saso, Kayoko, Rahman, Muhammed A., Anczurowski, Mark, Wang, Chung-Hsi, Murata, Kenji, Saijo, Hiroshi, Kagoya, Yuki, Ly, Dalam, Burt, Brian D., Butler, Marcus O., Mak, Tak W., and Hirano, Naoto
- Abstract
Peptide–major histocompatibility complex (pMHC) multimers enable the detection of antigen-specific T cells in studies ranging from vaccine efficacy to cancer immunotherapy. However, this technology is unreliable when applied to pMHC class II for the detection of CD4+ T cells. Here, using a combination of molecular biological and immunological techniques, we cloned sequences encoding human leukocyte antigen (HLA)-DP, HLA-DQ and HLA-DR molecules with enhanced CD4 binding affinity (with a Kd of 8.9 ± 1.1 µM between CD4 and affinity-matured HLA-DP4) and produced affinity-matured class II dimers that stain antigen-specific T cells better than conventional multimers in both in vitro and ex vivo analyses. Using a comprehensive library of dimers for HLA-DP4, which is the most frequent HLA allele in many ancestry groups, we mapped 103 HLA-DP4-restricted epitopes derived from diverse tumor-associated antigens and cloned the cognate T-cell antigen receptor (TCR) genes from in vitro-stimulated CD4+ T cells. The availability of affinity-matured class II dimers across HLA-DP, HLA-DQ and HLA-DR alleles will aid in the investigation of human CD4+ T-cell responses. CD4+ T cells are readily detected with class II MHC dimers. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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