Affinity-matured HLA class II dimers for robust staining of antigen-specific CD4+ T cells

Kenji Sugata; Yukiko Matsunaga; Yuki Yamashita; Munehide Nakatsugawa; Tingxi Guo; Levon Halabelian; Yota Ohashi; Kayoko Saso; Muhammed A. Rahman; Mark Anczurowski; Chung Hsi Wang; Kenji Murata; Hiroshi Saijo; Yuki Kagoya; Dalam Ly; Brian D. Burt; Marcus O. Butler; Tak W. Mak; Naoto Hirano

doi:10.1038/s41587-021-00836-4

Affinity-matured HLA class II dimers for robust staining of antigen-specific CD4⁺ T cells

Kenji Sugata, Yukiko Matsunaga, Yuki Yamashita, Munehide Nakatsugawa, Tingxi Guo, Levon Halabelian, Yota Ohashi, Kayoko Saso, Muhammed A. Rahman, Mark Anczurowski, Chung Hsi Wang, Kenji Murata, Hiroshi Saijo, Yuki Kagoya, Dalam Ly, Brian D. Burt, Marcus O. Butler, Tak W. Mak, Naoto Hirano

Research output: Contribution to journal › Article › peer-review

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 K_d 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.

Original language	English
Pages (from-to)	958-967
Number of pages	10
Journal	Nature Biotechnology
Volume	39
Issue number	8
DOIs	https://doi.org/10.1038/s41587-021-00836-4
Publication status	Published - 2021 Aug
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology
Molecular Medicine
Biomedical Engineering

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1038/s41587-021-00836-4

Cite this

Sugata, K., Matsunaga, Y., Yamashita, Y., Nakatsugawa, M., Guo, T., Halabelian, L., Ohashi, Y., Saso, K., Rahman, M. A., Anczurowski, M., Wang, C. H., Murata, K., Saijo, H., Kagoya, Y., Ly, D., Burt, B. D., Butler, M. O., Mak, T. W., & Hirano, N. (2021). Affinity-matured HLA class II dimers for robust staining of antigen-specific CD4⁺ T cells. Nature Biotechnology, 39(8), 958-967. https://doi.org/10.1038/s41587-021-00836-4

Sugata, K, Matsunaga, Y, Yamashita, Y, Nakatsugawa, M, Guo, T, Halabelian, L, Ohashi, Y, Saso, K, Rahman, MA, Anczurowski, M, Wang, CH, Murata, K, Saijo, H, Kagoya, Y, Ly, D, Burt, BD, Butler, MO, Mak, TW & Hirano, N 2021, 'Affinity-matured HLA class II dimers for robust staining of antigen-specific CD4⁺ T cells', Nature Biotechnology, vol. 39, no. 8, pp. 958-967. https://doi.org/10.1038/s41587-021-00836-4

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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.",

author = "Kenji Sugata and Yukiko Matsunaga and Yuki Yamashita and Munehide Nakatsugawa and Tingxi Guo and Levon Halabelian and Yota Ohashi and Kayoko Saso and Rahman, {Muhammed A.} and Mark Anczurowski and Wang, {Chung Hsi} and Kenji Murata and Hiroshi Saijo and Yuki Kagoya and Dalam Ly and Burt, {Brian D.} and Butler, {Marcus O.} and Mak, {Tak W.} and Naoto Hirano",

note = "Funding Information: This work was supported by the Ontario Institute for Cancer Research Clinical Investigator Award IA-039 (N.H.), the Princess Margaret Cancer Centre Innovation Accelerator Fund (N.H.), the Ira Schneider Memorial Cancer Research Foundation (N.H.), the Princess Margaret Cancer Foundation (N.H. and M.O.B.), the Uehara Memorial Foundation Research Fellowship Program (K. Sugata), the Mitacs Internship (K.M.), the Japan Society for the Promotion of Science Postdoctoral Fellowship for Overseas Researchers and the Guglietti fellowship (Y.K.), the Province of Ontario (T.G. and M.A.), the Natural Sciences and Engineering Research Council of Canada Postgraduate Scholarship (T.G.) and the Frederick Banting and Charles Best Canada Graduate Scholarship (C.-H.W.). Publisher Copyright: {\textcopyright} 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.",

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T1 - Affinity-matured HLA class II dimers for robust staining of antigen-specific CD4+ T cells

AU - Sugata, Kenji

AU - Matsunaga, Yukiko

AU - Yamashita, Yuki

AU - Nakatsugawa, Munehide

AU - Guo, Tingxi

AU - Halabelian, Levon

AU - Ohashi, Yota

AU - Saso, Kayoko

AU - Rahman, Muhammed A.

AU - Anczurowski, Mark

AU - Wang, Chung Hsi

AU - Murata, Kenji

AU - Saijo, Hiroshi

AU - Kagoya, Yuki

AU - Ly, Dalam

AU - Burt, Brian D.

AU - Butler, Marcus O.

AU - Mak, Tak W.

AU - Hirano, Naoto

N1 - Funding Information: This work was supported by the Ontario Institute for Cancer Research Clinical Investigator Award IA-039 (N.H.), the Princess Margaret Cancer Centre Innovation Accelerator Fund (N.H.), the Ira Schneider Memorial Cancer Research Foundation (N.H.), the Princess Margaret Cancer Foundation (N.H. and M.O.B.), the Uehara Memorial Foundation Research Fellowship Program (K. Sugata), the Mitacs Internship (K.M.), the Japan Society for the Promotion of Science Postdoctoral Fellowship for Overseas Researchers and the Guglietti fellowship (Y.K.), the Province of Ontario (T.G. and M.A.), the Natural Sciences and Engineering Research Council of Canada Postgraduate Scholarship (T.G.) and the Frederick Banting and Charles Best Canada Graduate Scholarship (C.-H.W.). Publisher Copyright: © 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.

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