MHC class II tetramers engineered for enhanced binding to CD4 improve detection of antigen-specific T cells
- 3 May 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Biotechnology
- Vol. 39 (8), 943-948
- https://doi.org/10.1038/s41587-021-00893-9
Abstract
The ability to identify T cells that recognize specific peptide antigens bound to major histocompatibility complex (MHC) molecules has enabled enumeration and molecular characterization of the lymphocytes responsible for cell-mediated immunity. Fluorophore-labeled peptide:MHC class I (p:MHCI) tetramers are well-established reagents for identifying antigen-specific CD8+ T cells by flow cytometry, but efforts to extend the approach to CD4+ T cells have been less successful, perhaps owing to lower binding strength between CD4 and MHC class II (MHCII) molecules. Here we show that p:MHCII tetramers engineered by directed evolution for enhanced CD4 binding outperform conventional tetramers for the detection of cognate T cells. Using the engineered tetramers, we identified about twice as many antigen-specific CD4+ T cells in mice immunized against multiple peptides than when using traditional tetramers. CD4 affinity-enhanced p:MHCII tetramers, therefore, allow direct sampling of antigen-specific CD4+ T cells that cannot be accessed with conventional p:MHCII tetramer technology. These new reagents could provide a deeper understanding of the T cell repertoire.Keywords
Funding Information
- U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (T32 AI083196, T32 AI007313, R01 AI096879, R01 AI143826, R01 AI039614)
This publication has 32 references indexed in Scilit:
- Affinity maturation of human CD4 by yeast surface display and crystal structure of a CD4–HLA-DR1 complexProceedings of the National Academy of Sciences of the United States of America, 2011
- Quantitative impact of thymic selection on Foxp3 + and Foxp3 − subsets of self-peptide/MHC class II-specific CD4 + T cellsProceedings of the National Academy of Sciences of the United States of America, 2011
- The Tetramer TransformationThe Journal of Immunology, 2011
- Fast on-rates allow short dwell time ligands to activate T cellsProceedings of the National Academy of Sciences of the United States of America, 2010
- Naive CD4+ T Cell Frequency Varies for Different Epitopes and Predicts Repertoire Diversity and Response MagnitudeImmunity, 2007
- Overview of Gene Targeting by Homologous RecombinationCurrent Protocols in Molecular Biology, 2006
- T Cell Receptor Binding to a pMHCII Ligand Is Kinetically Distinct from and Independent of CD4Online Journal of Public Health Informatics, 2001
- T Cell Receptor and Coreceptor CD8αα Bind Peptide-MHC Independently and with Distinct KineticsImmunity, 1999
- Phenotypic Analysis of Antigen-Specific T LymphocytesScience, 1996
- Presentation of endogenous viral proteins in association with major histocompatibility complex class II: On the role of intracellular compartmentalization, invariant chain and the TAP transporter systemEuropean Journal of Immunology, 1995