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Increased peptide contacts govern high affinity binding of a modified TCR whilst maintaining a native pMHC docking mode

Journal Article · · Frontiers in Immunology
 [1]
  1. DOE/OSTI
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Natural T cell receptors (TCRs) generally bind to their cognate pMHC molecules with weak affinity and fast kinetics, limiting their use as therapeutic agents. Using phage display, we have engineered a high affinity version of the A6 wild-type TCR (A6wt), specific for the human leukocyte antigen (HLA-A*0201) complexed with human T cell lymphotropic virus type 111–19 peptide (A2-Tax). Mutations in just 4 residues in the CDR3β loop region of the A6wt TCR were selected that improved binding to A2-Tax by nearly 1000-fold. Biophysical measurements of this mutant TCR (A6c134) demonstrated that the enhanced binding was derived through favorable enthalpy and a slower off-rate. The structure of the free A6c134 TCR and the A6c134/A2-Tax complex revealed a native binding mode, similar to the A6wt/A2-Tax complex. However, concordant with the more favorable binding enthalpy, the A6c134TCR made increased contacts with the Tax peptide compared with the A6wt/A2- Tax complex, demonstrating a peptide-focused mechanism for the enhanced affinity that directly involved the mutated residues in the A6c134TCR CDR3β loop. This peptide-focused enhanced TCR binding may represent an important approach for developing antigen specific high affinity TCR reagents for use in T cell based therapies.
Research Organization:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1628044
Journal Information:
Frontiers in Immunology, Journal Name: Frontiers in Immunology Vol. 4; ISSN 1664-3224
Publisher:
Frontiers Research FoundationCopyright Statement
Country of Publication:
United States
Language:
English

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Cited By (9)

Changing the peptide specificity of a human T-cell receptor by directed evolution journal November 2014
Bench, Bedside, Toolbox: T-Cells Deliver on Every Level journal January 2014
An old Twist in HLA-A: CDR3α Hook up at an R65-joint journal May 2015
Using Global Analysis to Extend the Accuracy and Precision of Binding Measurements with T cell Receptors and Their Peptide/MHC Ligands journal January 2017
Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens journal February 2017
Stabilization of soluble high‐affinity T‐cell receptor with de novo disulfide bonds journal October 2019
The 3A6‐TCR/superagonist/HLA‐DR2a complex shows similar interface and reduced flexibility compared to the complex with self‐peptide journal July 2019
Human cytomegalovirus-specific T-cell receptor engineered for high affinity and soluble expression using mammalian cell display journal February 2019
The 3A6-TCR/superagonist/HLA-DR2a complex shows similar interface and reduced flexibility compared to the complex with self-peptide text January 2020

Figures / Tables (7)

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59 BASIC BIOLOGICAL SCIENCES
A6 TCR
Immunology
T cell
T cell receptor
crystal structure
high affinity TCR
human T leukocyte virus type 1
peptide-major histocompatibility complex
surface plasmon resonance