Engineering and characterizing human T cell receptors for cancer immunotherapies
type: CreativeWork
description: The T cell receptor (TCR) is an heterodimer that binds to a short peptide bound to a product of the major histocompatibility complex (MHC). The chains each contain variable (V) and constant (C) region domains, followed by a transmembrane region. Each V domain contains three loops, called complementarity-determining regions (CDR1, CDR2, and CDR3), which interact with the peptide (pep)MHC antigen. The conserved docking angle of the six CDRs over the pepMHC antigens is thought to confer maximal signaling capabilities and it places the two most hypervariable loops (CDR3s) over the most diverse portion of the antigen (the peptide). Recent adoptive T cell trials have shown promising results using TCRs directed towards pepMHC complexes expressed in various types of malignancies. The process of immunological tolerance leads to the deletion of T cells with TCRs against possible cancer peptide self- antigens. Those TCRs that remain typically do not have the required affinity to mediate a sufficient T cell response. To overcome this, TCRs can be engineered for higher- affinity ex vivo and reintroduced into patient’s peripheral T cells. In chapter two, a TCR with a switch in peptide specificity was characterized. In previous studies, a TCR called RD1-MART1HIGH was engineered to bind to the cancer peptide MART1/HLA-A2, using libraries of mutants form the parental TCR, which is specific for a viral peptide, Tax/HLA-A2. The switch in peptide specificity was achieved by a limited number of CDR substitutions. To understand which residues contributed to the new peptide specificity and TCR binding, single codon libraries were created at every CDR residue. Libraries were pooled and sorted for binding to MART1/HLA-A2 to produce a sequence fitness landscape, which measured the effect that each individual substitution had on pepMHC binding. The sequence fitness landscape of RD1- MART1HIGH was compared to a sequence fitness landscape of its parent TCR A6 to identify the key residues involv…
abstract: The T cell receptor (TCR) is an heterodimer that binds to a short peptide bound to a product of the major histocompatibility complex (MHC). The chains each contain variable (V) and constant (C) region domains, followed by a transmembrane region. Each V domain contains three loops, called complementarity-determining regions (CDR1, CDR2, and CDR3), which interact with the peptide (pep)MHC antigen. The conserved docking angle of the six CDRs over the pepMHC antigens is thought to confer maximal signaling capabilities and it places the two most hypervariable loops (CDR3s) over the most diverse portion of the antigen (the peptide). Recent adoptive T cell trials have shown promising results using TCRs directed towards pepMHC complexes expressed in various types of malignancies. The process of immunological tolerance leads to the deletion of T cells with TCRs against possible cancer peptide self- antigens. Those TCRs that remain typically do not have the required affinity to mediate a sufficient T cell response. To overcome this, TCRs can be engineered for higher- affinity ex vivo and reintroduced into patient’s peripheral T cells. In chapter two, a TCR with a switch in peptide specificity was characterized. In previous studies, a TCR called RD1-MART1HIGH was engineered to bind to the cancer peptide MART1/HLA-A2, using libraries of mutants form the parental TCR, which is specific for a viral peptide, Tax/HLA-A2. The switch in peptide specificity was achieved by a limited number of CDR substitutions. To understand which residues contributed to the new peptide specificity and TCR binding, single codon libraries were created at every CDR residue. Libraries were pooled and sorted for binding to MART1/HLA-A2 to produce a sequence fitness landscape, which measured the effect that each individual substitution had on pepMHC binding. The sequence fitness landscape of RD1- MART1HIGH was compared to a sequence fitness landscape of its parent TCR A6 to identify the key residues involv…
author: Harris, Daniel Thomas
language: en
keywords: Immunotherapy, T Cell Receptor
date published: 2017-03-01T17:00:44Z
date modified: 2026-07-22T22:26:37Z
provider: University of Illinois - Urbana-Champaign
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Illinois at Urbana-Champaign

PropertyValue
property ID: Handle
value: http://hdl.handle.net/2142/95531