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39 items found (0 items not approved), displaying 1 to 10.[First/Prev] 1, 2, 3, 4 [Next/Last]

Instruments / Technical Area	Publication Details	Published
I03-Macromolecular Crystallography I04-Macromolecular Crystallography	HLA A24:02–restricted T cell receptors cross-recognize bacterial and preproinsulin peptides in type 1 diabetes Garry Dolton , Anna Bulek , Aaron Wall , Hannah Thomas , Jade R. Hopkins , Cristina Rius , Sarah A. E. Galloway , Thomas Whalley , Li Rong Tan , Théo Morin , Nader Omidvar , Anna Fuller , Katie Topley , Md Samiul Hasan , Shikha Jain , Nirupa D’souza , Thomas Hodges-Hoyland , Owen B. Spiller , Deborah Kronenberg-Versteeg , Barbara Szomolay , Hugo A. Van Den Berg , Lucy C. Jones , Mark Peakman , David K. Cole , Pierre J. Rizkallah , Andrew K. Sewell Journal Of Clinical Investigation 134* DOI: 10.1172/JCI164535 Diamond Proposal Number(s): [10462, 18812] Open Access Show Abstract ▼ Abstract: CD8+ T cells destroy insulin-producing pancreatic β cells in type 1 diabetes through HLA class I–restricted presentation of self-antigens. Combinatorial peptide library screening was used to produce a preferred peptide recognition landscape for a patient-derived T cell receptor (TCR) that recognized the preproinsulin-derived (PPI-derived) peptide sequence LWMRLLPLL in the context of disease risk allele HLA A24:02. Data were used to generate a strong superagonist peptide, enabling production of an autoimmune HLA A24:02–peptide–TCR structure by crystal seeding. TCR binding to the PPI epitope was strongly focused on peptide residues Arg4 and Leu5, with more flexibility at other positions, allowing the TCR to strongly engage many peptides derived from pathogenic bacteria. We confirmed an epitope from Klebsiella that was recognized by PPI-reactive T cells from 3 of 3 HLA A24:02+ patients. Remarkably, the same epitope selected T cells from 7 of 8 HLA A24+ healthy donors that cross-reacted with PPI, leading to recognition and killing of HLA A*24:02+ cells expressing PPI. These data provide a mechanism by which molecular mimicry between pathogen and self-antigens could have resulted in the breaking of self-tolerance to initiate disease.	Sep 2024
I03-Macromolecular Crystallography	A targeted single mutation in influenza A virus universal epitope transforms immunogenicity and protective immunity via CD4+ T cell activation Sarah Hulin-Curtis , James K. Geary , Bruce J. Maclachlan , Danny M. Altmann , Laury Baillon , David K. Cole , Alexander Greenshields-Watson , Sophie J. Hesketh , Ian R. Humphreys , Ian M. Jones , Sarah N. Lauder , Georgina H. Mason , Kathryn Smart , D. Oliver Scourfield , Jake Scott , Ksenia Sukhova , Richard J. Stanton , Aaron Wall , Pierre J. Rizkallah , Wendy S. Barclay , Awen Gallimore , Andrew Godkin Cell Reports 43 DOI: 10.1016/j.celrep.2024.114259 Diamond Proposal Number(s): [10462] Open Access Show Abstract ▼ Abstract: CD4+ T cells are central to adaptive immunity. Their role in cross-protection in viral infections such as influenza and severe acute respiratory syndrome (SARS) is well documented; however, molecular rules governing T cell receptor (TCR) engagement of peptide-human leukocyte antigen (pHLA) class II are less understood. Here, we exploit an aspect of HLA class II presentation, the peptide-flanking residues (PFRs), to “tune” CD4+ T cell responses within an in vivo model system of influenza. Using a recombinant virus containing targeted substitutions at immunodominant HLA-DR1 epitopes, we demonstrate limited weight loss and improved clinical scores after heterosubtypic re-challenge. We observe enhanced protection linked to lung-derived influenza-specific CD4+ and CD8+ T cells prior to re-infection. Structural analysis of the ternary TCR:pHLA complex identifies that flanking amino acids influence side chains in the core 9-mer peptide, increasing TCR affinity. Augmentation of CD4+ T cell immunity is achievable with a single mutation, representing a strategy to enhance adaptive immunity that is decoupled from vaccine modality.	Jun 2024
I03-Macromolecular Crystallography I04-Macromolecular Crystallography	Promiscuous recognition of MR1 drives self-reactive mucosal-associated invariant T cell responses Andrew Chancellor , Robert A. Simmons , Rahul C. Khanolkar , Vladimir Nosi , Aisha Beshirova , Giuliano Berloffa , Rodrigo Colombo , Vijaykumar Karuppiah , Johanne M. Pentier , Vanessa Tubb , Hemza Ghadbane , Richard J. Suckling , Keith Page , Rory M. Crean , Alessandro Vacchini , Corinne De Gregorio , Verena Schaefer , Daniel Constantin , Thomas Gligoris , Angharad Lloyd , Miriam Hock , Velupillai Srikannathasan , Ross A. Robinson , Gurdyal S. Besra , Marc W. Van Der Kamp , Lucia Mori , Raffaele Calogero , David K. Cole , Gennaro De Libero , Marco Lepore Journal Of Experimental Medicine 220 DOI: 10.1084/jem.20221939 Diamond Proposal Number(s): [22870, 28224] Show Abstract ▼ Abstract: Mucosal-associated invariant T (MAIT) cells use canonical semi-invariant T cell receptors (TCR) to recognize microbial riboflavin precursors displayed by the antigen-presenting molecule MR1. The extent of MAIT TCR crossreactivity toward physiological, microbially unrelated antigens remains underexplored. We describe MAIT TCRs endowed with MR1-dependent reactivity to tumor and healthy cells in the absence of microbial metabolites. MAIT cells bearing TCRs crossreactive toward self are rare but commonly found within healthy donors and display T-helper-like functions in vitro. Experiments with MR1-tetramers loaded with distinct ligands revealed significant crossreactivity among MAIT TCRs both ex vivo and upon in vitro expansion. A canonical MAIT TCR was selected on the basis of extremely promiscuous MR1 recognition. Structural and molecular dynamic analyses associated promiscuity to unique TCRβ-chain features that were enriched within self-reactive MAIT cells of healthy individuals. Thus, self-reactive recognition of MR1 represents a functionally relevant indication of MAIT TCR crossreactivity, suggesting a potentially broader role of MAIT cells in immune homeostasis and diseases, beyond microbial immunosurveillance.	Sep 2023
I03-Macromolecular Crystallography I04-1-Macromolecular Crystallography (fixed wavelength) I04-Macromolecular Crystallography	Targeting of multiple tumor-associated antigens by individual T cell receptors during successful cancer immunotherapy Garry Dolton , Cristina Rius , Aaron Wall , Barbara Szomolay , Valentina Bianchi , Sarah A. E. Galloway , Md Samiul Hasan , Théo Morin , Marine E. Caillaud , Hannah L. Thomas , Sarah Theaker , Li Rong Tan , Anna Fuller , Katie Topley , Mateusz Legut , Meriem Attaf , Jade R. Hopkins , Enas Behiry , Joanna Zabkiewicz , Caroline Alvares , Angharad Lloyd , Amber Rogers , Peter Henley , Christopher Fegan , Oliver Ottmann , Stephen Man , Michael D. Crowther , Marco Donia , Inge Marie Svane , David K. Cole , Paul E. Brown , Pierre Rizkallah , Andrew K. Sewell Cell 33 DOI: 10.1016/j.cell.2023.06.020 Open Access Show Abstract ▼ Abstract: The T cells of the immune system can target tumors and clear solid cancers following tumor-infiltrating lymphocyte (TIL) therapy. We used combinatorial peptide libraries and a proteomic database to reveal the antigen specificities of persistent cancer-specific T cell receptors (TCRs) following successful TIL therapy for stage IV malignant melanoma. Remarkably, individual TCRs could target multiple different tumor types via the HLA A∗02:01-restricted epitopes EAAGIGILTV, LLLGIGILVL, and NLSALGIFST from Melan A, BST2, and IMP2, respectively. Atomic structures of a TCR bound to all three antigens revealed the importance of the shared x-x-x-A/G-I/L-G-I-x-x-x recognition motif. Multi-epitope targeting allows individual T cells to attack cancer in several ways simultaneously. Such “multipronged” T cells exhibited superior recognition of cancer cells compared with conventional T cell recognition of individual epitopes, making them attractive candidates for the development of future immunotherapies.	Jul 2023
I04-1-Macromolecular Crystallography (fixed wavelength)	Development of a low seroprevalence, αvβ6 integrin selective virotherapy based on human adenovirus type 10 Emily A. Bates , James A. Davies , Jana Váňová , Davor Nestić , Valerie S. Meniel , Sarah Koushyar , Tabitha G. Cunliffe , Rosie M. Mundy , Elise Moses , Hanni K. Uusi-Kerttula , Alexander T. Baker , David K. Cole , Dragomira Majhen , Pierre J. Rizkallah , Toby Phesse , John D. Chester , Alan L. Parker Molecular Therapy - Oncolytics DOI: 10.1016/j.omto.2022.03.007 Diamond Proposal Number(s): [18812] Open Access Show Abstract ▼ Abstract: Oncolytic virotherapies (OV) hold immense clinical potential. OV based on human adenoviruses (HAdV) derived from HAdV with naturally low rates of pre-existing immunity will be beneficial for future clinical translation. We generated a low seroprevalence HAdV-D10 serotype vector incorporating an αvβ6 integrin selective peptide, A20, to target αvβ6 positive tumour cell types. HAdV-D10 has limited natural tropism. Structural and biological studies of HAdV-D10 knob protein highlighted low affinity engagement with native adenoviral receptors CAR and sialic acid. HAdV-D10 fails to engage blood coagulation Factor X, potentially eliminating “off-target” hepatic sequestration in vivo. We engineered A20 peptide that selectively binds αvβ6 integrin into the DG loop of HAdV-D10 fiber knob. Assays in αvβ6+ cancer cell lines, demonstrated significantly increased transduction mediated by αvβ6 targeted variants compared to controls, confirmed microscopically. HAdV-D10.A20 resisted neutralization by neutralizing HAdV-C5 sera. Systemic delivery of HAdV-D10.A20 resulted in significantly increased GFP expression in BT20 tumours. Replication competent HAdV-D10.A20 demonstrated αvβ6 integrin selective cell killing in vitro and in vivo. HAdV-D10 possesses characteristics of a promising virotherapy, combining low seroprevalence, weak receptor interactions and reduced off-target uptake. Incorporation of an αvβ6 integrin selective peptide resulted in HAdV-D10.A20, with significant potential for clinical translation.	Mar 2022
I03-Macromolecular Crystallography I04-1-Macromolecular Crystallography (fixed wavelength) I04-Macromolecular Crystallography	Structure‐guided stabilization of pathogen‐derived peptide‐HLA‐E complexes using non‐natural amino acids conserves native TCR recognition Claire Barber , Victoria Arena De Souza , Rachel L. Paterson , Magdalena Martin-Urdiroz , Nitha Charles Mulakkal , Velupillai Srikannathasan , Mary Connolly , Gwilym Phillips , Tein Foong-Leong , Robert Pengelly , Vijaykumar Karuppiah , Tressan Grant , Marcin Dembek , Anil Verma , Dawn Gibbs-Howe , Thomas H. Blicher , Andrew Knox , Ross A. Robinson , David K. Cole , Sarah Leonard European Journal Of Immunology DOI: 10.1002/eji.202149745 Open Access Show Abstract ▼ Abstract: The non-polymorphic class Ib molecule, human leukocyte antigen (HLA)-E, primarily presents peptides from HLA class Ia leader peptides, providing an inhibitory signal to NK cells via CD94/NKG2 interactions. Although peptides of pathogenic origin can also be presented by HLA-E to T cells, the molecular basis underpinning their role in antigen surveillance is largely unknown. Here, we solved a co-complex crystal structure of a T cell receptor (TCR) with an HLA-E presented peptide (pHLA-E) from bacterial (Mycobacterium tuberculosis) origin, and the first TCR-pHLA-E complex with a non-canonically presented peptide from viral (human immuno-deficiency virus; HIV) origin. The structures provided a molecular foundation to develop a novel method to introduce cysteine traps using non-natural amino acid chemistry that stabilized pHLA-E complexes whilst maintaining native interface contacts between the TCRs and different pHLA-E complexes. These pHLA-E monomers could be used to isolate pHLA-E specific T cells, with obvious utility for studying pHLA-E restricted T cells, and for the identification of putative therapeutic TCRs.	Feb 2022
I02-Macromolecular Crystallography I03-Macromolecular Crystallography	Synthetic peptides with inadvertent chemical modifications can activate potentially autoreactive T cells Stephen Man , James E. Redman , Deborah L. Cross , David K. Cole , Ilona Can , Bethan Davies , Shaikh Shimaz Hashimdeen , Reiss Reid , Sian Llewellyn-Lacey , Kelly L. Miners , Kristin Ladell , Anya Lissina , Paul E. Brown , Linda Wooldridge , David A. Price , Pierre J. Rizkallah The Journal Of Immunology DOI: 10.4049/jimmunol.2000756 Diamond Proposal Number(s): [10462, 14843] Show Abstract ▼ Abstract: The human CD8+ T cell clone 6C5 has previously been shown to recognize the tert-butyl-modified Bax161–170 peptide LLSY(3-tBu)FGTPT presented by HLA-A02:01. This nonnatural epitope was likely created as a by-product of fluorenylmethoxycarbonyl protecting group peptide synthesis and bound poorly to HLA-A02:01. In this study, we used a systematic approach to identify and characterize natural ligands for the 6C5 TCR. Functional analyses revealed that 6C5 T cells only recognized the LLSYFGTPT peptide when tBu was added to the tyrosine residue and did not recognize the LLSYFGTPT peptide modified with larger (di-tBu) or smaller chemical groups (Me). Combinatorial peptide library screening further showed that 6C5 T cells recognized a series of self-derived peptides with dissimilar amino acid sequences to LLSY(3-tBu)FGTPT. Structural studies of LLSY(3-tBu)FGTPT and two other activating nonamers (IIGWMWIPV and LLGWVFAQV) in complex with HLA-A*02:01 demonstrated similar overall peptide conformations and highlighted the importance of the position (P) 4 residue for T cell recognition, particularly the capacity of the bulky amino acid tryptophan to substitute for the tBu-modified tyrosine residue in conjunction with other changes at P5 and P6. Collectively, these results indicated that chemical modifications directly altered the immunogenicity of a synthetic peptide via molecular mimicry, leading to the inadvertent activation of a T cell clone with unexpected and potentially autoreactive specificities	Jul 2021
I03-Macromolecular Crystallography	The fiber knob protein of human adenovirus type 49 mediates highly efficient and promiscuous infection of cancer cell lines using a novel cell entry mechanism Alexander T. Baker , James A. Davies , Emily A. Bates , Elise Moses , Rosie M. Mundy , Gareth Marlow , David K. Cole , Carly M. Bliss , Pierre J. Rizkallah , Alan L. Parker Journal Of Virology DOI: 10.1128/JVI.01849-20 Diamond Proposal Number(s): [18812] Open Access Show Abstract ▼ Abstract: The human adenovirus (HAdV) phylogenetic tree is diverse, divided across seven species and comprising over 100 individual types. Species D HAdV are rarely isolated with low rates of pre-existing immunity, making them appealing for therapeutic applications. Several species D vectors have been developed as vaccines against infectious diseases where they induce robust immunity in pre-clinical models and early phase clinical trials. However, many aspects of the basic virology of species D HAdV, including their basic receptor usage and means of cell entry, remain understudied. Here, we investigated HAdV-D49, which previously has been studied for vaccine and vascular gene transfer applications. We generated a pseudotyped HAdV-C5 presenting the HAdV-D49 fiber knob protein (HAdV-C5/D49K). This pseudotyped vector was efficient at infecting cells devoid of all known HAdV receptors, indicating HAdV-D49 uses an unidentified cellular receptor. Conversely, a pseudotyped vector presenting the fiber knob protein of the closely related HAdV-D30 (HAdV-C5/D30K), differing in four amino acids to HAdV-D49, failed to demonstrate the same tropism. These four amino acid changes resulted in a change in isoelectric point of the knob protein, with HAdV-D49K possessing a basic apical region compared to a more acidic region in HAdV-D30K. Structurally and biologically we demonstrate that HAdV-D49 knob protein is unable to engage CD46, while potential interaction with CAR is extremely limited by extension of the DG loop. HAdV-C5/49K efficiently transduced cancer cell lines of pancreatic, breast, lung, oesophageal and ovarian origin, indicating it may have potential for oncolytic virotherapy applications, especially for difficult to transduce tumor types.	Dec 2020
I03-Macromolecular Crystallography	Identification of a superagonist variant of the immunodominant Yellow fever virus epitope NS4b 214-222 by combinatorial peptide library screening Amandine Bovay , Vincent Zoete , Pierre J. Rizkallah , Konrad Beck , Philippe Delbreil , Daniel E. Speiser , David K. Cole , Silvia A. Fuertes Marraco Molecular Immunology 125, 43 - 50 DOI: 10.1016/j.molimm.2020.06.025 Open Access Show Abstract ▼ Abstract: The CD8 T cell response to the HLA-A2-restricted epitope LLWNGPMAV (LLW) of the non-structural protein 4b of Yellow Fever Virus (YFV) is remarkably immunodominant, highly prevalent and powerful in YFV-vaccinated humans. Here we used a combinatorial peptide library screening in the context of an A2/LLW-specific CD8 T cell clone to identify a superagonist that features a methionine to isoleucine substitution at position 7. Based on in silico modeling, the functional enhancement of this LLW-7I mutation was associated with alterations in the structural dynamics of the peptide in the major histocompatibility complex (pMHC) binding with the T cell receptor (TCR). While the TCR off-rate of LLW-7I pMHC is comparable to the wild type peptide, the rigidity of the 7I peptide seems to confer less entropy loss upon TCR binding. This LLW-7I superagonist is an example of improved functionality in human CD8 T cells associated with optimized ligand rigidity for TCR binding and not with changes in TCR:pMHC off-rate kinetics.	Sep 2020
I03-Macromolecular Crystallography I04-Macromolecular Crystallography	CD4+ T cells recognize conserved influenza a epitopes through shared patterns of V-gene usage and complementary biochemical features Alexander Greenshields-Watson , Meriem Attaf , Bruce J. Maclachlan , Thomas Whalley , Cristina Rius , Aaron Wall , Angharad Lloyd , Hywel Hughes , Kathryn E. Strange , Georgina H. Mason , Andrea J. Schauenburg , Sarah L. Hulin-Curtis , James Geary , Yuan Chen , Sarah N. Lauder , Kathryn Smart , Dhanasekaran Vijaykrishna , Miguel L. Grau , Mikhail Shugay , Robert Andrews , Garry Dolton , Pierre J. Rizkallah , Awen M. Gallimore , Andrew K. Sewell , Andrew J. Godkin , David K. Cole Cell Reports 32 DOI: 10.1016/j.celrep.2020.107885 Diamond Proposal Number(s): [10462, 14843] Open Access Show Abstract ▼ Abstract: T cell recognition of peptides presented by human leukocyte antigens (HLAs) is mediated by the highly variable T cell receptor (TCR). Despite this built-in TCR variability, individuals can mount immune responses against viral epitopes by using identical or highly related TCRs expressed on CD8+ T cells. Characterization of these TCRs has extended our understanding of the molecular mechanisms that govern the recognition of peptide-HLA. However, few examples exist for CD4+ T cells. Here, we investigate CD4+ T cell responses to the internal proteins of the influenza A virus that correlate with protective immunity. We identify five internal epitopes that are commonly recognized by CD4+ T cells in five HLA-DR1+ subjects and show conservation across viral strains and zoonotic reservoirs. TCR repertoire analysis demonstrates several shared gene usage biases underpinned by complementary biochemical features evident in a structural comparison. These epitopes are attractive targets for vaccination and other T cell therapies.	Jul 2020

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