T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones

Theaker, Sarah M.; Rius, Cristina; Greenshields‐Watson, Alexander; Lloyd, Alun G.; Trimby, Andrew; Fuller, Anna; Miles, John J.; Cole, David K.; Peakman, Mark; Sewell, Andrew K.; Dolton, Garry

doi:10.1016/j.jim.2016.01.014

Cited by 24 publications

(23 citation statements)

References 37 publications

(40 reference statements)

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“…T-cell clones, DCD10, and ILA1 were created ( 30 , 31 ) and passaged as previously described ( 32 ). Briefly, clones were expanded with irradiated (3,100 Gy) PBMCs from three donors in R10 [RPMI 1640 supplemented with 10% FBS, 100 U/ml penicillin, 100 µg/ml streptomycin, 1× MEM non-essential amino acid, 1 mM sodium pyruvate, 10 mM HEPES buffer (Life Technology)] with 20 IU/ml of IL-2 (Aldesluekin, Proleukin, Prometheus, San Diego, CA, USA) and 1 µg/ml of phytohaemaglutinin (Alere, Cheshire, UK).…”

Section: Methodsmentioning

confidence: 99%

Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation

et al. 2017

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Linking immunometabolic adaptation to T-cell function provides insight for the development of new therapeutic approaches in multiple disease settings. T-cell activation and downstream effector functions of CD4+ and CD8+ T-cells are controlled by the strength of interaction between the T-cell receptor (TCR) and peptides presented by human leukocyte antigens (pHLA). The role of TCR–pHLA interactions in modulating T-cell metabolism is unknown. Here, for the first time, we explore the relative contributions of the main metabolic pathways to functional responses in human CD4+ and CD8+ T-cells. Increased expression of hexokinase II accompanied by higher basal glycolysis is demonstrated in CD4+ T-cells; cytokine production in CD8+ T-cells is more reliant on oxidative phosphorylation. Using antigen-specific CD4+ and CD8+ T-cell clones and altered peptide ligands, we demonstrate that binding affinity tunes the underlying metabolic shift. Overall, this study provides important new insight into how metabolic pathways are controlled during antigen-specific activation of human T-cells.

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Section: Methodsmentioning

confidence: 99%

Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation

et al. 2017

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“…Additionally, several methods were developed to avoid a decline in their frequency upon expansion, due to overgrowth of non-reactive T cells. The first one, T cell library approach, was based on generation of more than a thousand parallel, small-scale T cell cultures, instead of expanding a bulk T cell culture from patient's peripheral blood [132,134]. In the second approach, peripheral blood T cells expressing PD-1 and 4-1BB were sorted, followed by limiting dilution and microwell culture [135] or bulk expansion [133].…”

Section: Selection Of Neoantigen-specific T Cells From Peripheral Bloodmentioning

confidence: 99%

Adoptive Cell Therapy—Harnessing Antigen-Specific T Cells to Target Solid Tumours

Chruściel

Urban-Wójciuk

Arcimowicz

et al. 2020

Cancers

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In recent years, much research has been focused on the field of adoptive cell therapies (ACT) that use native or genetically modified T cells as therapeutic tools. Immunotherapy with T cells expressing chimeric antigen receptors (CARs) demonstrated great success in the treatment of haematologic malignancies, whereas adoptive transfer of autologous tumour infiltrating lymphocytes (TILs) proved to be highly effective in metastatic melanoma. These encouraging results initiated many studies where ACT was tested as a treatment for various solid tumours. In this review, we provide an overview of the challenges of T cell-based immunotherapies of solid tumours. We describe alternative approaches for choosing the most efficient T cells for cancer treatment in terms of their tumour-specificity and phenotype. Finally, we present strategies for improvement of anti-tumour potential of T cells, including combination therapies.

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“…HLA A2 restricted clone InsB4 that recognises the HLVEALYLV peptide from the insulin chain (residues 10-18) (25) was grown from CD8 T-cells purified from a T1D patient using a T-cell library (26). Clone InsB6 was produced at the same time as InsB4 and subsequently found to express the same TCR.…”

Section: Cell Lines and T-cellsmentioning

confidence: 99%

“…T-cell clone GD.InsB4 (InsB4 from hereon) was generated from the blood of a patient with T1D using the T-cell libraries approach and the putative human insulin β chain 10−18 epitope, HLVEALYLV, as previously described (26), with peptide activation data shown in Figure 2A. This clone was initially shown to recognise K562 cells transduced with HLA A2 and preproinsulin (so-called surrogate human β-cells), which were not killed by a T-cell clone with a different specificity ( Figure 2B).…”

Section: T-cell Clone Insb4 Recognises a Bona Fide Hla A2-restricted mentioning

confidence: 99%

GPU-Accelerated Discovery of Pathogen-Derived Molecular Mimics of a T-Cell Insulin Epitope

et al. 2020

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The strong links between (Human Leukocyte Antigen) HLA, infection and autoimmunity combine to implicate T-cells as primary triggers of autoimmune disease (AD). T-cell crossreactivity between microbially-derived peptides and self-peptides has been shown to break tolerance and trigger AD in experimental animal models. Detailed examination of the potential for T-cell crossreactivity to trigger human AD will require means of predicting which peptides might be recognised by autoimmune T-cell receptors (TCRs). Recent developments in high throughput sequencing and bioinformatics mean that it is now possible to link individual TCRs to specific pathologies for the first time. Deconvolution of TCR function requires knowledge of TCR specificity. Positional Scanning Combinatorial Peptide Libraries (PS-CPLs) can be used to predict HLA-restriction and define antigenic peptides derived from self and pathogen proteins. In silico search of the known terrestrial proteome with a prediction algorithm that ranks potential antigens in order of recognition likelihood requires complex, large-scale computations over several days that are infeasible on a personal computer. We decreased the time required for peptide searching to under 30 min using multiple blocks on graphics processing units (GPUs). This time-efficient, cost-effective hardware accelerator was used to screen bacterial and fungal human pathogens for peptide sequences predicted to activate a T-cell clone, InsB4, that was isolated from a patient with type 1 diabetes and recognised the insulin B-derived epitope HLVEALYLV in the context of disease-risk allele HLA A * 0201. InsB4 was shown to kill HLA A * 0201 + human insulin producing β-cells demonstrating that T-cells with this specificity might contribute to disease. The GPU-accelerated algorithm and multispecies pathogen proteomic databases were validated to discover pathogen-derived peptide sequences that acted as super-agonists for the InsB4 T-cell clone. Peptide-MHC tetramer binding and surface plasmon resonance were used to confirm that the InsB4 TCR bound to the highest-ranked peptide agonists derived from infectious bacteria and fungi. Adoption of GPU-accelerated prediction of T-cell agonists has the capacity to revolutionise our understanding of AD by identifying potential targets for autoimmune T-cells. This approach has further potential for dissecting T-cell responses to infectious disease and cancer.

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T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones

Cited by 24 publications

References 37 publications

Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation

Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation

Adoptive Cell Therapy—Harnessing Antigen-Specific T Cells to Target Solid Tumours

GPU-Accelerated Discovery of Pathogen-Derived Molecular Mimics of a T-Cell Insulin Epitope

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