Determining T-cell specificity to understand and treat disease

Hadrup, Sine Reker; Newell, Evan W.

doi:10.1038/s41551-017-0143-4

Cited by 11 publications

(7 citation statements)

References 162 publications

(173 reference statements)

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“…5). These stable, ready-to-use MHC-I multimers, which can be converted to antigen-specific pMHC multimers in a single-step peptide addition, perfectly align with the state-of-the-art antigen-specific T cell detection platforms that require the rapid generation of large arrays of MHC multimers (6,20). We have shown the feasibility of using DS-A2 in both combinatorial color encoding strategies and DNA barcoding-based methods of T cell detection (Figs.…”

Section: Discussionmentioning

confidence: 78%

Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells

et al. 2019

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The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use in peptide-MHC multimer–based approaches to comprehensively analyze T cell immunity. To overcome this challenge, we demonstrate the use of functionally empty MHC class I molecules stabilized by a disulfide bond to link the α1 and α2 helices close to the F pocket. Peptide-loaded disulfide-stabilized HLA-A*02:01 shows complete structural overlap with wild-type HLA-A*02:01. Peptide-MHC multimers prepared using disulfide-stabilized HLA-A*02:01, HLA-A*24:02, and H-2Kb can be used to identify antigen-specific T cells, and they provide a better staining index for antigen-specific T cell detection compared with multimers prepared with wild-type MHC class I molecules. Disulfide-stabilized MHC class I molecules can be loaded with peptide in the multimerized form without affecting their capacity to stain T cells. We demonstrate the value of empty-loadable tetramers that are converted to antigen-specific tetramers by a single-step peptide addition through their use to identify T cells specific for mutation-derived neoantigens and other cancer-associated antigens in human melanoma.

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Section: Discussionmentioning

confidence: 78%

Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells

et al. 2019

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“…Furthermore, epitope spreading as evaluated here for CD8 + T cells may likewise occur for CD4 + T cells, and further insight into the relationship between CD4 + and CD8 + tumor-reactive T cells and the relevance for shared antigen recognition are critical aspects for addressing future improvements in immunotherapy. However, technical limitations still prohibit detailed epitope mapping of CD4 + NARTs, as conducted here for CD8 + NARTs ( 46 ).…”

Section: Discussionmentioning

confidence: 99%

Neoantigen-reactive CD8+ T cells affect clinical outcome of adoptive cell therapy with tumor-infiltrating lymphocytes in melanoma

Kristensen¹,

Heeke²,

Tvingsholm³

et al. 2022

Journal of Clinical Investigation

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BACKGROUND Neoantigen-driven recognition and T cell–mediated killing contribute to tumor clearance following adoptive cell therapy (ACT) with tumor-infiltrating lymphocytes (TILs). Yet how diversity, frequency, and persistence of expanded neoepitope-specific CD8 + T cells derived from TIL infusion products affect patient outcome is not fully determined. METHODS Using barcoded pMHC multimers, we provide a comprehensive mapping of CD8 + T cells recognizing neoepitopes in TIL infusion products and blood samples from 26 metastatic melanoma patients who received ACT. RESULTS We identified 106 neoepitopes within TIL infusion products corresponding to 1.8% of all predicted neoepitopes. We observed neoepitope-specific recognition to be virtually devoid in TIL infusion products given to patients with progressive disease outcome. Moreover, we found that the frequency of neoepitope-specific CD8 + T cells in TIL infusion products correlated with increased survival and that neoepitope-specific CD8 + T cells shared with the infusion product in posttreatment blood samples were unique to responders of TIL-ACT. Finally, we found that a transcriptional signature for lymphocyte activity within the tumor microenvironment was associated with a higher frequency of neoepitope-specific CD8 + T cells in the infusion product. CONCLUSIONS These data support previous case studies of neoepitope-specific CD8 + T cells in melanoma and indicate that successful TIL-ACT is associated with an expansion of neoepitope-specific CD8 + T cells. FUNDING NEYE Foundation; European Research Council; Lundbeck Foundation Fellowship; Carlsberg Foundation.

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“…Different agents such as fluorescent probes, DNA barcodes, and heavy metal tags are anchored to the multimer, aiding in the recognition of these antigen-specific T cells through various detection machinery like flow cytometry, sequencing, or mass cytometry respectively. [159,143] Through its enhanced TCR-MHC binding capacity, the MHC technique can reliably isolate and characterize the antigen-specific T cells. For example, the experiment can be performed by incubating T cells with fluorescent probed CD19 peptide-MHC tetramers to specifically tag the CD19+ CAR-expressing T cells and visualizing them via fluorescence-activated cell sorting flow analysis.…”

Section: T Cell Specificitymentioning

confidence: 99%

Materials for Improving Immune Cell Transfection

et al. 2021

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recognize and bind to tumor-associated antigens such as CD19 and epidermal growth factor receptor (EGFR) before releasing cytotoxic granules and effector cytokines to destroy cancer cells. [3] CAR-T therapy has shown remarkable success against CD19 expressing B cell hematological malignancies and, thus far, five products (Kymriah from Novartis; Yescarta and Tecartus from Kite Pharma/Gilead Sciences; and Breyanzi and Abecma from Bristol-Myers Squibb) have been approved by the US Food and Drug Administration (FDA) for clinical use. [4,5] Currently, there are at least 500 clinical trials registered on ClinicalTrials.gov database using engineered immune cells to treat various cancers including that of the brain, lungs, and skin.Despite the clinical success of CAR-T cells against B cell leukemia, CAR-T therapy still face tremendous challenges in manufacturing, safety, and affordability before it can become a viable clinical option. [6] One of the biggest technical difficulties is to transfect sensitive, primary T cells whereby biomolecules like oligonucleotides and proteins have to be delivered intracellularly and into the nuclei of cells. FDA-approved gold standard viruses and bulk electroporation suffer from low transfection efficiency while also perturbing the critical biological attributes of cells such as proliferation, metabolism, and gene expression. [7] This increases the time and costs for cell expansion, and without an efficient and cost-friendly transfection technology, the price (between USD 0.4-0.5 million per patient) for FDA-approved CAR treatments (Kymriah and Yescarta) will remain unaffordable and untimely. [8] The limitations in conventional transfection techniques have motivated the development of micro-and nanoplatforms such as microfluidics, nanoparticles, and high-aspect-ratio nanostructures to improve immune cell viability and throughput during transfection.Herein, we first provide an overview of CAR-T cell manufacturing, with emphasis on the science of transfection and limitations of traditional technology using viruses and bulk electroporation. There will also be discussion of other cell-based cancer immunotherapy using other types of promising immune cell types. Next, we describe emerging transfection platforms and companies that have been established to overcome gaps in CAR-T transfection. We then provide a list of assays constituting the polyfunctionalities of immune cells that we believe will help the field better assess the robustness and suitability of their transfection methods. Finally, we end off with existing challenges in CAR-T transfection and how overcoming these challenges can significantly enhance the clinical impact of CAR-T therapy.Chimeric antigen receptor T cell (CAR-T) therapy holds great promise for preventing and treating deadly diseases such as cancer. However, it remains challenging to transfect and engineer primary immune cells for clinical cell manufacturing. Conventional tools using viral vectors and bulk electroporation suffer from low efficiency while posing risks ...

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Determining T-cell specificity to understand and treat disease

Cited by 11 publications

References 162 publications

Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells

Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells

Neoantigen-reactive CD8+ T cells affect clinical outcome of adoptive cell therapy with tumor-infiltrating lymphocytes in melanoma

Materials for Improving Immune Cell Transfection

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