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Development of a TCR beta repertoire assay for profiling liquid biopsies from NSCLC donors

Cancer Drug Resistance 2020年第3期3卷 563-571页

作者： Leisa P.Jackson benjamin a.tjoa Hestia Mellert Gary A.Pestano Development Biodesix Inc.BoulderCO 80301USA. Cellero LLCBothellWashingtonWA 98021USA.

Aim:The aim of this study was to demonstrate the utility of T-Cell receptor beta(TCRβ)sequencing as a robust method for assessing T-cell repertoire changes in donors with non-small cell lung cancer(NSCLC).We further demonstrated the use of the assay by monitoring repertoire modulation in a defined model antigen system,cytomegalovirus(CMV).Methods:Peripheral blood mononuclear cells from four healthy donors were challenged with a 1-week exposure to whole-cell lysate from CMV-infected cells or CMVpp65495-503 peptide(NLVPMVATV).T-cell repertoire perturbations were assessed using the Oncomine TCR Beta-SR Assay and Ion GeneStudio S5 Plus Sequencer.A pp65 tetramer flow cytometry assay was used as an orthogonal method to assess clonal expansion of a subset of CMV-specific *** evaluation of the assay in peripheral blood lymphocytes from NSCLC donors,five whole blood specimens were evaluated using the same sequencing ***:The TCR beta assay identified 6,683-61,936 unique clones from 1-2 million reads per sample,and an average of 80%of the total reads were usable for TCR *** the NSCLC donors,TCR convergence and clonality values were consistent with published results and ranged 0.016-0.033 for convergence and 0.09-0.48 for *** the CMV study,TCR sequencing detected the expansion of a common family of clones in all 4 samples in response to antigen *** expansion corresponded to an increase in pp65 tetramer staining by flow *** TCR convergence scores ranged 0.009-0.041 and increased 5-fold in one sample as a result of pp65 antigen ***:The results of this study demonstrated the utility of profiling of the TCRβrepertoire in a model system and in donors with ***,we demonstrated the correlation between RNA-seq methods and protein-tetramer analysis using flow *** techniques represent an emerging solution that could complement other liquid and tissue diagnostic assays in the cl

关键词： TCRβ liquid biopsy Ion Torrent immunotherapy

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