Journal
BLOOD ADVANCES
Volume 2, Issue 23, Pages 3506-3514Publisher
AMER SOC HEMATOLOGY
DOI: 10.1182/bloodadvances.2017014639
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Funding
- Princess Margaret Cancer Foundation
- Canada Research Chairs program
- Canada Foundation for Innovation, Leaders Opportunity Fund, CFI [32383]
- Ontario Ministry of Research and Innovation, Ontario Research Fund Small Infrastructure Program
- CIHR Fellowship
- Terry Fox Foundation-CIHR Strategic Health Research Training Fellowship
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Mature T-cell lymphomas consisting of an expanded clonal population of T cells that possess common rearrangements of the T-cell receptor (TCR) encoding genes can be identified and monitored using molecular methods of T-cell repertoire analysis. We have developed a hybrid-capture method that enriches DNA sequencing libraries for fragments encoding rearranged TCR genes from all 4 loci in a single reaction. We use this method to describe the TCR repertoires of 63 putative lymphoma clinical isolates, 7 peripheral blood mononuclear cell (PBMC) populations, and a collection of tumor infiltrating lymphocytes. Dominant Variable (V) and Joining (J) gene pair rearrangements in cancer cells were confirmed by polymerase chain reaction (PCR) amplification and Sanger sequencing; clonality assessment of clinical isolates using BIOMED-2 methods showed agreement for 73% and 77% of samples at the beta and gamma loci, respectively, whereas beta locus V and J allele prevalence in PBMCs were well correlated with results from commercial PCR-based DNA sequencing assays (r(2) = 0.94 with Adaptive ImmunoSEQ, 0.77-0.83 with Invivoscribe LymphoTrack TRB Assay). CapTCR-seq allows for rapid, high-throughput and flexible characterization of dominant clones within TCR repertoire that will facilitate quantitative analysis of patient samples and enhance sensitivity of tumor surveillance over time.
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