Molecular T-cell Receptor Analysis

Location: Thomas Building, DE-550
Contact phone: (206) 667-6455
Contact fax: (206) 667-6845
Contact e-mail: jcao@fredhutch.org

TCR Spectratyping

Spectratyping is a set of multiplex RT-PCR reactions that amplifies all T-cell receptor beta chain families to reveal the clonal length polymorphism in the CDR3 region. This technique is used to access clonal composition in T-cell populations. Specifically, it is often used to determine clonal expansion, contraction and deletion during immune response and reconstitution. TCR spectratyping can be performed on PBMC samples or FACS-sorted cell subsets.

Clonal TCR and CAR-T analysis and tracking by real-time PCR

The unique feature of each T-cell clone is found in the recombined T cell receptor sequences. Immune Monitoring can identify the clonal TCR beta chain sequence by RT-PCR and direct sequencing of the clonal cell sample. The clonal TCRB sequence can be cloned and used for qPCR analysis.

With knowledge of the clonal TCR sequence, specific primers and a probe for quantitative real-time PCR analysis can be designed, allowing the quantitation of the clonal T cells in a cell population. We provide this service in the setting of T cell adoptive transfers in immune therapy trials of cancer patients. Custom designed qPCR assays are also provided for CAR-T cell therapy trials.

TREC assay

TREC is a quantitative real-time PCR analysis of the TCR excision circle, a circular DNA fragment excised from the genome as a result of the TCR rearrangement in the thymus. This circular DNA is not capable of replication and is therefore diluted as T cells divide in the periphery. The relative quantity of the TREC copy numbers provides a measurement of thymic output of naive T cells.

For any of the above assays please use iLab to request the service.