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Determination of thymic function directly from peripheral blood: a validated modification to an established method

Lookup NU author(s): Emeritus Professor Mike Sir Michael Rawlins, Dr Angela Patterson, Dr Arthur PrattORCiD, Dr Matthew Jefferson, Professor John IsaacsORCiD

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Abstract

The thymus contributes naive, self MHC reactive, self tolerant T cells to the peripheral immune system throughout life, albeit with a log-linear decline with age. Quantification of thymic function is clinically relevant in the setting of lymphoablation, but a phenotypic marker distinguishing recent thymic emigrants from long lived naive T cells remains elusive. T cell receptor excision circles (TREC) are present in thymocytes exiting the thymus and quantification of the most frequent of these, the deltarec-psiJalpha rearrangement has been widely used as a measure of recent thymic function. However, interpretation of results presented as TREC per cell has been criticised on the basis that extra-thymic cellular proliferation impacts on peripherally determined TREC numbers. TREC/ml is now considered to be more representative of thymic function than TREC/cell, especially where significant cellular proliferation occurs (e.g. during reconstitution following stem cell transplantation). Here we describe the validation of a novel variation to the established assay, directly quantifying TREC/ml from 300 microl whole blood. We show the assay to be reproducible, robust and stable longitudinally and we show equivalence of performance when compared with more standard assays. This assay particularly lends itself to the measurement of thymic function in children and where monitoring clinical variables is limited by tissue availability.

Publication metadata

Author(s): Lorenzi AR, Patterson AM, Pratt A, Jefferson M, Chapman CE, Ponchel F, Isaacs JD

Publication type: Article

Publication status: Published

Journal: Journal of Immunological Methods

Year: 2008

Volume: 339

Issue: 2

Pages: 185-94

Print publication date: 01/12/1931

ISSN (print): 0022-1759

ISSN (electronic): 1872-7905

Publisher: Elsevier

URL: http://dx.doi.org/10.1016/j.jim.2008.09.013

DOI: 10.1016/j.jim.2008.09.013

Notes: Arthritis Research Campaign/United Kingdom Journal Article Research Support, Non-U.S. Gov't Netherlands

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