Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 352, Issue 1-2, Pages 111-117Publisher
ELSEVIER
DOI: 10.1016/j.jim.2009.11.007
Keywords
Sj-TREC; D beta J beta-TREC; TREC ratio; Intrathymic proliferation; HIV; Human thymic function
Categories
Funding
- Fondo de Investigaciones Sanitarias [FIS06/00176, CP08/00172]
- Fundacion la para la Investigacion y la Prevencion del SIDA en Espana (FIPSE) [12481/05, 36624/06]
- Redes Tematicas de Investigacion en SIDA [ISCIII RETIC RD06/0006/0021]
- Redes Tematicas de Cardiovascular [ISCIII RECAVA RD06/0014]
- Proyecto de Excelencia, Consejeria de Innovacion, Ciencia y Empresa [P06-CTS-01579]
- Consejeria de Salud, Servicio Andaluz de Salud [156/2006, PI0366/07]
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Current techniques to peripherally assess thymic function are: the signal-joint T-cell receptor excision circle (sj-TREC) level measurement and the naive T cell and CD31 + TREC-rich subset determination. However, all of them are indirect approaches and none could be considered a direct recent thymic emigrant (RTE) marker. To overcome their limitations, Dion et al. (2004) described the sj/beta-TREC ratio that allows the peripheral quantification of the double negative to double positive intrathymic proliferation step. Nevertheless, the protocol described is expensive, sample and time-consuming, thus, limiting its usefulness. In this study, we describe a simplified protocol that reduces from 33 to 9 the amount of PCR reaction needed but maintaining the sensitivity and reproducibility of the original technique. In addition, we corroborated the effectiveness of our technique as an accurate thymic output-related marker by correlating the peripheral sj/beta-TREC ratio with a direct measurement of thymic function as the percentage of double positive thymocytes (r = 0.601, p < 0.001). (C) 2009 Elsevier B.V. All rights reserved.
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