Detection and quantification of bovine signal joint T-cell receptor excision circles

A signal joint (sj) T-cell receptor excision circle (TREC) is produced by T-cell receptor (TCR) gene rearrangements during alpha beta T-cell maturation in the thymus. sjTREC have been studied as a marker of thymic function in several spices. We designed specific primers for delta rec-psi J alpha sj region to identify the location of the bovine sjTREC region and determined the nucleotide sequence of the PCR product. The obtained sequences were subjected to a BLAST search, which identified a matching region. This matching region contained TCR delta genes and was identified on bovine chromosome 10. We also confirmed the polymorphism of the sj region by sequencing of 10 PCR products, and observed irregular insertion of bases in the delta rec-psi J alpha recombination signal sequence. We then developed a quantitative PCR (QPCR) assay for evaluation of sjTRECs level in order to evaluate bovine thymic function for application in the veterinary clinic. This QPCR assay specifically amplified the sj region of bovine sjTREC and could detected 10(1)-10(7) copy numbers of sjTRECs. Using this assay we found that the number of sjTRECs in peripheral blood mononuclear cells was less than 10% that of the thymus. (C) 2015 Elsevier B.V. All rights reserved.