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Simultaneous Quantification of T-Cell Receptor Excision Circles (TRECs) and K-Deleting Recombination Excision Circles (KRECs) by Real-time PCR
Published on: December 6, 2014
A reliable and simplified sj/beta-TREC ratio quantification method for human thymic output measurement
Sara Ferrando-Martínez1, Jaime M Franco, Ezequiel Ruiz-Mateos
1Laboratory of Immunovirology, Biomedicine Institute of Seville, Service of Infectious Diseases, Virgen del Rocio University Hospital, Avda. Manuel Siurot s/n 41013 Seville, Spain.
Journal of Immunological Methods
|November 19, 2009
Summary
This study simplifies measuring thymic function by reducing PCR reactions for the sj/beta-TREC ratio. This cost-effective method accurately reflects recent thymic emigrant output.
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- Assessing thymic function peripherally often relies on indirect markers like signal-joint T-cell receptor excision circle (sj-TREC) levels.
- Existing methods lack direct measurement of recent thymic emigrants (RTEs) and can be complex or time-consuming.
- The sj/beta-TREC ratio offers a more direct measure of thymic output but the original protocol is resource-intensive.
Purpose of the Study:
- To develop a simplified and more efficient protocol for quantifying the sj/beta-TREC ratio.
- To establish a cost-effective and sensitive method for assessing thymic function.
- To validate the simplified sj/beta-TREC ratio as a reliable marker of thymic output.
Main Methods:
- Developed a streamlined protocol reducing PCR reactions from 33 to 9.
- Maintained the sensitivity and reproducibility of the original sj/beta-TREC ratio technique.
- Correlated the peripheral sj/beta-TREC ratio with the percentage of double positive thymocytes, a direct measure of thymic function.
Main Results:
- The simplified protocol significantly reduces the number of PCR reactions required for sj/beta-TREC ratio analysis.
- The method maintains high sensitivity and reproducibility compared to the original protocol.
- A strong correlation was observed between the peripheral sj/beta-TREC ratio and the percentage of double positive thymocytes (r=0.601, p<0.001), confirming its accuracy.
Conclusions:
- The simplified sj/beta-TREC ratio protocol offers a more practical and cost-effective approach to assessing thymic function.
- This optimized method provides a reliable peripheral marker for recent thymic emigrant output.
- The technique enhances the ability to monitor thymic activity and T-cell regeneration.
