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Updated: Aug 8, 2025

Examination of Thymic Positive and Negative Selection by Flow Cytometry
Published on: October 8, 2012
Evaluation of thymopoiesis in healthy Turkish children aged 0-6 years
Akif Kavgacı1, Deniz Bayrakoğlu2, Sevgi Köstel Bal2
1Department of Pediatrics, Ankara University Faculty of Medicine, Ankara.
Insights
This study establishes reference values for recent thymic emigrants (RTE) in Turkish children, crucial for diagnosing primary immunodeficiencies (PIDs) like SCID. These findings aid in early diagnosis and monitoring immune reconstitution.
Area of Science:
- Immunology
- Pediatrics
- Public Health
Background:
- Primary immunodeficiencies (PIDs) are a significant public health concern requiring early diagnosis and treatment.
- Severe combined immunodeficiency (SCID) involves defective T-cell development due to genetic mutations affecting thymopoiesis.
- Assessing thymopoiesis is critical for diagnosing SCID and other combined immune deficiencies (CIDs).
Purpose of the Study:
- To establish reference values for recent thymic emigrants (RTE) in healthy Turkish children (0-6 years).
- To evaluate thymopoiesis by measuring RTE (T lymphocytes expressing CD4, CD45RA, CD31).
- To provide a benchmark for diagnosing PIDs in regions lacking newborn screening.
Main Methods:
- Flow cytometry was used to measure RTE in the peripheral blood of 120 healthy Turkish children.
- Samples included cord blood and peripheral blood from infants and children aged 0-6 years.
- Analysis focused on absolute RTE counts and their relative ratios.
Main Results:
- RTE cell counts and ratios were highest in the first year of life, peaking at 6 months.
- RTE levels significantly decreased with age after birth.
- Absolute lymphocyte count (ALC) also showed a decreasing trend with age.
Conclusions:
- Normal thymopoiesis and RTE reference levels were established for healthy Turkish children (0-6 years).
- This data can aid in the early diagnosis and monitoring of immune reconstitution in PID patients, particularly SCID and CIDs.
- RTE measurement offers a fast, reliable marker for PIDs, especially where newborn screening (NBS) is unavailable.
Background:
Early diagnosis and effective treatment serve as life-saving procedures for primary immunodeficiencies (PIDs) which are very common and a major public health problem in Turkey. Severe combined immunodeficiency (SCID) is constitutively a T-cell defect in which naïve T-cell development is defective due to the mutations in genes responsible for the T cell differentiation and insufficient thymopoiesis. So, assessment of thymopoiesis is very important in the diagnosis of SCID and several combined immune deficiencies (CIDs).
Methods:
The purpose of this study is to examine thymopoiesis in healthy children via measurement of recent thymic emigrants (RTE); T lymphocytes that express CD4, CD45RA and CD31 to establish the RTE reference values in Turkish children. RTE were measured in the peripheral blood (PB) of 120 healthy infants and children between 0-6 years including cord blood samples, by flow cytometry.
Results:
The absolute count of RTE cells and their relative ratios were found to be higher during the first year of life, being highest at the 6th month and tending to decrease significantly by age following birth (p=0.001). In the cord blood group, both values were lower than those in the 6-month-old group. The absolute lymphocyte count (ALC) varying by age, was found to reduce to 1850/mm³ in 4-years and after.
Conclusions:
Here we evaluated normal thymopoiesis and established the normal reference levels of RTE cells in the peripheral blood of healthy children aged between 0-6 years. We believe that the collected data will contribute to early diagnosis and monitoring of immune reconstitution; serving as an additional fast and reliable marker for many PID patients especially for SCID including many other CIDs, especially in nations where newborn screening (NBS) via T cell receptor excision circles (TREC) has not yet become available.
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