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Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper (cTfh) Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry
Published on: January 7, 2019
Efficient characterization of the TCR repertoire in lymph nodes by flow cytometry
D Salameire1, Y Le Bris, B Fabre
1Université Joseph Fourier Grenoble 1, F-38000, France.
Insights
Flow cytometry analysis of T-cell receptor (TCR) Vbeta repertoires in lymph nodes reveals diverse T-cell populations. Findings suggest lymph node enlargement stems from increased circulating T-cell homing, not pathology.
Area of Science:
- Immunology
- Flow Cytometry
- T-cell Biology
Background:
- T-cell receptor (TCR) repertoire analysis via flow cytometry is crucial for understanding T-cell diversity and identifying reactive cells.
- Characterizing non-malignant T-lymphocytes in lymph nodes is essential for insights into their origin and function.
Purpose of the Study:
- To analyze the T-cell receptor (TCR) Vbeta repertoire of CD4+ and CD8+ T-lymphocytes in lymph nodes.
- To investigate the origin of non-malignant T-lymphocytes in lymph nodes.
- To establish reference values for TCR-Vbeta expression in lymph nodes.
Main Methods:
- Analysis of the TCR repertoire in 81 lymph node samples using four-color flow cytometry.
- Utilized a panel of 25 anti-Vbeta monoclonal antibodies to characterize T-cell populations.
- Compared TCR-Vbeta repertoire in lymph node T-cells with blood samples.
Main Results:
- Demonstrated a diversified TCR-Vbeta repertoire with low-level expansions in 53% of samples, involving most Vbeta families.
- Expansions were more frequent in CD8+ T-cells of older patients and not linked to specific pathologies.
- TCR-Vbeta repertoire in lymph nodes was highly similar to that in blood samples.
Conclusions:
- Lymph node enlargement in the studied cases is primarily attributed to increased homing of circulating T-cells.
- The findings provide valuable reference data for TCR-Vbeta expression in lymph nodes.
- This data can aid in the diagnosis of T-cell lymphoproliferative disorders.
Abstract:
Analysis of the T-cell receptor (TCR) repertoire by flow cytometry proved to be relevant for investigating T-cell diversity and detecting reactive cells in blood samples. We used this approach to characterize non-malignant T-lymphocytes in lymph nodes and give insights into their origin. The TCR repertoire of CD4+ and CD8+ T-cells from 81 lymph nodes was analyzed with a four-color flow cytometer using a wide panel of 25 anti-Vbeta monoclonal antibodies. Flow cytometry proved to be a useful and informative technique. We demonstrated a diversified TCR-Vbeta repertoire, and only low level expansions, in 53% of the samples. They involved nearly all Vbeta families, were more frequent in the CD8+ subset of older patients, but were not related to pathology. No evidence could be demonstrated in favor of stimulation by common antigens. Interestingly, the TCR-Vbeta repertoire proved to be very similar in lymph nodes and blood samples. Our results argue that in the cases studied, lymph node enlargement is mainly due to an increased homing of circulating T-cells. They also provide reference values for expression of 25 TCR-Vbeta in lymph nodes, which could serve as a basis for further applications in diagnosis of T-cell lymphoproliferative disorders.
