Performance Verification and Establishment of Reference Intervals for Dendritic Cells, γδT Cells, and Follicular
Insights
Standardized flow cytometry methods were developed for dendritic cells (DCs), gamma-delta T (γδT) cells, and T follicular helper (Tfh) cells. This study established performance verification and reference intervals for improved clinical application of these immune cell subsets.
Area of Science:
- Immunology
- Clinical Chemistry
- Cell Biology
Background:
- Dendritic cells (DCs), gamma-delta T (γδT) cells, and T follicular helper (Tfh) cells are crucial for immune responses.
- Flow cytometry (FCM) offers precise quantification but lacks standardization for clinical use.
- Standardized performance verification and reference intervals are needed for these immune cell subsets.
Purpose of the Study:
- To standardize peripheral DCs, γδT cells, and Tfh cells analysis using FCM.
- To establish performance verification and reference intervals for these immune cell subsets.
- To facilitate broader clinical application of FCM-based immune cell subset analysis.
Main Methods:
- Collected peripheral whole blood from healthy individuals.
- Measured frequencies of DCs, γδT cells, and Tfh cells and their subpopulations via FCM.
- Evaluated sample stability, reportable range, precision, and linearity based on best practices.
- Established reference intervals for the analyzed immune cell subsets.
Main Results:
- Determined sample stability: DCs (24h at 4°C), γδT cells (96h at 4°C/RT), Tfh cells (instability after 24h at 4°C/RT).
- Established reportable ranges: DCs (5000 cells), γδT cells (250 cells), Tfh cells (1000 cells).
- Confirmed intra-assay/interassay precision and linearity met acceptance criteria; established reference intervals.
Conclusions:
- Systematic performance verification and reference intervals were established for DCs, γδT cells, and Tfh cells via FCM.
- This standardization promotes reliable clinical application of these immune cell subset assays.
- The findings support improved diagnostic and monitoring capabilities for immune-related conditions.
Context.—:
Dendritic cells (DCs), gamma-delta T (γδT) cells, and T follicular helper (Tfh) cells are key players in immune responses and clinical surveillance. While flow cytometry (FCM) provides precise quantification of these subsets with their subpopulations, its clinical application remains limited, partly due to lack of standardized performance verification and reference intervals.
Objective.—:
To facilitate the standardization and clinical application of peripheral DCs, γδT cells, and Tfh cells with their subpopulations, based on FCM.
Design.—:
Peripheral whole blood from healthy individuals was collected to measure the frequency of DCs, γδT cells, and Tfh cells with their subpopulations via FCM. Sample stability, reportable range, intra-assay/interassay precision, and linearity were evaluated from best practice documents, followed by establishing reference intervals.
Results.—:
For sample stability, samples were stable up to 24 hours post collection at 4°C for DCs with their subpopulations; samples were stable up to 96 hours at both 4°C and room temperature for γδT cells with their subpopulations; samples showed instability after 24 hours at both 4°C and room temperature for Tfh cells with their subpopulations. For the reportable range, the minimum cell numbers of upper gate for target cells that could meet the detection requirements of DCs, γδT cells, and Tfh cells with their subpopulations were 5000, 250, and 1000. Both intra-assay/interassay precision and linearity fulfilled the acceptance criteria. Furthermore, we established the reference intervals for DCs, γδT cells, and Tfh cells with their subpopulations.
Conclusions.—:
We conducted systematic performance verification and established reference intervals for DCs, γδT cells, and Tfh cells with their subpopulations via FCM, promoting the standardization and clinical application of these assays.
