Overview
This report outlines a phenotyping assay for analyzing fresh whole blood to identify major leukocyte populations. The procedure is designed for multicenter clinical trials, ensuring consistency and accuracy in leukocyte enumeration.
Key Study Components
Area of Science
- Immunology
- Clinical Trials
- Flow Cytometry
Background
- Leukocyte populations play a crucial role in immune response.
- Accurate enumeration is essential for monitoring interventions in clinical trials.
- Standardized procedures enhance data reliability across multiple sites.
- Fluorescent antibodies are utilized for specific cell population staining.
Purpose of Study
- To develop a standardized staining procedure for fresh whole blood.
- To enable accurate analysis of leukocyte populations in multicenter trials.
- To facilitate tracking of immune responses over time.
Methods Used
- Utilization of tru count tubes containing lyophilized pellets.
- Dissolution of pellets during sample preparation to release fluorescent beads.
- Staining with fluorescent antibodies targeting specific surface markers.
- Flow cytometry analysis to compare cell populations with fluorescent beads.
Main Results
- Successful enumeration of major innate and adaptive leukocyte populations.
- Establishment of a reliable method for tracking leukocyte concentrations.
- Demonstration of the procedure's applicability in multicenter clinical trials.
- Validation of flow cytometry as an effective analysis tool.
Conclusions
- The developed procedure is effective for leukocyte phenotyping.
- Standardization is critical for multicenter trial integrity.
- Future studies can build on this methodology for broader applications.
What is the purpose of the staining procedure?
The staining procedure is designed to accurately enumerate leukocyte populations in fresh whole blood for clinical trials.
How does the tru count tube work?
The tru count tube contains a lyophilized pellet that releases a known number of fluorescent beads during sample preparation.
What role do fluorescent antibodies play in this procedure?
Fluorescent antibodies stain specific surface markers on different cell populations, allowing for their identification and enumeration.
Why is flow cytometry used in this study?
Flow cytometry is used to analyze the stained samples and compare the number of cells to the fluorescent beads for accurate quantification.
What are the implications of this research?
This research provides a standardized method for leukocyte analysis, enhancing the reliability of data in multicenter clinical trials.
Who developed this procedure?
The procedure was developed by the HIV Vaccine Trials Network at Fred Hutchinson Cancer Research Center.
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