CD4+ T-Cell Subpopulation Measurement Using CXCR3 Gating
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Solution Overview
Problem
Existing methods for measuring specific cell subpopulations in CD4+ T cells, such as CCR4-CCR6+ cells, are inefficient and require specialized facilities and techniques, leading to variability and inconvenience in sample collection and processing.
Innovation Solution
The method involves separating CXCR3+/- cell groups to measure CCR4-CCR6+ cell subpopulations in CD4+ T cells, using preferred storage conditions for whole blood samples, and employing a kit with detecting agents for CD4, CCR4, and CCR6, allowing measurements to be taken at the collection site without immediate processing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to measure CCR4-CCR6+ cell subpopulations, then measurement can be performed, but the process requires specialized facilities (centrifuge, freezing equipment), multiple processing steps, and generates variability between personnel and institutions
Solution Approach 1:
The invention extracts and measures CXCR3 expression directly from whole blood samples without requiring separation of peripheral blood mononuclear cells (PBMCs). This eliminates the need for centrifuges, freezing equipment, and complex laboratory processing, while maintaining measurement accuracy through direct detection of CXCR3+ cells among CD4+ T cells in the original blood sample
Solution Approach 2:
The measurement method enables the blood collection venue to perform the measurement itself using only a flow cytometer, without needing to transport samples to specialized laboratories. The whole blood sample serves its own purpose as both collection medium and measurement substrate, eliminating dependency on external facilities and personnel expertise
2Reliability
If PBMCs are separated and frozen at the blood collection venue, then cell subpopulations can be measured, but the process requires transportation, storage facilities, and increases time and operational complexity
Solution Approach 1:
The invention performs the measurement on whole blood samples immediately or shortly after collection, before any separation or freezing is required. By using CXCR3 expression as the measurement target in intact whole blood, the method eliminates the need for preliminary PBMC separation and freezing steps, reducing processing time while maintaining measurement stability through proper sample handling protocols
3Difficulty of detecting and measuring
If multiple detecting agents are used to identify CCR4-CCR6+ cells, then specific cell subpopulations can be identified, but the process becomes more complex and requires more reagents and processing steps
Solution Approach 1:
The invention extracts the measurement to a single marker (CXCR3) that can be detected directly in whole blood, eliminating the need to combine multiple detecting agents for CCR4 and CCR6. This simplifies the detection process to using only anti-CD4 and anti-CXCR3 antibodies, reducing reagent complexity and processing steps while maintaining the ability to identify the desired cell subpopulation
Data Source
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AI summary
[Problem] To provide: a method for more efficiently measuring the relative amount of a specific cell subpopulation in a CD4+ T cell population; and a kit for the method. [Solution] The present invention provides a method for determining the relative amount of a CCR4-CCR6+ cell subpopulation in a CD4+ T-cell population in a sample derived from a subject, the method comprising: a step for selecting a CXCR3- cell subpopulation in the CD4+ T-cell population; a step for setting a boundary between a CCR4- cell subpopulation and a CCR4+ cell subpopulation in the CXCR3- cell subpopulation; and a step for measuring the relative amount of the CCR4- CCR6+ cell subpopulation in the CD4+ T-cell population by using the boundary.