Tr1 Cell Isolation via Composite Marker Segmentation
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Solution Overview
Problem
The lack of specific biomarkers for T regulatory type 1 (Tr1) cells limits their study and clinical application, particularly in identifying and purifying Tr1 cells for therapeutic use, as existing markers are not selective and hinder the development of Tr1 cell-based clinical approaches.
Innovation Solution
The identification of specific cell surface markers such as CD4, CD49b, and LAG-3, with optional CD226 expression, allows for the selection, enrichment, isolation, and purification of Tr1 cells, enabling their administration for therapeutic purposes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If existing cell surface markers are used for Tr1 cell identification, then cell selection can be performed, but the markers are not selective enough and cannot effectively purify Tr1 cells
Solution Approach 1:
The patent segments the Tr1 cell identification process into multiple distinct marker evaluations (CD4, CD49b, LAG-3, CD226) rather than relying on a single marker. This segmentation allows for progressive filtering and purification of Tr1 cells through sequential gating strategies in flow cytometry, achieving both precision and practical feasibility
Solution Approach 2:
The patent employs a composite marker profile consisting of multiple cell surface markers (CD4+ CD49b+ LAG-3+ CD226+) to identify Tr1 cells. This composite approach combines the advantages of each individual marker to create a highly specific identification signature that overcomes the limitations of single-marker systems
2Manufacturing precision
If multiple cell surface markers are used for Tr1 cell purification, then purification quality improves, but the process complexity increases
Solution Approach 1:
The patent performs preliminary enrichment of Tr1 cells using easily accessible markers like CD4 before applying more complex multi-marker sorting strategies. This preliminary action simplifies the overall process by reducing the complexity of the sample before subjecting it to more rigorous purification protocols
Solution Approach 2:
The patent employs dynamic gating strategies in flow cytometry where the analysis and selection criteria are adjusted at different stages of the purification process. The methodology allows for flexible modification of marker combinations and gating thresholds based on sample characteristics and desired purification levels
3Adaptability or versatility
If specific Tr1 cell markers are identified, then clinical application and therapy development are facilitated, but the ability to transfer pure Tr1 cell populations remains limited
Solution Approach 1:
The patent incorporates feedback mechanisms through sequential marker evaluation and gating strategies where the identification of cells at one marker level informs subsequent sorting decisions. This feedback loop ensures that only cells meeting all criteria (CD4+ CD49b+ LAG-3+ CD226+) are selected, maintaining population purity while enabling clinical-scale preparations
Data Source
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AI summary
The invention is based upon the discovery that T regulatory type 1 (Trl) cells express particular cell surface markers that allow for their selection, enrichment, isolation, purification and administration. The ability to use the particular markers described herein to select, enrich, isolate, purity and administer Tr1 cells allows for improved methods of Tr1 therapies for treating a wide variety of diseases and disorders.