GPA33 Marker Isolation of Stable Regulatory T Cells
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Solution Overview
Problem
Current methods for isolating and purifying stable regulatory T cells (Tregs) are inadequate, leading to contamination with conventional T cells and unstable iTregs, which can cause harmful immune responses in therapeutic applications.
Innovation Solution
The use of GPA33 as a marker to selectively enrich and isolate CD4+ T cells with high expression of GPA33, allowing for the purification of stable regulatory T cells (tTregs) while minimizing conventional T cells and unstable iTregs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If current isolation methods based on CD25 and CD127 markers are used, then Treg cells can be isolated, but the purity is insufficient due to contamination with conventional T cells and unstable iTregs
Solution Approach 1:
The patent changes the selection parameters from conventional markers (CD25+, CD127-) to a new marker profile (GPA33+, CD161+). This parameter change enables discrimination between stable tTregs and unstable iTregs, achieving >90% purity while maintaining population stability during expansion.
Solution Approach 2:
The patent extracts the unstable iTreg population from the mixed Treg preparation by identifying them as GPA33-CD161- cells. This extraction leaves behind a purified population of stable tTregs that are GPA33+CD161+, eliminating the harmful contaminating cells.
2Productivity
If Tregs are expanded in vitro without selective pressure, then expansion is easier, but contaminating Tconvs overgrow Tregs
Solution Approach 1:
The patent performs preliminary purification using GPA33 and CD161 markers before expansion to remove all contaminating Tconvs and unstable iTregs. This preliminary action ensures that only stable tTregs are present for expansion, eliminating the need for rapamycin and preventing overgrowth issues.
Solution Approach 2:
The patent uses GPA33 and CD161 markers as intermediaries to selectively identify and isolate stable tTregs. These markers serve as mediators that enable precise separation of stable Tregs from unstable cells, allowing clean expansion without additional drugs.
3Manufacturing precision
If rapamycin is used during expansion, then Treg purity is maintained, but expansion rate is suppressed and functional properties are altered
Solution Approach 1:
The patent converts the harmful effect of requiring rapamycin (which suppresses expansion and alters function) into a benefit by using GPA33/CD161 markers to achieve purity through selective isolation. This eliminates the need for rapamycin while maintaining both purity and natural expansion rates.
Solution Approach 2:
The stable tTregs naturally express high levels of GPA33 and CD161, allowing them to self-select and self-purify during expansion. This self-service mechanism maintains population purity without requiring external drugs like rapamycin, preserving both expansion rate and functional properties.
Data Source
AI summary
The invention relates to isolated populations of CD4+GPA33high regulatory T cells, to methods for detecting and/or isolating CD4+GPA33high cells, methods for enriching for CD4+GPA33high cells and to uses of CD4+GPA33high cells and populations of such cells. The invention further relates to methods for classifying an individual suffering from cancer and typing a tumor sample as well as for determining the etiology of autoimmune diseases.