CD127-Based CD4+ Treg Isolation for FoxP3-Enriched Expansion
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Solution Overview
Problem
The lack of specific cell surface biomarkers for regulatory T cells (Tregs) in humans complicates their identification and isolation, limiting therapeutic interventions for autoimmune diseases and immune disorders, as existing markers like CD25 and CTLA-4 are also expressed on effector T cells, and intracellular markers like FoxP3 are not suitable for functional studies or expansion.
Innovation Solution
Utilizing CD127 as a biomarker, alone or in conjunction with other markers, to identify and isolate immunosuppressive FoxP3+ regulatory T cells, allowing for their enrichment and expansion, which can then be administered to modulate immune responses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If CD25 is used as a biomarker to identify Tregs, then Treg identification is simplified, but the marker is also expressed on effector T cells making specific identification problematic
Solution Approach 1:
The patent segments the Treg identification process by introducing CD127 as an additional marker that divides the T cell population into distinct subsets. By combining CD25 with CD127, the patent creates a multi-parameter gating strategy that segments effector T cells from regulatory T cells based on their differential expression patterns, resolving the ambiguity of using CD25 alone.
Solution Approach 2:
CD127 acts as an intermediary marker that mediates the distinction between Treg and effector T cells. While CD25 is expressed on both cell types, CD127 expression patterns differ, providing an intermediate layer of discrimination that enables specific Treg identification without relying solely on CD25.
2Measurement precision
If FoxP3 is used as a biomarker to identify Tregs, then Treg identification becomes more accurate, but the intracellular location makes it unsuitable for flow cytometry and functional studies
Solution Approach 1:
CD127 serves as an intermediary surface marker that bridges the gap between intracellular FoxP3 detection and functional studies. By using CD127 expression patterns (particularly CD127low/− cells) as a surrogate for FoxP3+ Tregs, the patent enables flow cytometry-based identification and isolation of regulatory T cells for functional analysis without requiring direct FoxP3 detection.
Solution Approach 2:
The patent substitutes the mechanical approach of intracellular FoxP3 staining with a surface-based CD127 flow cytometry approach. This replacement allows for easier cell isolation and functional studies while maintaining Treg identification accuracy through the correlation between CD127 expression and FoxP3 status.
3Measurement precision
If multiple markers like CD25, HLA-DR, CTLA-4, and GITR are used to define Tregs, then Treg identification becomes more specific, but the complexity of immunophenotyping increases
Solution Approach 1:
The patent extracts the essential Treg identification function from the complex panel of markers (CD25, HLA-DR, CTLA-4, GITR) and identifies CD127 as the critical discriminator. By focusing on CD127 expression patterns, particularly in conjunction with CD25, the patent simplifies the immunophenotyping process while maintaining the ability to accurately quantify Tregs.
Solution Approach 2:
CD127 serves multiple functions: it acts as a flow cytometry accessible surface marker, provides discrimination between Treg and effector cells, and enables cell isolation for functional studies. This multi-functionality replaces the need for multiple separate markers, simplifying the overall immunophenotyping approach.
4Measurement precision
If CD4+CD25hi Tregs are isolated using traditional markers, then Treg isolation is achieved, but the yield is limited and expansion is problematic
Solution Approach 1:
The patent segments the T cell population using CD127 expression to identify a broader Treg subset (CD4+CD25hiCD127low/−) that includes previously missed populations. This segmentation strategy reveals a larger Treg population that can be effectively isolated and expanded, overcoming the limitations of traditional CD25-based isolation.
Solution Approach 2:
The patent changes the parameter used for Treg identification from CD25 alone to a combination of CD25 and CD127 expression levels. This parameter change expands the identifiable Treg population to include CD127low/− cells, increasing the yield and enabling successful expansion while maintaining isolation purity.
Data Source
AI summary
The invention provides methods of isolating CD 127lo/− immunosuppressive regulatory T cells which can be greatly enriched for FoxP3, methods of expanding the isolated cells, pharmaceutical compositions of such cells, and methods of their use in the treatment of autoimmune and other immune system mediated disorders.