Immune Cell Detection via CCR6 BLR1 Methylation
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Solution Overview
Problem
Current methods for identifying and distinguishing between stably and transiently activated immune cells, particularly T cells, are inadequate, as they fail to differentiate between cells that permanently express chemokine receptors like CCR6 and those that only express them temporarily, which is crucial for diagnosing and treating conditions such as rheumatoid arthritis and autoimmune diseases.
Innovation Solution
Analyzing the methylation status of specific CpG positions in the CCR6 and BLR1 genes using DNA methylation analysis to identify stably activated immune cells, as demethylation indicates stable activation and long-term expression of these receptors, allowing for the differentiation between transient and stable immune cell populations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current methods are used to identify immune cells, then the identification process is simple, but the ability to differentiate between stably and transiently activated cells is insufficient
Solution Approach 1:
The patent changes the detection parameter from protein expression levels (flow cytometry) or RNA expression (qPCR) to DNA methylation status. Methylation status is a stable epigenetic mark that persists through cell division, enabling differentiation between stable and transient activation. This parameter change resolves the contradiction by providing superior differentiation precision while maintaining reasonable method complexity through established methylation analysis techniques.
Solution Approach 2:
The patent introduces DNA methylation analysis as an intermediary marker to indirectly assess stable immune cell activation. Instead of directly measuring protein expression dynamics, the method uses methylation status of activation-associated genes as a stable surrogate marker. This intermediary approach enables precise differentiation without requiring complex longitudinal tracking of protein expression.
2Reliability
If protein expression analysis is used, then the method is straightforward, but it cannot distinguish stable from transient activation
Solution Approach 1:
The patent transitions from measuring transient protein expression levels to measuring stable DNA methylation patterns. Methylation status provides a permanent record of activation history that is retained through cell division, preserving temporal information about stable versus transient activation. This parameter change resolves the contradiction by determining activation status with higher reliability while preventing loss of temporal information.
Solution Approach 2:
The patent applies preliminary epigenetic marking during activation that persists long-term. DNA methylation changes occur during activation and are maintained as a molecular memory, allowing retrospective determination of activation stability. This preliminary action embedded in the epigenome enables reliable distinction between stable and transient activation without requiring continuous monitoring.
3Measurement precision
If methylation analysis is performed on multiple CpG positions, then the identification accuracy improves, but the analysis time increases
Solution Approach 1:
The patent segments the CCR6 and BLR1 genes into multiple CpG islands and analyzes methylation status at specific representative positions within each island. This segmentation approach maintains high identification accuracy by capturing the overall methylation pattern through strategic sampling of key positions, while reducing total analysis time compared to sequencing the entire gene regions.
Solution Approach 2:
The patent performs partial methylation analysis by focusing on selected CpG positions within CpG islands rather than analyzing every possible position in the gene. This partial action approach provides sufficient precision for cell identification by targeting informative regions, while significantly reducing analysis time and complexity compared to comprehensive gene-wide analysis.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method provides a reliable tool for identifying stable populations of immune cells, enabling improved diagnosis and treatment of autoimmune diseases by distinguishing between stably and transiently activated cells, thereby targeting the true immune response effectively.
Implementation Method 1
analysing the methylation status of at least one CpG position in the gene CCR6 and/or BLR1
Data Source
AI summary
The present invention relates to a method, in particular an in vitro method, for identifying certain immune cells of a mammal, comprising analysing the methylation status of at least one CpG position in the gene CCR6 and/or BLR1 or an orthologous or paralogous gene thereof, and the use of DNA-methylation analysis of the genes of the proteins CCR6 and/or BLR1 for a detection and quality assurance and control of certain immune cells. In particular, the present invention relates to analysing the methylation status of at least one CpG position in the gene CCR6 in T cells. Furthermore, the present invention relates to a kit for performing the above methods, as well as to respective uses.