Lymphocyte Replicative History Detection via Signal Joint Analysis
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Solution Overview
Problem
Current methods struggle to accurately monitor the efficacy of bone marrow transplantation by distinguishing between newly produced and old B cells, as existing techniques cannot discern the origin of B cells post-transplantation, leading to difficulties in assessing the regeneration of the precursor B-cell compartment.
Innovation Solution
A method involving the detection of signal joint and coding joint nucleotide sequences on extrachromosomal circular excision products allows discrimination between immature and mature B cells by quantifying the ratio of these sequences, which increases with each cell division, thereby determining the replicative history of lymphocytes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to monitor B cells post-transplantation, then the overall B-cell population can be measured, but the origin of B cells (newly produced vs. old) cannot be distinguished
Solution Approach 1:
The patent extracts and detects extrachromosomal circular excision products (BRECs) that contain signal joint nucleotide sequences as markers for recently produced B cells. By isolating and quantifying these specific molecular structures, the method separates the information about newly produced cells from the total B-cell population, enabling origin discrimination without requiring complex genomic analysis of all B cells.
Solution Approach 2:
The patent uses extrachromosomal circular excision products as intermediary markers to indirectly determine the replicative history of B cells. Instead of directly analyzing the complex genomic rearrangement history of each B cell, the method detects these excision products that serve as molecular fossils recording the gene rearrangement events, thereby mediating the information transfer from cellular replicative history to measurable molecular signals.
2Measurement precision
If gene rearrangement analysis is performed on all B cells, then comprehensive information is obtained, but the complexity and cost of analysis increases
Solution Approach 1:
The patent extracts and detects only the relevant signal joint nucleotide sequences present on extrachromosomal circular excision products, rather than performing comprehensive gene rearrangement analysis on all B cell genomes. This selective extraction approach focuses the analysis on the specific molecular markers that indicate recent production, significantly reducing the complexity and cost while maintaining precise replicative history determination.
Solution Approach 2:
The patent utilizes extrachromosomal circular excision products as disposable molecular markers that are easily detectable and do not require complex, expensive analysis infrastructure. These BRECs serve as simple, readily detectable proxies for complex cellular history, enabling precise measurement without the need for sophisticated sequencing or genomic analysis capabilities.
3Measurement precision
If the ratio of signal joint to coding joint sequences is quantified, then the replicative history can be determined, but the detection and quantification process becomes more complex
Solution Approach 1:
The patent employs extrachromosomal circular excision products as intermediary carriers that simultaneously contain both signal joint and coding joint nucleotide sequences. By detecting the ratio of these sequences within the same molecular structure (BREC), the method simplifies the measurement process compared to analyzing separate genomic loci, as the excision product itself serves as the template for detecting both sequence types in a single analytical framework.
Data Source
AI summary
The invention relates to the field of immunology and immunodiagnostics. Provided is a method for determining the replicative history of a lymphocyte, preferably a B cell, the method comprising detecting a signal joint nucleotide sequence on an extrachromosomal circular excision product in the lymphocyte, wherein the excision product is deleted from a chromosome to give a chromosomal-coding joint nucleotide sequence, wherein the coding joint is retained in the chromosome, and detecting the coding joint nucleotide sequence in the lymphocyte. Also provided are primers, probes and a control cell for use in a method of the invention. A method provided herein is among others advantageously used to assess recovery of the precursor B-cell compartment, for example, in a patient following bone marrow transplantation