Breg Cell Expansion via CD40 Agonist and Cytokines
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Solution Overview
Problem
The rarity and difficulty in characterizing human B regulatory cells (Breg) in peripheral blood hinder their clinical application, particularly in immune-mediated disorders and transplantation, due to their limited availability and complex characterization.
Innovation Solution
Ex vivo expansion of human B cells with a phenotype of CD19+CD73−CD71+CD25+TIM-1+, using a method involving isolation, culturing with a CD40 agonist and cytokines, and harvesting to produce a population with enhanced regulatory function, which can be used in pharmaceutical compositions for immune modulation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If human Breg cells are used for clinical therapy, then immune-mediated disorders can be treated, but the rarity and difficulty in characterizing these cells in peripheral blood hinder their availability and application
Solution Approach 1:
The patent performs preliminary characterization and expansion of human Breg cells in vitro before clinical application. By isolating and expanding Breg cells from peripheral blood mononuclear cells in a controlled laboratory setting, the patent overcomes the rarity issue and establishes a reproducible source for clinical therapy, directly addressing the availability barrier.
Solution Approach 2:
The patent defines and utilizes specific phenotypic parameters (CD19+CD73−CD71+CD25+TIM-1+) to identify and characterize human Breg cells. By establishing this marker profile, the patent transforms the characterization difficulty into a measurable criterion, enabling reliable identification and quantification of Breg cells for clinical purposes.
2Reliability
If human Breg cells are expanded ex vivo, then their regulatory function can be enhanced for therapy, but the complexity of isolation and culturing procedures increases
Solution Approach 1:
The patent optimizes culturing parameters including cytokine combinations (IL-2, IL-4, IL-10), culture duration (3-7 days), and cell density to maximize Breg cell expansion and regulatory function. By systematically adjusting these parameters, the patent achieves reliable enhancement of Breg functionality while maintaining a manageable protocol complexity.
Solution Approach 2:
The patent employs autologous Breg cell expansion where the cells are cultured with their own stimulation factors and cytokines, allowing them to self-regulate and maintain their regulatory function. This self-service approach simplifies the protocol by eliminating the need for complex external stimulation systems while ensuring functional reliability.
3Measurement precision
If TIM-1 expression is used to characterize Breg cells, then regulatory function can be identified, but the specificity and sensitivity of this marker in peripheral blood remains limited
Solution Approach 1:
The patent combines multiple markers (CD19, CD73, CD71, CD25, TIM-1) into a composite phenotypic profile to characterize human Breg cells. This merging of markers creates a more specific and sensitive identification system, where the combination of markers provides synergistic detection accuracy that individual markers cannot achieve alone, directly resolving the sensitivity limitation in peripheral blood.
Data Source
AI summary
The present invention relates to an expanded population of human Breg cells having the phenotype CD19+CD73−CD71+CD25+TIM-1+ and methods for producing the cell population of the invention. The invention also relates to pharmaceutical compositions comprising the cell populations of the invention and their use in the treatment of immune-mediated disorders.