Anucleate Cell Intracellular Antigen Delivery via Microfluidic Constriction
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Solution Overview
Problem
Current methods for delivering antigens into red blood cells to induce immune tolerance are limited by the irregular shape and transcriptional inactivity of red blood cells, making standard transfection techniques ineffective and requiring chemically modified antigens with limited surface area for loading, which can be immunogenic.
Innovation Solution
Passing a cell suspension containing an anucleate cell, such as a red blood cell, through a constriction to deform the cell and facilitate the entry of an antigen or tolerogenic factor into the cell, thereby processing the antigen in a tolerogenic environment to suppress the immune response.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If standard transfection techniques are used to deliver antigens into red blood cells, then the process is simple and straightforward, but the technique is ineffective due to the irregular shape and transcriptional inactivity of red blood cells
Solution Approach 1:
The patent changes the physical state and mechanical properties of red blood cells by applying shear stress through microfluidic constrictions, transforming them from a static, irregular biconcave shape to a dynamic, deformable state that enables intracellular antigen delivery. This mechanical parameter change allows standard antigens to be delivered without chemical modification.
Solution Approach 2:
The patent replaces standard biochemical transfection methods with a mechanical delivery system using microfluidic constrictions. The mechanical shear stress generated in the constriction zone physically forces antigens into the cell membrane and cytoplasm, substituting chemical transfection mechanisms with a purely mechanical approach that overcomes the transcriptional inactivity and irregular shape limitations.
2Quantity of substance
If chemically modified antigens are conjugated to the surface of red blood cells, then antigen delivery is achieved, but the surface area is limited and the antigens become immunogenic
Solution Approach 1:
The patent transitions from two-dimensional surface conjugation to three-dimensional intracellular delivery. By delivering antigens into the cell interior through microfluidic constrictions, the available loading capacity expands from the limited cell surface area to the entire cell volume, while the antigens remain in their native, non-immunogenic form.
Solution Approach 2:
The patent extracts antigens from the problematic surface conjugation environment and delivers them directly into the intracellular space. This extraction from the surface interface eliminates the need for chemical modifications that cause immunogenicity, while maximizing the available loading volume within the cell cytoplasm.
3Reliability
If red blood cells are manipulated to associate antigenic material on their surface, then antigen presentation is achieved, but the manipulation is challenging due to cell shape and lack of nucleus
Solution Approach 1:
The patent enables red blood cells to self-deform and self-uptake antigens as they naturally flow through microfluidic constrictions. The cell's inherent deformability, when combined with the controlled shear stress in the constriction zone, allows automatic intracellular delivery without requiring external manipulation techniques that would be complicated by the cell's irregular shape and anucleate structure.
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 effectively delivers antigens into red blood cells, inducing a powerful immunosuppressive response and promoting immune tolerance, which is antigen-specific or non-specific, depending on the application.
Implementation Method 1
passing a cell suspension comprising an anucleate cell through a constriction, wherein said constriction deforms the cell thereby causing a perturbation of the cell such that an antigen enters the anucleate cell
Data Source
AI summary
The present invention provides methods for inducing tolerance and/or suppressing an immune response to an antigen by passing a cell suspension containing an anucleate cell through a constriction, wherein the constriction deforms the cell thereby causing a perturbation of the cell such that an antigen and/or tolerogenic factor enters the cell. In some embodiments, the anucleate cell is delivered to an individual and the antigen is delivered to and processed in a tolerogenic environment to induce tolerance and/or suppress an immune response to the antigen.