One-Step CTL and Gamma Delta T Cell Culture Method
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Solution Overview
Problem
Current methods for culturing disease antigen-specific CTLs and γδT cells are inefficient and cumbersome, requiring multiple steps and varying culture conditions, making it difficult to obtain sufficient numbers of both cell types simultaneously for effective medical treatment.
Innovation Solution
A one-step culture method involving the addition of a disease antigen and an aminobisphosphonate, such as pamidronic acid, to peripheral blood mononuclear cells, which activates and proliferates γδT cells to function as antigen-presenting cells, inducing disease antigen-specific CTLs, thereby eliminating the need for separate cell separation and dendritic cell induction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If dendritic cells are used to induce disease antigen specific CTLs through multiple steps including monocyte separation and culture, then the ability to induce CTLs is improved, but the process complexity and time required increase significantly
Solution Approach 1:
The patent combines the functions of monocyte separation, dendritic cell induction, and CTL induction into a single integrated process using γδT cells as antigen-presenting cells. This eliminates the need for separate monocyte isolation and dendritic cell culture steps, directly resolving the contradiction between reliable CTL induction and process complexity.
Solution Approach 2:
The patent segments the traditional multi-step process by using γδT cells that can directly present antigens without requiring conversion from monocytes. This functional segmentation allows antigen-presenting cell activity to be achieved through a different cell type that naturally possesses this capability, simplifying the overall process.
2Reliability
If multiple cell types (CTLs and γδT cells) are cultured separately under different conditions, then each cell type can be optimized, but the overall productivity and efficiency decrease
Solution Approach 1:
The patent employs γδT cells that serve multiple functions: they act as antigen-presenting cells to induce CTLs while simultaneously being the target of antigen-specific activation. This multi-functionality allows a single cell type to fulfill roles that traditionally required multiple separate culture processes, thereby improving productivity without sacrificing functional optimization.
Solution Approach 2:
The patent merges the culture of γδT cells and CTL induction into a single co-culture system. By combining these processes, the system achieves both cell type optimization and improved productivity, as the γδT cells naturally present antigens to induce CTLs while being activated by the same antigen, creating a synergistic effect.
3Reliability
If γδT cells are activated with non-peptide antigens like alkylamines or bisphosphonates, then their antitumor activity is enhanced, but the purity and number of cells sufficient for treatment cannot be secured from small blood volumes
Solution Approach 1:
The patent combines γδT cell activation with CTL induction in a single process. The γδT cells are activated by non-peptide antigens to enhance their antitumor activity, while simultaneously serving as antigen-presenting cells to induce CTLs. This merging allows the system to achieve both high cell functionality and sufficient cell numbers from limited blood volumes.
Solution Approach 2:
The patent performs preliminary activation of γδT cells with non-peptide antigens before they function as antigen-presenting cells. This preliminary action enhances their cytotoxic activity and ability to present antigens, ensuring that when these cells are used to induce CTLs, both cell types achieve therapeutic-level functionality and numbers from small blood volumes.
Data Source
AI summary
Disclosed are: a method for culture of disease antigen specific CTLs and γδT cells in one culture step conveniently and efficiently; and a pharmaceutical agent and a therapeutic/prophylactic method both of which use a cell produced by the method. Blood is collected and peripheral blood mononuclear cells are separated from the blood. Aminobisphosphonate and a disease antigen are added to the peripheral blood mononuclear cells at the beginning of culture, and the cell culture is carried out for a predetermined period to proliferate/induce disease antigen specific CTLs and γδT cells simultaneously until the numbers of the cells reach values that are effective for the treatment of a disease. The CTLs and the γδT cells thus produced are used for the treatment.