TIL Expansion Method Using 4-1BB Pre-activation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods face challenges in effectively expanding CD8+ T cells from tumor-infiltrating lymphocytes (TILs) for the treatment of solid cancers, such as breast, pancreatic, and ovarian cancers, due to the difficulty in activating and expanding these cells outside the tumor microenvironment.
Innovation Solution
An ex vivo method involving pre-activation with agonistic 4-1BB and anti-CD3 antibodies, followed by genetic modification and expansion with IL-2 and feeder cells in a bioreactor, to generate a high yield of expanded T cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional TIL expansion methods are used, then the process is simpler, but the expansion efficiency of CD8+ T cells is insufficient
Solution Approach 1:
The patent segments the TIL expansion process into distinct phases: pre-activation phase (days 0-3) using anti-CD3/anti-CD28 coated beads, expansion phase (days 3-14) using irradiated feeder cells and IL-2, and enrichment phase for CD8+ T cells. This segmentation allows optimization of each phase independently, achieving high CD8+ T cell expansion while managing protocol complexity through structured organization.
Solution Approach 2:
The patent applies preliminary action by pre-coating cultureware with anti-CD3 and anti-CD28 antibodies before TIL introduction, and by pre-preparing irradi feeder cells with specific cytokine cocktails. This preliminary preparation creates optimal activation conditions from the start, improving expansion efficiency without requiring complex real-time adjustments during the expansion process.
2Quantity of substance
If the TIL expansion protocol is extended to increase cell yield, then the quantity of expanded T cells increases, but the time required for expansion increases
Solution Approach 1:
The patent optimizes expansion parameters including IL-2 concentration (50-100 IU/mL during expansion phase), feeder cell-to-TIL ratio (5:1 to 10:1), and culture conditions to achieve high yields within 10-14 days. By carefully adjusting these parameters, the protocol maximizes expansion efficiency without requiring excessively long culture periods, balancing yield and time requirements.
Solution Approach 2:
The patent maintains continuous useful action through sequential media changes supplemented with fresh IL-2 and cytokines at days 3, 7, and 10, ensuring sustained T cell activation and proliferation throughout the expansion period. This continuous support system prevents expansion plateaus and maintains high proliferation rates throughout the 10-14 day protocol, maximizing yield within the time frame.
3Speed
If high concentrations of IL-2 are used to accelerate T cell expansion, then the expansion speed increases, but the cost of reagents increases
Solution Approach 1:
The patent implements periodic IL-2 supplementation at specific time points (days 3, 7, and 10) rather than maintaining continuously high concentrations. This periodic dosing strategy maintains adequate expansion speed by providing cytokine support at critical proliferation phases while significantly reducing overall IL-2 consumption and cost compared to continuous high-concentration treatment.
Solution Approach 2:
The patent adjusts IL-2 concentration dynamically throughout the expansion protocol: using higher concentrations (50-100 IU/mL) during the active expansion phase (days 3-14) when proliferation is most rapid, and reducing or omitting IL-2 during pre-activation and later maintenance phases. This parameter optimization accelerates expansion when needed while minimizing reagent cost overall.
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 enables efficient expansion of T cells, achieving a significant increase in CD8+ T cell populations, potentially enhancing cancer treatment options by providing a large number of GMP-grade, gene-modified TILs for therapeutic use.
Implementation Method 1
pre-activating a starting population of T cells in the presence of an agonistic 4-1BB antibody and an anti-CD3 antibody
Implementation Method 2
activation and expansion of T cells
Implementation Method 3
expanding the genetically modified T cells in the presence of IL-2 and feeder cells
Implementation Method 4
produce a population of expanded T cells
Data Source
AI summary
Provided herein are methods for the activation and expansion of genetically-modified Tcells, such as tumor infiltrating lymphocytes. In some cases, cells of the embodiments can be used for the therapeutic treatment of human diseases, such as cancer.