Manufacturing synthetic viscoelastic antigen-presenting cells for immunotherapy.
Publication Year:
2026
PubMed ID:
41068478
Funding Grants:
Public Summary:
The immune system plays an important role in protecting the body against diseases, including cancer. T cells are a type of immune cell that can recognize and destroy cancer cells. However, using T cells effectively for cancer treatment requires activating and growing large numbers of these cells outside the body. Current methods often rely on artificial materials that do not fully reproduce the natural environment in which T cells become activated, potentially limiting their effectiveness. To address this challenge, we developed a method to create tiny, soft, artificial cells that mimic the natural cells responsible for activating T cells in the body. These artificial cells are made from alginate, a safe, gel-like material derived from seaweed. Unlike conventional hard particles, our artificial cells can be designed to have different levels of softness, flexibility, and the ability to gradually change shape under pressure. These physical properties help them better resemble natural cells and interact more effectively with T cells. Using a specialized technology that produces tiny droplets, we can manufacture large numbers of these artificial cells with consistent sizes and carefully controlled properties. We then attach specific molecules to their surfaces that provide the signals needed to activate T cells. Our studies show that these artificial cells can effectively activate T cells and support their growth. They also encourage the production of T cells with long-lasting immune memory, which may help provide more sustained protection against cancer. In addition, the technology improves the preparation of genetically engineered T cells, including those used in CAR-T cell therapy, a promising cancer treatment that modifies a patient's immune cells to recognize and attack cancer. In laboratory and animal studies, T cells prepared using these artificial cells demonstrated improved cancer-killing ability. An additional advantage is that the artificial cells can be easily separated from the activated T cells after use, helping preserve the cells intended for treatment. The entire preparation and testing process takes approximately one week. This protocol provides researchers with a practical and adaptable approach to producing artificial cells for T cell activation. By more closely recreating the natural conditions needed for immune cell activation, this technology could contribute to the development of more effective and potentially longer-lasting cancer immunotherapies.
Scientific Abstract:
This protocol details the preparation and functionalization of viscoelastic synthetic antigen-presenting cells (APCs) for T cell activation, designed to enhance immunotherapeutic efficacy. Using a high-throughput microfluidic system and post-processing, we create cell-sized sodium alginate microbeads with tunable stiffness, viscoelasticity and surface chemistry, enabling them to better mimic the physical and activation properties of natural APCs. The protocol includes fabrication of synthetic cells with defined sizes, crosslinking strategies to achieve desirable mechanical properties, surface functionalization via click chemistry for attaching activation molecules, and characterization methods for mechanical and biochemical properties. Compared with traditional matrices or rigid microbeads, this approach allows precise control over the mechanical and biochemical features of synthetic APCs, ensuring optimal T cell activation. The resulting synthetic cells support robust T cell activation and expansion, enhance the CD8/CD4 T cell ratio, promote T memory stem cell (TMSC) formation and improve chimeric antigen receptor transduction efficiency, leading to superior tumor-killing efficacy in vitro and in vivo. Additionally, these synthetic cells can be efficiently removed from T cells after activation using simple centrifugation or calcium chelation, preserving the activated T cells. The complete protocol, including fabrication, functionalization and quality assessment, requires ~1 week to complete. Users should have experience in microfluidics, biomaterial handling, bioconjugation techniques and basic cell culture. This platform can be adapted for broader applications in immune cell engineering.