SITC

A single programmable RBC platform supports feeder-free T-cell activation and antigen-specific CAR-T cell expansion, with quantitatively controlled antigen presentation. sbPCI CD3/CD28 produced more T cells at electroporation and more CAR+ cell than competitive nanomatrix. sbPCI CD19/28/137 gave higher CAR+ expansion and moved toward a more balanced CD4:CD8.

Integrated cytokines in this formulation support expansion with reduced metabolic demand while favoring memory associated T cell phenotypes. Synapse-localized cytokine presentation, through surface integration via sbPCI, supported effective T cell expansion in 6M G-Rex while favorably shifting differentiation toward memory-associated, cytotoxically armed phenotypes.

Immune cells respond not only to what signals they receive and how much they receive, but also to how those signals are spatially organized. Signal arrangement influences immune cell phenotype and function independent of receptor identity and signal strength. They product distinct T cell and NK cell responses when presented in different spatial configurations. This indicates that signal geometry represents a programmable and previously underexplored determinant of immune cell outcome.

High-throughput screening of sbPCI configurations reveals that cytokine incorporation collapses donor response heterogeneity from three qualitative expansion phenotypes to a single responsive profile, as visualized by PhenoGraph analysis.

EV-CMS, based on sbPCI technology, has potential to enable more accurate EV detection and control. sbPCI enables quantitative labeling of lipid bilayers, including extracellular vesicles. The incorporation of stoichiometrically-controlled labels that can be detected across distinct modalities enable cross-platform quantitation of extracellular vesicles, with the same EV-CMS.

sbPCI Eliminates Isolation Steps and Enables Rapid Expansion of Rare TCR-specific Cells to Clinically Sufficient Numbers in 8 Days. The technology can be successfully customized to product clinically-relevant numbers of TCR-specific T cells from a large polyclonal population within 8 days. Using sbPCI, CMV TCR-specific CD8+ T cells were selectively expanded from a starting population with low CMV-TCR expression levels. After 48 hours, the expanded T cells produced cytolytic and pro-inflammatory cytokines.

sbPCI enables high-throughput, cost-effective identification of donor-specific formulations for T-cell expansion that outperform industry standards. The technology simplified and accelerated screening with assay setup reduced to 2-4 hours and scale increased to >500 samples/assay. The assay identified CD3, CD28 and CD137 configurations combined with IL-2 that better enabled T cell expansion compared to industry standard. Phenotype-specific formulations were also identified.

Dexamethasone resistant cell therapy is achievable and scalable for patients undergoing long-term immunosuppressive treatment. The treated cells expressed similar levels of cytolytic enzymes as untreated cultures and expanded a majority of CD3+CD8+ cells. The results support the viability of manufacturing potent corticosteroid-resistant therapies for patients.