Antigen-scaffolds loaded with hyper-stable neoleukin-2/15 expand antigen-specific T cells with a favorable phenotype for adoptive cell therapy.

Introduction: Adoptive cell therapy (ACT) has shown promising results in cancer treatment, however, achieving effective ex vivo expansion of potent, functionally active, and cytotoxic T cells remains challenging. To address this challenge, we recently developed artificial antigen-presenting scaffolds (Ag-scaffolds) capable of expanding antigen-specific T cells with phenotypes favorable for ACT. Here, we compared the established technology using IL2/IL21-loaded Ag-scaffolds (Ag-IL2/21) with scaffolds incorporating Neoleukin-2/15 (Ag-Neo2/15), an engineered cytokine that selectively signals via IL-2Rβ/γ complexes to enhance CD8 Methods: Antigen-specific T cells were expanded ex vivo using Ag-IL2/21 or Ag-Neo2/15 scaffolds. Expansion efficiency, cytokine production, and cytotoxic activity were assessed. Functional and transcriptional states were profiled using single-cell sequencing, including cytotoxic and dysfunction gene signature scoring. T cell receptor (TCR) sequencing was performed to evaluate clonal expansion. Results: Ag-Neo2/15 scaffolds supported robust expansion of antigen-specific CD8 Discussion: Ag-Neo2/15 scaffolds enhance the quality of