A multidimensional workflow profiling of allogeneic virus-specific T cell therapies reveals potency-linked signatures

Allogeneic virus-specific T cell (VST) therapies offer distinct advantages, including scalability, rapid deployment, and manufacturing consistency, and have demonstrated efficacy in multiple clinical trials. However, identifying VST products with high therapeutic potential remains a major hurdle. Here, we present a multidimensional analytical platform that integrates in vitro and in vivo anti-viral reactivity, T cell receptor (TCR) repertoire analysis, gene expression profiling, immunophenotyping, and functional validation in a humanized mouse model. Epstein-Barr virus (EBV)-specific T cells expanded from HLA-diverse healthy donors consistently enriched for TCRs targeting EBV-encoded antigens. Transcriptomic and high-dimensional flow cytometric analyses revealed a distinct effector-associated signature. Importantly, this integrative approach uncovered correlative biomarkers of T cell potency and effector function, validated in an in vivo model of EBV-driven B cell lymphoma. These findings establish a scalable framework for the characterization of allogeneic T cell products and may inform the development of predictive metrics for in vivo efficacy. Allogeneic T cell therapies could be used in therapeutic applications because of their potential for ‘off-the-shelf’ access and standardised production. Here the authors have developed a multidimensional workflow profiling platform for EBV-specific T cell therapy and show that correlative biomarkers of T cell potency and effector function are associated with therapeutic effectiveness in xenogeneic mouse EBV-LCL models.