EB病毒特异性T细胞的体外功能研究

Objective This study aimed to establish and validate a rapid manufacturing protocol for EBV-VSTs. Methods Peripheral blood mononuclear cells(PBMCs) of healthy donors were stimulated with EBV antigen peptide pool. Interferon-gamma(IFN-γ) capture assay rapidly isolated and expanded EBV-specific T cells(EBV-VSTs). In vitro functional studies validated the in vitro efficacy and safety of EBV-VSTs. Results The IFN-γ capture assay rapidly enriched EBV-VSTs within CD3+ cell populations up to 91.65%(Range 89.4%～93.5%). Enriched EBV-VSTs exhibited robust antigen-specific IFN-γ secretion(47 021 SFC/106 cells vs. 150 SFC/106 cells, P<0.05), potent proliferative capacity(18 122 cpm vs. 513 cpm, P<0.05), and specific cytotoxicity against target cells(P<0.05). In addition, their alloreactivity was markedly lower than that of unsorted PBMCs(P<0.05). Conclusion The IFN-γ capture-based strategy enables rapid and high-efficiency production of EBV-VSTs with prominent in vitro efficacy and safety profiles. It provides a novel therapeutic strategy for EBV-associated complications post allo-HSCT.