ArticleScientific reports2025
Latent EBV reactivation drives aberrant B-cell proliferation during ex vivo tumor-infiltrating lymphocyte expansion from EBV-negative rectal cancer tumor tissue.
Article in Scientific reports, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Recent studies underscore herpesvirus-associated risks, especially HHV6 and Epstein–Barr virus (EBV), in CAR-T and tumor-infiltrating lymphocyte (TIL) therapies, partly due to ex vivo cell manipulation. EBV-driven B cell outgrowth can occur during TIL expansion, and EBV-transformed B cells may exert immunomodulatory functions, potentially impacting TIL therapy efficacy and safety — areas that remain underexplored. Here, we characterized a spontaneously arising wild-type EBV-transformed B cell line, lcl_burn0214, established during ex vivo culture of a tumor specimen from a rectal cancer patient who was EBV-negative. The lcl_burn0214 cell line has undergone 70 passages and was confirmed as a monoclonal B cell line of patient origin via HLA genotyping, immunophenotyping (CD19, CD20), B cell receptor clonotype analysis. RNA-seq analysis revealed that lcl_burn0214’s gene expression closely resembles that of lymphoblastoid cells (r = 0.88) rather than malignant B lymphomas. In immunocompromised mice, lcl_burn0214 exhibited limited tumorigenicity. Autologous TILs were negative for B cells and exhibited a strong interferon-gamma response in the ELISPOT assay during coculture with lcl_burn0214 or with the EBV peptide pool (1000–1800 spots per million of TILs). Our findings underscore the importance of monitoring TIL products for EBV-positive B cell contamination during manufacturing to ensure safety and therapeutic efficacy even for EBV-negative patients.
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