CMV, EBV and adenovirus are problematic in patients after stem cell (SCT) and cord blood transplantation (CBT) and are associated with morbidity and mortality. Deficiencies in conventional therapeutics have increased interest in an immunotherapeutic approach to viral disorders. We have developed 2 strategies to grow multivirus-specific donor-derived T-cells (mCTL), one from peripheral blood (PB) of adult CMV-seropositive donors and another from naive cord blood (CB). Using an adenoviral-vector expressing CMVpp65 or overlapping viral peptides for CMV (pp65 and IE-1), EBV (EBNA1 and LMP2), and Adenovirus (Hexon and Penton) presented to T cells by dendritic cells, monocytes, or EBV-LCL, we generated a single culture of mCTL. PB mCTL (Mean SFC:adeno:666, EBV:129, CMV:535) had more spot forming cells (SFC per 100,000 cells) (Mean, adeno: 666, EBV: 129, CMV: 535) than CB mCTL (adeno:117, EBV:95, CMV:67) by IFN-gamma ELISPOT assay but both contained cells specific for at least 1 virus. mCTL derived from both CB and PB contained a mixture of CD4+ and CD8+ T cells with an effector and central memory phenotype. Based on deep T cell receptor sequencing, CB mCTL were more polyclonal than PB-derived mCTL.
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