We recently reported the safety of allogeneic virus-specific T cells (VSTs) expressing a chimeric antigen receptor (CAR) for CD19 (CD19.CAR-VSTs) for the treatment of patients with relapsed B-cell malignancies after HSCT (Cruz et al, Blood 2013). However, the CAR-VSTs did not expand as expected after infusion, even in the presence of viral reactivation. During manufacture, VSTs were cultured ex vivo for 19 days before transduction (late-transduced) with CAR and we hypothesized that by this time, sufficient T-cell differentiation may have occurred limiting their proliferative capacity. To test this hypothesis, we developed and optimized a good manufacturing practices (GMP) compliant method for the early transduction of tri-VSTs directed to Epstein-Barr virus, adenovirus and cytomegalovirus with CAR directed to CD19.
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