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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