Abstract
BackgroundReactivation of latent viruses such as human cytomegalovirus (HCMV) after allogeneic hematopoietic stem cell transplantation (HSCT) results in high morbidity and mortality. Effective immunization against HCMV shortly after allo-HSCT is an unmet clinical need due to delayed adaptive T cell development. Donor-derived dendritic cells (DCs) have a critical participation in stimulation of naïve T cells and immune reconstitution, and therefore adoptive DC therapy could be used to protect patients after HSCT. However, previous methods for ex vivo generation of adoptive donor-derived DCs were complex and inconsistent, particularly regarding cell viability and potency after thawing. We have previously demonstrated in humanized mouse models of HSCT the proof-of-concept of a novel modality of lentivirus-induced DCs (“SmyleDCpp65”) that accelerated antigen-specific T cell development.MethodsHere we demonstrate the feasibility of good manufacturing practices (GMP) for production of donor-derived DCs consisting of monocytes from peripheral blood transduced with an integrase-defective lentiviral vector (IDLV, co-expressing GM-CSF, IFN-α and the cytomegalovirus antigen pp65) that were cryopreserved and thawed.ResultsUpscaling and standardized production of one lot of IDLV and three lots of SmyleDCpp65 under GMP-compliant conditions were feasible. Analytical parameters for quality control of SmyleDCpp65 identity after thawing and potency after culture were defined. Cell recovery, uniformity, efficacy of gene transfer, purity and viability were high and consistent. SmyleDCpp65 showed only residual and polyclonal IDLV integration, unbiased to proto-oncogenic hot-spots. Stimulation of autologous T cells by GMP-grade SmyleDCpp65 was validated.ConclusionThese results underscore further developments of this individualized donor-derived cell vaccine to accelerate immune reconstitution against HCMV after HSCT in clinical trials.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-015-0599-5) contains supplementary material, which is available to authorized users.
Highlights
Reactivation of latent viruses such as human cytomegalovirus (HCMV) after allogeneic hematopoietic stem cell transplantation (HSCT) results in high morbidity and mortality
Advancing towards clinical grade SmyleDCpp65 for clinical trials, we evaluated the feasibility of vector production and cell generation under good manufacturing practices (GMP) compliant conditions
GMP-grade Integrase-defekten lentiviralen Vektor (IDLV)-G2α2pp65 production and recovery demonstrated that IDLV production was not fundamentally different from ICLV production methods established by the same CMO
Summary
Reactivation of latent viruses such as human cytomegalovirus (HCMV) after allogeneic hematopoietic stem cell transplantation (HSCT) results in high morbidity and mortality. Effective immunization against HCMV shortly after allo-HSCT is an unmet clinical need due to delayed adaptive T cell development. Donor-derived dendritic cells (DCs) have a critical participation in stimulation of naïve T cells and immune reconstitution, and adoptive DC therapy could be used to protect patients after HSCT. Cytoreductive conditioning regimens, T cell depletion and immune suppressive therapies used in the context of allo-HSCT elicit a delay in adaptive immunity, predisposing patients to infections. Among infections and reactivations after allo-HSCT, human cytomegalovirus (HCMV) is a major challenge for clinicians and patients due to high morbidity, mortality and significant costs for management with antiviral drugs or adaptive T cell therapy [3, 4]. Simple innovative, relatively inexpensive and individualized cell therapy approaches are warranted to cover this unmet clinical need
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