Abstract
Background aims
Regulatory (or “tolerogenic”) dendritic cells (DCregs) are a highly promising, innovative
cell therapy for the induction or restoration of antigen-specific tolerance in immune-mediated
inflammatory disorders. These conditions include organ allograft rejection, graft-versus-host
disease following bone marrow transplantation and various autoimmune disorders. DCregs
generated for adoptive transfer have potential to reduce patients’ dependence on non-specific
immunosuppressive drugs that can induce serious side effects and enhance the risk
of infection and certain types of cancer. Here, our aim was to provide a detailed
account of our experience manufacturing and validating comparatively large numbers
of Good Manufacturing Practice–grade DCregs for systemic (intravenous) infusion into
28 organ (liver) transplant recipients and to discuss factors that influence the satisfaction
of release criteria and attainment of target cell numbers.
Results
DCregs were generated in granulocyte-macrophage colony stimulating factor and interleukin
(IL)-4 from elutriated monocyte fractions isolated from non-mobilized leukapheresis
products of consenting healthy adult prospective liver transplant donors. Vitamin
D3 was added on day 0 and 4 and IL-10 on day 4 during the 7-day culture period. Release
and post-release criteria included cell viability, purity, phenotype, sterility and
functional assessment. The overall conversion rate of monocytes to DCregs was 28 ±
8.2%, with 94 ± 5.1% product viability. The mean cell surface T-cell co-inhibitory
to co-stimulatory molecule (programmed death ligand-1:CD86) mean fluorescence intensity
ratio was 3.9 ± 2.2, and the mean ratio of anti-inflammatory:pro-inflammatory cytokine
product (IL-10:IL-12p70) secreted upon CD40 ligation was 60 ± 63 (median = 40). The
mean total number of DCregs generated from a single leukapheresis product (n = 25
donors) and from two leukapheresis products (n = 3 donors) was 489 ± 223 × 106 (n = 28). The mean total number of DCregs infused was 5.9 ± 2.8 × 106 per kg body weight. DCreg numbers within a target cell range of 2.5–10 × 106/kg were achieved for 25 of 27 (92.6%) of products generated.
Conclusions
High-purity DCregs meeting a range of quality criteria were readily generated from
circulating blood monocytes under Good Manufacturing Practice conditions to meet target
cell numbers for infusion into prospective organ transplant recipients.
Key Words
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Article info
Publication history
Published online: January 12, 2023
Accepted:
November 18,
2022
Received:
July 27,
2022
Publication stage
In Press Corrected ProofIdentification
Copyright
© 2022 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.
