Abstract
The manufacturing process of regulatory T (Treg) cells for clinical application begins with the positive selection of CD25+ cells using superparamagnetic iron oxide nanoparticle (SPION)-conjugated anti-CD25 antibodies (spCD25) and immunomagnetic cell separation technology. Our findings revealed that the interaction of spCD25 with its cell target induced the internalization of the complex spCD25–interleukin-2 receptor. Accumulation of intracellular spCD25 triggered oxidative stress, causing delayed Treg expansion and temporary reduction in suppressor activity. This activation delay hindered the efficient generation of clinically competent cells. During this early phase, Treg cells exhibited elevated mitochondrial superoxide and lipid peroxidation levels, with a concomitant decrease in mitochondrial respiration rates. The results uncovered the increased mitochondrial unfolded protein response. This protective, redox-sensitive activity is inherent in Tregs when contrasted with homologous, spCD25-treated, conventional T cells. Although the temporary effects of spCD25 on clinically competent cells did not impede their use in a safety/feasibility pilot study with kidney transplant recipients, it is reasonable to anticipate a potential reduction in their therapeutic efficacy. The mechanistic understanding of the adverse effects triggered by spCD25 is crucial for improving the manufacturing process of clinically competent Treg cells, a pivotal step in the successful implementation of immune cell therapy in transplantation.
Document Type
Article
Publication Date
2024
Digital Object Identifier (DOI)
https://doi.org/10.1016/j.ajt.2024.10.024
Funding Information
This research was supported by the University of Kentucky Alliance Research Initiative (TILT Alliance) and by the National Center for Advancing Translational Sciences, NIH Grant UL1TR001998, to R.G. and F.M. F.C. is supported by the National Institute for Biomedical Imaging and Bioengineering (R03EB035177). The clinical study was partially supported by Novartis. The University of Kentucky Redox Metabolism (RMSR) and the Flow Cytometry and Immune Monitoring (FCIM) shared resource facilities were supported in part by the Office of the Vice President for Research and an NCI Center Core Support Grant (P30 CA177558) to the University of Kentucky Markey Cancer Center.
Repository Citation
Gedaly, Roberto; Orozco, Gabriel; Lewis, Lillie J.; Valvi, Deepa; Chapelin, Fanny; Khurana, Aman; Hidalgo, Giovanna E.; Shmookler, Aaron; Tripathi, Aashutosh; Zhang, Cuiping; Zwischenberger, Joseph B.; and Marti, Francesc, "Effect of mitochondrial oxidative stress on regulatory T cell manufacturing for clinical application in transplantation: Results from a pilot study" (2024). Markey Cancer Center Faculty Publications. 252.
https://uknowledge.uky.edu/markey_facpub/252
Notes/Citation Information
1600-6135/© 2024 The Authors. Published by Elsevier Inc. on behalf of American Society of Transplantation & American Society of Transplant Surgeons. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).