In situ T-cell transfection by anti-CD3-conjugated lipid nanoparticles leads to T-cell activation, migration, and phenotypic shift.

Autor: Kheirolomoom A; Stanford University, Department of Radiology, Palo Alto, CA, USA., Kare AJ; Stanford University, Department of Bioengineering, Stanford, CA, USA., Ingham ES; University of California, Davis, Department of Biomedical Engineering, Davis, CA, 95616, USA., Paulmurugan R; Stanford University, Department of Radiology, Palo Alto, CA, USA., Robinson ER; Stanford University, Department of Radiology, Palo Alto, CA, USA., Baikoghli M; University of California, Davis, Department of Molecular and Cellular Biology, Davis, CA, USA., Inayathullah M; Stanford University, Department of Radiology, Palo Alto, CA, USA., Seo JW; Stanford University, Department of Radiology, Palo Alto, CA, USA., Wang J; Stanford University, Department of Radiology, Palo Alto, CA, USA., Fite BZ; Stanford University, Department of Radiology, Palo Alto, CA, USA., Wu B; Stanford University, Department of Radiology, Palo Alto, CA, USA., Tumbale SK; Stanford University, Department of Radiology, Palo Alto, CA, USA., Raie MN; Stanford University, Department of Radiology, Palo Alto, CA, USA., Cheng RH; University of California, Davis, Department of Molecular and Cellular Biology, Davis, CA, USA., Nichols L; Stanford Shared FACS Facility, Stanford University, Stanford, CA, USA., Borowsky AD; University of California, Davis, Center for Comparative Medicine, Davis, CA, USA., Ferrara KW; Stanford University, Department of Radiology, Palo Alto, CA, USA. Electronic address: kwferrar@stanford.edu.
Jazyk: angličtina
Zdroj: Biomaterials [Biomaterials] 2022 Feb; Vol. 281, pp. 121339. Date of Electronic Publication: 2021 Dec 29.
DOI: 10.1016/j.biomaterials.2021.121339
Abstrakt: Ex vivo programming of T cells can be efficacious but is complex and expensive; therefore, the development of methods to transfect T cells in situ is important. We developed and optimized anti-CD3-targeted lipid nanoparticles (aCD3-LNPs) to deliver tightly packed, reporter gene mRNA specifically to T cells. In vitro, targeted LNPs efficiently delivered mCherry mRNA to Jurkat T cells, and T-cell activation and depletion were associated with aCD3 antibody coating on the surface of LNPs. aCD3-LNPs, but not non-targeted LNPs, accumulated within the spleen following systemic injection, with mCherry and Fluc signals visible within 30 min after injection. At 24 h after aCD3-LNP injection, 2-4% of all splenic T cells and 2-7% of all circulating T cells expressed mCherry, and this was dependent on aCD3 coating density. Targeting and transfection were accompanied by systemic CD25 + , OX40 + , and CD69 + T-cell activation with temporary CD3e ligand loss and depletion of splenic and circulating subsets. Migration of splenic CD8a + T cells from the white-pulp to red-pulp, and differentiation from naïve to memory and effector phenotypes, followed upon aCD3-LNP delivery. Additionally, aCD3-LNP injection stimulated the secretion of myeloid-derived chemokines and T-helper cytokines into plasma. Lastly, we administered aCD3-LNPs to tumor bearing mice and found that transfected T cells localized within tumors and tumor-draining lymph nodes following immunotherapy treatment. In summary, we show that CD3-targeted transfection is feasible, yet associated with complex immunological consequences that must be further studied for potential therapeutic applications.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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