Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig.

Autor:	Xu Z; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States; Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States., Wise MC; Inovio Pharmaceuticals, Plymouth Meeting, PA 19422, United States., Choi H; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States., Perales-Puchalt A; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States., Patel A; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States., Tello-Ruiz E; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States., Chu JD; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States., Muthumani K; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States., Weiner DB; Vaccine and Immunotherapy Center, The Wistar Institute, Philadelphia, PA 19104, United States. Electronic address: dweiner@wistar.org.
Jazyk:	angličtina
Zdroj:	EBioMedicine [EBioMedicine] 2018 Sep; Vol. 35, pp. 97-105. Date of Electronic Publication: 2018 Aug 30.
DOI:	10.1016/j.ebiom.2018.08.027
Abstrakt:	Background: Despite vigorous and ongoing efforts, active immunizations have yet to induce broadly neutralizing antibodies (bNAbs) against HIV-1. An alternative approach is to achieve prophylaxis with long-term expression of potent biologic HIV-1 inhibitors with Adeno-associated Virus (AAV), which could however be limited by hosts' humoral and cellular responses. An approach that facilitates in vivo production of these complex molecules independent of viral-vectored delivery will be a major advantage. Methods: We used synthetic DNA and electroporation (DNA/EP) to deliver an anti-HIV-1 immunoadhesin eCD4-Ig in vivo. In addition, we engineered a TPST2 enzyme variant (IgE-TPST2), characterized its intracellular trafficking patterns and determined its ability to post-translationally sulfate eCD4-Ig in vivo. Findings: With a single round of DNA injection, a peak expression level of 80-100μg/mL was observed in mice 14 days post injection (d.p.i). The engineered IgE-TPST2 enzyme trafficked efficiently to the Trans-Golgi Network (TGN). Co-administrating low dose of plasmid IgE-TPST2 with plasmid eCD4-Ig enhanced the potency of eCD4-Ig by three-fold in the ex vivo neutralization assay against the global panel of HIV-1 pseudoviruses. Interpretation: This work provides a proof-of-concept for delivering anti-HIV-1 immunoadhesins by advanced nucleic acid technology and modulating protein functions in vivo with targeted enzyme-mediated post-translational modifications. Funding: This work is supported by NIH IPCAVD Grant U19 Al109646-04, Martin Delaney Collaboration for HIV Cure Research and W.W. Smith Charitable Trust. (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze:	MEDLINE
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