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Synthetic DNA Delivery by Electroporation Promotes Robust in Vivo Sulfation of Broadly Neutralizing Anti-HIV Immunoadhesin eCD4-Ig
- Source :
- EBioMedicine
- Publication Year :
- 2018
- Publisher :
- Elsevier BV, 2018.
-
Abstract
- 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.<br />Graphical abstract Unlabelled Image<br />Highlights • Use of DNA/electroporation technology to promote in vivo expression of anti-HIV-1 immunoadhesin eCD4-Ig for 6 months • Engineered a Tyrosylprotein Sulfotransferase 2 variant (IgE-TPST2) that efficiently sulfate eCD4-Ig in vivo at low dose • IgE-TPST2 mediated sulfation enhanced potency of eCD4-Ig by 3-fold in the neutralizaiton of HIV global panel isolates
- Subjects :
- 0301 basic medicine
Research paper
Immunoadhesin
HIV Antibodies
Transfection
General Biochemistry, Genetics and Molecular Biology
Virus
Neutralization
03 medical and health sciences
chemistry.chemical_compound
Plasmid
In vivo
0502 economics and business
Animals
Humans
050207 economics
Mice, Inbred BALB C
050208 finance
biology
Sulfates
Electroporation
05 social sciences
HIV
General Medicine
DNA
Immunoglobulin E
Antibodies, Neutralizing
Cell biology
3. Good health
030104 developmental biology
HEK293 Cells
chemistry
biology.protein
Female
Post-translational modification
Antibody
Enzyme trafficking
Ex vivo
CD4 Immunoadhesins
Subcellular Fractions
Subjects
Details
- ISSN :
- 15565068
- Database :
- OpenAIRE
- Journal :
- SSRN Electronic Journal
- Accession number :
- edsair.doi.dedup.....23e5affd877ab852d80d345a0f67b0c5