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Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus
BACKGROUND: The generation of anti-adenovirus neutralizing antibody (NAb) in humans severely restricts the utilization of recombinant adenovirus serotype 5 (Ad5) vectors in gene therapy for a wide range of clinical trials. To overcome this limitation, we ionically complexed Ad5 with a newly synthesi...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Dove Medical Press
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3299205/ https://www.ncbi.nlm.nih.gov/pubmed/22412299 http://dx.doi.org/10.2147/IJN.S27526 |
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author | Zeng, Qin Han, Jianfeng Zhao, Dong Gong, Tao Zhang, Zhirong Sun, Xun |
author_facet | Zeng, Qin Han, Jianfeng Zhao, Dong Gong, Tao Zhang, Zhirong Sun, Xun |
author_sort | Zeng, Qin |
collection | PubMed |
description | BACKGROUND: The generation of anti-adenovirus neutralizing antibody (NAb) in humans severely restricts the utilization of recombinant adenovirus serotype 5 (Ad5) vectors in gene therapy for a wide range of clinical trials. To overcome this limitation, we ionically complexed Ad5 with a newly synthesized copolymer, which we called APC, making an adenovirus shielded from NAb. METHODS: APC, a cationic polyethylene glycol derivative, was synthesized via two steps of ring-opening copolymerization of ethylene oxide and allyl glycidyl ether, followed by the addition of 2-mercaptoethylamine. The copolymer or the control PEI-2k was ionically complexed to anionic Ad5 in 5% glucose, and in vitro transduction assays were carried out in coxsackievirus and adenovirus receptor-positive cells (A549) and coxsackievirus and adenovirus receptor-negative cells (B16 and SKOV3). The physical properties and morphology of adenovirus alone or the complexes were investigated respectively by zeta potential, size distribution, and transmission electron microscopy image. Then cytotoxicity of APC was examined using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide assays. Finally, the ability of APC to protect adenovirus from NAb was evaluated by transfection assays after a neutralizing effect. RESULTS: APC was successfully synthesized and showed a low cytotoxicity. Positively charged Ad5/APC exhibited slightly increased diameter (130.2 ± 0.60 nm) than naked Ad5 (115.6 ± 5.46 nm) while Ad5/PEI-2k showed severe aggregation (1382 ± 79.9 nm). Ad5/APC achieved a gene transfection level as high as Ad5/PEI-2k in A549 or B16 cells, and significantly higher than Ad5/PEI-2k in SKOV3 cells. Most importantly, after the exposure to the neutralizing antibody, naked Ad5 and Ad5/PEI-2k exhibited poor gene expression while Ad5/APC still showed significantly efficient gene expression. CONCLUSION: Our results demonstrated that Ad5/APC complex offered good protection for Ad5 against NAb in vitro and suggested a potential strategy of resistance to NAb in vivo. |
format | Online Article Text |
id | pubmed-3299205 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Dove Medical Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-32992052012-03-12 Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus Zeng, Qin Han, Jianfeng Zhao, Dong Gong, Tao Zhang, Zhirong Sun, Xun Int J Nanomedicine Original Research BACKGROUND: The generation of anti-adenovirus neutralizing antibody (NAb) in humans severely restricts the utilization of recombinant adenovirus serotype 5 (Ad5) vectors in gene therapy for a wide range of clinical trials. To overcome this limitation, we ionically complexed Ad5 with a newly synthesized copolymer, which we called APC, making an adenovirus shielded from NAb. METHODS: APC, a cationic polyethylene glycol derivative, was synthesized via two steps of ring-opening copolymerization of ethylene oxide and allyl glycidyl ether, followed by the addition of 2-mercaptoethylamine. The copolymer or the control PEI-2k was ionically complexed to anionic Ad5 in 5% glucose, and in vitro transduction assays were carried out in coxsackievirus and adenovirus receptor-positive cells (A549) and coxsackievirus and adenovirus receptor-negative cells (B16 and SKOV3). The physical properties and morphology of adenovirus alone or the complexes were investigated respectively by zeta potential, size distribution, and transmission electron microscopy image. Then cytotoxicity of APC was examined using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide assays. Finally, the ability of APC to protect adenovirus from NAb was evaluated by transfection assays after a neutralizing effect. RESULTS: APC was successfully synthesized and showed a low cytotoxicity. Positively charged Ad5/APC exhibited slightly increased diameter (130.2 ± 0.60 nm) than naked Ad5 (115.6 ± 5.46 nm) while Ad5/PEI-2k showed severe aggregation (1382 ± 79.9 nm). Ad5/APC achieved a gene transfection level as high as Ad5/PEI-2k in A549 or B16 cells, and significantly higher than Ad5/PEI-2k in SKOV3 cells. Most importantly, after the exposure to the neutralizing antibody, naked Ad5 and Ad5/PEI-2k exhibited poor gene expression while Ad5/APC still showed significantly efficient gene expression. CONCLUSION: Our results demonstrated that Ad5/APC complex offered good protection for Ad5 against NAb in vitro and suggested a potential strategy of resistance to NAb in vivo. Dove Medical Press 2012 2012-02-27 /pmc/articles/PMC3299205/ /pubmed/22412299 http://dx.doi.org/10.2147/IJN.S27526 Text en © 2012 Zeng et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited. |
spellingShingle | Original Research Zeng, Qin Han, Jianfeng Zhao, Dong Gong, Tao Zhang, Zhirong Sun, Xun Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus |
title | Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus |
title_full | Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus |
title_fullStr | Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus |
title_full_unstemmed | Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus |
title_short | Protection of adenovirus from neutralizing antibody by cationic PEG derivative ionically linked to adenovirus |
title_sort | protection of adenovirus from neutralizing antibody by cationic peg derivative ionically linked to adenovirus |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3299205/ https://www.ncbi.nlm.nih.gov/pubmed/22412299 http://dx.doi.org/10.2147/IJN.S27526 |
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