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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...

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Detalles Bibliográficos
Autores principales: Zeng, Qin, Han, Jianfeng, Zhao, Dong, Gong, Tao, Zhang, Zhirong, Sun, Xun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove Medical Press 2012
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.
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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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