Improved expression of recombinant fusion defensin geneplasmids packed with chitosan-derived nanoparticlesand effect on antibacteria and mouse immunity

Wan,Xiaoping; Chen,Jianlin; Cheng,Chi; Zhang,Huabing; Zhao,Shiji; Li,Jianglin; Lv,Xuebin; Wang,Zezhou; Gao,Rong

doi:10.3892/etm.2018.6716

Improved expression of recombinant fusion defensin geneplasmids packed with chitosan-derived nanoparticlesand effect on antibacteria and mouse immunity

Authors:
- Xiaoping Wan
- Jianlin Chen
- Chi Cheng
- Huabing Zhang
- Shiji Zhao
- Jianglin Li
- Xuebin Lv
- Zezhou Wang
- Rong Gao
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Affiliations: Key Laboratory of Bio-Resource and Eco-Environment, Ministry of Education, Key Laboratory of Animal Disease Prevention and Food Safety of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan 610065, P.R. China, School of Laboratory Medicine, Chengdu Medical College, Chengdu, Sichuan 610500, P.R. China, College of Bioengineering, Sichuan University of Science & Engineering, Zigong, Sichuan 643000, P.R. China, Sichuan Academy of Animal Science, Chengdu, Sichuan 610066, P.R. China, Center for Animal Disease Control of Sichuan Province, Chengdu, Sichuan 610035, P.R. China
Published online on: September 11, 2018 https://doi.org/10.3892/etm.2018.6716
Pages: 3965-3972

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Abstract

In order to develop a secure and competent technique to express the human immune gene for fighting infections, we cloned and expressed the BD2/3 using VR1020 (a eukaryotic expression plasmid). BD2/3 contains human β-defensin 2 (BD2) and human BD3. To explore safe and effective DNA delivery molecules in vitro and in vivo, the fusion genes of BD2/3 were used as an immune-labelled gene to verify transfection effectivness of modified chitosan (CS). Plasmid of VR1020-BD2/3 was packed with biomaterials: CS, average molecular weight: 25000D; polyethylene glycol-O-chitosan-polyethylenimine (PEG-O-CS-PEI); liposomes (LP); polyamine cationic liposomes (PCL); polyamine cationic liposomes of protamine (PCL-protamine) by ionotropic gelation. We observed that BD2/3 fusion gene showed high bioactivity in vitro and in vivo. The BD2/3 fusion protein inhibited the proliferation of bacteria (S. aureus, S. pneumoniae, P. aeruginosa and E. coli). The Kunming mice were immune to these nanoparticles and we analyzed their delivery efficiency and gene expression effect. BD2/3 results in multiple changes of innate and required immune system of mice. BD2/3 increases expression of IgG, IgG1, IgG2a, IL-2, IL-6, IFN-γ, as well as of lymphocytes and monocytes. Following challenge with virulent E. coli, CD4+ and CD8+ positive T‑cell counts were highly elevated in the BD2/3 immunized mice, resulting in higher survival rates of mice. These results indicate that nanoparticles containing modified CS and BD2/3 are potentially safe and effective drugs in vivo to improve the immunity against bacterial infection and enhance innate immunity and adaptive immunity against infectious diseases.

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