建立一種新型微脂體與聚合物之複合體來做為核酸疫苗與次單位疫苗的共同載體

陳莉沂

doi:10.6842/NCTU.2009.00504

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建立一種新型微脂體與聚合物之複合體來做為核酸疫苗與次單位疫苗的共同載體

Development of a novel liposome-polymer transfection complex as co-delivery system for DNA and subunit vaccine

陳莉沂 , Masters　　Advisor：廖光文　　

英文 DOI： 10.6842/NCTU.2009.00504

微脂體；聚乙烯亞胺；核酸疫苗；次單位疫苗；轉染；大豆沙拉油； liposome ； PEI ； DNA vaccine ； subunit vaccine ； transfection ； soybean oil

Abstract │ Reference (30) │ Altmetrics

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In this study, a novel liposome-polymer transfection complex (LPTC) was developed with inexpensive cost and used as DNA or protein delivery system. LPTC was fabricated via the sonication and composed of soybean oil, polyethylenimine (PEI) and polyethylene glycol (PEG). After preparation of LPTC, the morphology of particles and their physical properties were conducted with TEM, DLS and Zeta-Sizer. The particles of LPTC showed round shape and fuzzy edge around the surface of particle captured by TEM. In addition, the particle sizes of LPTC were in the major range of 212.2 nm to 312.1 nm and the zeta-potential (surface charge) was strongly positive (+38.7 mV). Moreover, we examined the ability of DNA condensation and the protection from DNase I digestion in agarose gel electrophoresis. In the in vitro tests, to clarify the gene delivery efficacy, we evaluated the transfection efficiency of LPTC/DNA complexes in Balb/3T3 cells and the transfection efficiency increased as the charge ratios of LPTC to DNA increasing.  LPTC enhanced the cellular uptake of antigen in mouse macrophage cells and also stimulated TNF-alpha release in naïve mice splenocyte that both showed the potential to be adjuvant in vaccine development.  In vivo studies, using H. pyrori relative Hsp60 and Urease B as antigen model, we observed that vaccination of BALB/c mice with LPTC complexed DNA and protein enhanced humoral immune response. Therefore, these results showed that we have developed a DNA and protein delivery system by liposome-polymer transfection complex with inexpensive cost and successfully applied in the development of DNA and subunit vaccine. The success of this design may provide an economical vaccination alternative for farm animal use.

Reference ( 30 ) 〈TOP〉

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5. Wasungu L, Hoekstra D. Cationic lipids, lipoplexes and intracellular delivery of genes. J Control Release 2006 Nov 28;116(2):255-264.
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