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壳聚糖修饰的纳米乳剂在鼻腔疫苗递送中的应用

[Applying Chitosan-Modified Nanoemulsion in Nasal Vaccine Delivery].

作者信息

Hao Xin-Yan, Zhang Yuan-Dong, Hou Ying-Ying, Sun Xun

机构信息

Key Laboratory of Drug-Targeting and Drug Delivery System of the Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2021 Jul;52(4):592-597. doi: 10.12182/20210760104.

Abstract

OBJECTIVE

To prepare a chitosan-modified cationic nanoemulsion that could be used to prolong the residence time of nasal vaccines in the nasal cavity and improve the cellular uptake efficiency so as to enhance the immune efficacy of nasal vaccines.

METHODS

A nanoemulsion-based vaccine coated with chitosan was prepared, and the particle size, potential, antigen encapsulation efficiency, stability as well as cytotoxicity were examined. The uptake efficiency of vaccine on different cells and the residence time of vaccine in the nasal cavity were measured. Finally, nasal vaccine was administered on mice and the antibody levels in the serum and in the nasal lavage fluids of the immunized mice were examined.

RESULTS

The nanoemulsion-based vaccine had an average particle size of (167.2±0.75) nm, a polydispersity index (PDI) of 0.21±0.01, and an average potential of (13.7±0.85) mV. The encapsulation efficiency of antigen was 92.7%. The nanoemulsion-based vaccine had good stability and did not show obvious cytotoxicity in Madin-Darby canine kidney (MDCK) epithelial cells. The vaccine demonstrated relatively high cellular uptake of antigens on DC2.4 and MDCK cells at (49.7±3.45)% and (59.7±2.19)%, respectively. Besides, the cationic nanoemulsion also significantly increased the residence time of the antigen, and a considerable amount of nanoemulsion-based vaccine was found remaining in the nasal cavity 60 minutes after administration. Compared with free antigen and the nanoemulsion without chitosan modification, the chitosan-modified nanoemulsion vaccine induced higher systemic and mucosal antibody levels in mice after nasal immunization ( <0.01).

CONCLUSION

The chitosan-modified nanoemulsion vaccine prepared in the study can enhance the immune efficacy of nasal vaccines, showing great potential to be used as a delivery carrier for nasal vaccines.

摘要

目的

制备一种壳聚糖修饰的阳离子纳米乳剂,用于延长鼻腔疫苗在鼻腔内的停留时间,提高细胞摄取效率,从而增强鼻腔疫苗的免疫效果。

方法

制备壳聚糖包被的纳米乳剂疫苗,并检测其粒径、电位、抗原包封率、稳定性以及细胞毒性。测定疫苗在不同细胞上的摄取效率和在鼻腔内的停留时间。最后,对小鼠接种鼻腔疫苗,检测免疫小鼠血清和鼻腔灌洗液中的抗体水平。

结果

纳米乳剂疫苗的平均粒径为(167.2±0.75)nm,多分散指数(PDI)为0.21±0.01,平均电位为(13.7±0.85)mV。抗原包封率为92.7%。纳米乳剂疫苗具有良好的稳定性,在犬肾上皮细胞(MDCK)中未表现出明显的细胞毒性。该疫苗在DC2.4和MDCK细胞上对抗原的摄取率相对较高,分别为(49.7±3.45)%和(59.7±2.19)%。此外,阳离子纳米乳剂还显著延长了抗原的停留时间,给药60分钟后在鼻腔内发现有相当数量的纳米乳剂疫苗残留。与游离抗原和未修饰壳聚糖的纳米乳剂相比,壳聚糖修饰的纳米乳剂疫苗经鼻腔免疫后在小鼠体内诱导产生了更高的全身和黏膜抗体水平(P<0.01)。

结论

本研究制备的壳聚糖修饰纳米乳剂疫苗可增强鼻腔疫苗的免疫效果,作为鼻腔疫苗的递送载体具有很大潜力。

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