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基于阳离子表面活性剂的黏附型纳米疫苗的新进展,用于红罗非鱼(Oreochromis sp.)中东方气单胞菌亚种的直接浸浴免疫接种。

Novel development of cationic surfactant-based mucoadhesive nanovaccine for direct immersion vaccination against Francisella noatunensis subsp. orientalis in red tilapia (Oreochromis sp.).

机构信息

Center of Excellence in Fish Infectious Diseases (CE FID), Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330, Thailand.

Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330, Thailand.

出版信息

Fish Shellfish Immunol. 2022 Aug;127:1051-1060. doi: 10.1016/j.fsi.2022.07.056. Epub 2022 Jul 21.

Abstract

Francisella noatunensis subsp. orientalis (Fno) is one of the infectious diseases that causes economic losses associated with tilapia mortality. Even though direct immersion administration of vaccines is more practicable for small fish and fry compared with oral and injection vaccination in the fields, the efficacy is still insufficient due to lower potency of antigen uptake. Herein, we accomplished the development of a mucoadhesive nanovaccine platform using cetyltrimethylammonium bromide (CTAB), a cationic surfactant, to improve the efficiency of immersion vaccination against Fno in tilapia. Cationic Fno nanovaccine (CAT-Fno-NV) was prepared though emulsification using an ultrasonic method. In our investigation, the CAT-Fno-NV increased the opportunity of Fno vaccine uptake by extending the contact time between vaccine and mucosal surface of fish gills and enhancing the protective efficacy against Fno infection. Fish were vaccinated with the CAT-Fno-NV by a direct immersion protocol. The challenge trial by Fno injection revealed that CAT-Fno-NV at the concentration 1:100 ratio (approximately 1 × 10 cfu/mL) had the highest efficacy to protect fish from Fno infection at day 30 after post challenge period according to the total number of Fno detected in head kidney, spleen and liver. A significant upregulation of IgM gene was observed in gills, skin, head kidney, serum and peripheral blood lymphocytes (PBLs) and spleen tissues treated with WC and CAT-Fno-NV (1:100) vaccines, while IgT gene was highly expressed in only gills and skin tissues for treated WC and CAT-Fno-NV (1:100) groups. We anticipate that the cationic surfactant-based nanovaccine developed in this study could become an efficient alternative for direct immersion vaccination to induce humoral immune responses against Fno in vaccinated tilapia.

摘要

东方气单胞菌亚种。东方亚种(Fno)是一种传染性疾病,会导致罗非鱼死亡相关的经济损失。尽管与口服和注射疫苗相比,直接浸泡给药在现场更适用于小鱼和鱼苗,但由于抗原摄取效力较低,其功效仍然不足。在此,我们使用阳离子表面活性剂十六烷基三甲基溴化铵(CTAB)完成了一种粘膜粘附纳米疫苗平台的开发,以提高罗非鱼对 Fno 的浸泡免疫效率。阳离子 Fno 纳米疫苗(CAT-Fno-NV)是通过超声乳化法制备的。在我们的研究中,CAT-Fno-NV 通过延长疫苗与鱼鳃粘膜表面的接触时间并增强对 Fno 感染的保护效果,增加了 Fno 疫苗的摄取机会。鱼通过直接浸泡方案接种 CAT-Fno-NV。通过 Fno 注射进行的挑战试验表明,根据攻毒后 30 天检测到头肾、脾脏和肝脏中 Fno 的总数,在 1:100 浓度比(约 1×10cfu/mL)下的 CAT-Fno-NV 具有最高的保护鱼免受 Fno 感染的功效。在用 WC 和 CAT-Fno-NV(1:100)疫苗处理的鱼鳃、皮肤、头肾、血清和外周血淋巴细胞(PBL)和脾脏组织中观察到 IgM 基因的显著上调,而 IgT 基因仅在处理的鱼鳃和皮肤组织中高度表达 WC 和 CAT-Fno-NV(1:100)组。我们预计,本研究开发的基于阳离子表面活性剂的纳米疫苗可以成为直接浸泡免疫的有效替代方法,以诱导接种罗非鱼对 Fno 的体液免疫反应。

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