Recombinant African swine fever virus p30–flagellin fusion protein promotes p30-specific humoral and cellular immune responses in mice

IF 1.4 3区农林科学 Q4 IMMUNOLOGY

Veterinary immunology and immunopathology Pub Date : 2025-01-01 DOI:10.1016/j.vetimm.2024.110864

Xia Huang , Xilong Kang , Shunzi Han , Chuang Meng , Hongqin Song , Xinan Jiao , Zhiming Pan

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Abstract

African swine fever (ASF) is a transmissible and deadly viral disease caused by the African swine fever virus (ASFV) that has considerably affected the global pig industry. Vaccination is considered a potentially effective method to control ASF. However, live attenuated vaccines can't protect against all circulating virus isolates. Subunit vaccines can induce both cellular and humoral immune responses, but often require the addition of adjuvants. Flagellin, a stimulator of Toll-like receptor 5 (TLR5), functions as a potent adjuvant by enhancing cellular and humoral immune responses. However, its high antigenicity may cause severe systemic inflammation. In this study, an Escherichia coli expression system was used to express ASFV p30 protein (p30) fused with Salmonella Typhimurium FliCΔD2D3 (without the D2 and D3 domains of FliC). The immunological effect of p30-FlicΔD2D3 protein in mice was evaluated. Results revealed that the ASFV p30 protein and the p30-FlicΔD2D3 fusion protein were effectively expressed by the E. coli expression system. In vitro activity analysis showed that the p30-FlicΔD2D3 fusion protein could be recognized by ASFV-positive serum, had good immunoreactivity, and remarkably promoted IL-8 secretion related to TLR5 activity in HEK293-mTLR5 cells. However, p30-FlicΔD2D3 induced significantly lower levels of inflammatory factor IL-8 than that induced by wild-type flagellin. Immunization with the p30-FlicΔD2D3 fusion protein considerably promoted cellular and humoral immune responses in mice. Therefore, the p30-FlicΔD2D3 protein retained good immune reactivity and TLR5 agonist efficacy. It also enhanced humoral and cellular immune responses in mice. This work offered valuable information that will be helpful to develop ASF subunit vaccines.

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重组非洲猪瘟病毒 p30-鞭毛蛋白融合蛋白可促进小鼠的 p30 特异性体液和细胞免疫反应。

非洲猪瘟（ASF）是由非洲猪瘟病毒（ASFV）引起的一种传染性和致命的病毒性疾病，严重影响了全球养猪业。疫苗接种被认为是控制非洲猪瘟的潜在有效方法。然而，减毒活疫苗不能预防所有的流行病毒分离株。亚单位疫苗可以诱导细胞和体液免疫反应，但通常需要添加佐剂。鞭毛蛋白是toll样受体5 （TLR5）的刺激剂，作为一种有效的佐剂，通过增强细胞和体液免疫应答而发挥作用。然而，它的高抗原性可能引起严重的全身炎症。本研究利用大肠杆菌表达系统表达ASFV p30蛋白（p30）与鼠伤寒沙门氏菌FliCΔD2D3融合（不含FliC的D2和D3结构域）。评价p30-FlicΔD2D3蛋白对小鼠的免疫作用。结果表明，ASFV p30蛋白和p30-FlicΔD2D3融合蛋白在大肠杆菌表达系统中得到了有效表达。体外活性分析表明p30-FlicΔD2D3融合蛋白能被asfv阳性血清识别，具有良好的免疫反应性，并能显著促进HEK293-mTLR5细胞中与TLR5活性相关的IL-8分泌。然而，p30-FlicΔD2D3诱导的炎症因子IL-8水平明显低于野生型鞭毛蛋白。p30-FlicΔD2D3融合蛋白免疫可显著促进小鼠的细胞和体液免疫反应。因此，p30-FlicΔD2D3蛋白保持了良好的免疫反应性和TLR5激动作用。它还增强了小鼠的体液和细胞免疫反应。这项工作为开发非洲猪瘟亚单位疫苗提供了有价值的信息。

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来源期刊

Veterinary immunology and immunopathology 农林科学-免疫学

CiteScore

3.40

自引率

5.60%

发文量

审稿时长

70 days

期刊介绍： The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease. Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above. The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.