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奶牛乳房炎 waaF 亚基构建及其 DNA 疫苗的研制与免疫原性初步研究

Construction of the waaF Subunit and DNA Vaccine Against in Cow Mastitis and Preliminary Study on Their Immunogenicity.

作者信息

Wang Hua, Yuan Ligang, Wang Tao, Cao Lu, Liu Fukang, Song Juanjuan, Zhang Yong

机构信息

Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.

出版信息

Front Vet Sci. 2022 May 12;9:877685. doi: 10.3389/fvets.2022.877685. eCollection 2022.

Abstract

() is one of the major pathogenic bacteria in bovine mastitis, which usually triggers systemic symptoms by releasing lipopolysaccharide (LPS). waaF is the core in LPS pathogenicity. In this study, a new waaF vaccine candidate was identified, constructed with the pcDNA3.1 (+)HisB-waaF plasmid to create to a DNA vaccine (pcwaaF), and transfected into MCF-7 cells to produce recombinant waaF subunit vaccine (rwaaF). After that, the safety of the two vaccine candidates was evaluated in mouse model. Immunogenicity and mortality of challenged mice were compared in 20 and 40 μg per dose, respectively. The results showed that rwaaF and pcwaaF were successfully constructed and the complete blood count and serum biochemical indicated that both of the vaccine candidates were safe ( > 0.05). In addition, histopathological staining showed no obvious pathological changes. The immune response induced by rwaaF was significantly higher than that of pcwaaF ( < 0.01), indicated by levels of serum concentration of IgG IL-2, IL-4, and IFN-γ, and feces concentration of sIgA. Survival rates of mice in rwaaF groups (both 80%) were also higher than in the pcwaaF groups (40 and 50%, respectively). Comparing the safety, immunogenicity, and challenge of two vaccine candidates, rwaaF had the better effect and 20 μg rwaaF was more economical. In conclusion, this study demonstrates the utility of a new vaccine and provides a rationale for further investigation of bovine mastitis therapy and management.

摘要

()是牛乳腺炎的主要致病菌之一,它通常通过释放脂多糖(LPS)引发全身症状。waaF是LPS致病性的核心。在本研究中,鉴定出一种新的waaF候选疫苗,用pcDNA3.1(+)HisB - waaF质粒构建成DNA疫苗(pcwaaF),并转染到MCF - 7细胞中以生产重组waaF亚单位疫苗(rwaaF)。之后,在小鼠模型中评估了这两种候选疫苗的安全性。分别比较了每剂量20μg和40μg时受攻击小鼠的免疫原性和死亡率。结果表明,rwaaF和pcwaaF成功构建,全血细胞计数和血清生化指标表明两种候选疫苗均安全(>0.05)。此外,组织病理学染色未显示明显病理变化。rwaaF诱导的免疫反应明显高于pcwaaF(<0.01),这通过血清中IgG、IL - 2、IL - 4和IFN - γ的浓度水平以及粪便中sIgA的浓度来表明。rwaaF组小鼠的存活率(均为80%)也高于pcwaaF组(分别为40%和50%)。比较两种候选疫苗的安全性、免疫原性和挑战性,rwaaF效果更好,20μg rwaaF更经济。总之,本研究证明了一种新疫苗的实用性,并为进一步研究牛乳腺炎的治疗和管理提供了理论依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23e0/9134013/3f4f6ddd7dca/fvets-09-877685-g0001.jpg

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