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通过减毒鼠伤寒沙门氏菌传递延长和增强 EtMIC3-C-MAR 抗柔嫩艾美耳球虫的保护效力。

Prolonging and enhancing the protective efficacy of the EtMIC3-C-MAR against eimeria tenella through delivered by attenuated salmonella typhimurium.

机构信息

Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Shandong Agricultural University, Taian City, Shandong Province, China.

Shandong Huamutianyuan Agriculture and Animal Husbandry Co., Ltd., 1 Gangxing 3 Road, Jinan, Shandong Province, 250101, China.

出版信息

Vet Parasitol. 2020 Mar;279:109061. doi: 10.1016/j.vetpar.2020.109061. Epub 2020 Feb 24.

Abstract

The microneme adhesive repeats (MAR) of Eimeria tenella microneme protein 3 (EtMIC3) are associated with binding to and invasion of host cells. Adhesion and invasion-related proteins or domains are often strongly immunogenic, immune responses mounted against these factors that play a key role in blocking invasion. In the present study, an oral live vaccine consisting of attenuated Salmonella typhimurium X4550 carrying two MAR domains fragment (St-X4550-MAR) was constructed and its protective efficacies were evaluated. The results showed that St-X4550-MAR was more immunogenic and conferred a higher degree of protection than recombinant MAR polypeptide as reflected by increased body weight, decreased oocyst shedding and lesion scores, increased serum IgG and cecal sIgA antibody production, and increasing levels of interferon-γ and interleukin-10. Thus, MAR domains are highly immunogenic and St-X4550-MAR had moderate activity against E. tenella infection by stimulating humoral, mucosal and cellular immunity. Chickens immunized with our constructed live vaccine provided considerable protections as early as at 10 d post-immunization (ACI: 155.17), and maintained higher protection levels at 20 d post-immunization (ACI: 173.66), and at 30 d post-immunization (ACI: 162.4). While the protective efficacy of chickens immunized with the recombinant MAR peptides showed a decreased trend as the post immunization time prolonging. Thus, using live-attenuated S. typhimurium X4550 as a vaccine expression and delivery system can significantly improve the protective efficacy and duration of protective immunity of MAR of EtMIC3.

摘要

柔嫩艾美耳球虫 Microneme 黏附重复序列 (MAR) 与宿主细胞黏附和入侵有关。黏附和入侵相关的蛋白质或结构域通常具有很强的免疫原性,针对这些在阻止入侵中起关键作用的因素产生的免疫反应可以起到阻断入侵的作用。在本研究中,构建了由减毒鼠伤寒沙门氏菌 X4550 携带两个 MAR 结构域片段 (St-X4550-MAR) 组成的口服活疫苗,并评估了其保护效力。结果表明,St-X4550-MAR 比重组 MAR 多肽更具免疫原性,能提供更高程度的保护,这反映在体重增加、卵囊脱落和病变评分降低、血清 IgG 和盲肠 sIgA 抗体产生增加以及干扰素-γ和白细胞介素-10 水平增加。因此,MAR 结构域具有高度的免疫原性,St-X4550-MAR 通过刺激体液、黏膜和细胞免疫对柔嫩艾美耳球虫感染具有中等活性。用我们构建的活疫苗免疫的鸡在免疫后 10 天 (ACI: 155.17) 就提供了相当大的保护,在免疫后 20 天 (ACI: 173.66) 和 30 天 (ACI: 162.4) 保持更高的保护水平。而用重组 MAR 肽免疫的鸡的保护效力随着免疫后时间的延长呈下降趋势。因此,利用减毒鼠伤寒沙门氏菌 X4550 作为疫苗表达和传递系统可以显著提高 EtMIC3 的 MAR 的保护效力和保护免疫持续时间。

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