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日本血吸虫成虫阶段特异性重组蛋白的构建及其鼠抗体反应的评价。

Construction and mouse antibody response evaluation of juvenile stage-specific chimeric protein from Fasciola gigantica.

机构信息

Faculty of Allied Health Sciences and Research unit of vaccine and diagnosis of parasitic diseases, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.

Faculty of Allied Health Sciences and Research unit of vaccine and diagnosis of parasitic diseases, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.

出版信息

Vet Parasitol. 2024 Oct;331:110254. doi: 10.1016/j.vetpar.2024.110254. Epub 2024 Jul 14.

Abstract

Fasciolosis, caused by the liver fluke Fasciola gigantica, is a major parasitic disease that affects livestock and therefore causes significant economic losses in tropical countries. Although anthelminthic drugs can kill the parasite, drug-resistant liver fluke populations are increasing. In this study, a recombinant F. gigantica chimeric protein (rFgCHI) consisting of cathepsin L1H (FgCL1H), cathepsin B3 (FgCB3), and Saposin-like protein 1 (FgSAP1) was designed and expressed in Escherichia coli (BL21). The molecular weight of rFgCHI was 61 kDa. To study the antibody response, male BALB/c mice were immunized via the subcutaneous injection of rFgCHI combined with Quil A. Immunization with rFgCHI showed the induction of IgG1 and IgG2a with a higher IgG1 isotype level, indicating the potential of mixed Th1/Th2 immune responses, with Th2 predominating. However, the results showed high levels of IgG against the single proteins, except for rFgSAP1. Through Western blotting, mouse anti-rFgCHI polyclonal antibodies could be detected to the native proteins obtained from the parasite at all stages. Immunolocalization also revealed that the anti-rFgCHI antibodies could detect targeted antigens in the cecal epithelium of the parasite. These results demonstrated that rFgCHI is immunogenic to the mouse immune system and may potentially be a protein candidate for the development of a fasciolosis vaccine.

摘要

片形吸虫病是由巨型片形吸虫引起的一种主要寄生虫病,它影响家畜,因此给热带国家造成重大经济损失。尽管驱虫药物可以杀死寄生虫,但抗药性的肝片吸虫群体正在增加。在这项研究中,设计并在大肠杆菌 (BL21) 中表达了一种由组织蛋白酶 L1H (FgCL1H)、组织蛋白酶 B3 (FgCB3) 和类 Saposin 蛋白 1 (FgSAP1) 组成的重组巨型片形吸虫嵌合蛋白 (rFgCHI)。rFgCHI 的分子量为 61 kDa。为了研究抗体反应,雄性 BALB/c 小鼠通过皮下注射 rFgCHI 与 Quil A 联合免疫。rFgCHI 免疫诱导产生 IgG1 和 IgG2a,其中 IgG1 同型水平较高,表明存在潜在的混合 Th1/Th2 免疫反应,以 Th2 为主。然而,结果表明,除了 rFgSAP1 之外,针对单一蛋白的 IgG 水平都很高。通过 Western blot,从小鼠抗 rFgCHI 多克隆抗体中可以检测到来自寄生虫各个阶段的天然蛋白。免疫定位还表明,抗 rFgCHI 抗体可以检测到寄生虫盲肠上皮中的靶向抗原。这些结果表明 rFgCHI 对小鼠免疫系统具有免疫原性,可能是开发片形吸虫病疫苗的潜在候选蛋白。

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