Acosta-Benites Janet, Jara Luis M, Verastegui Pimentel Manuela, Obregón Maldonado Pepe M, Altamirano-Zevallos Faride, Valencia Mamani Nicasio, Gavidia Cesar M
Facultad de Farmacia y Bioquímica, Universidad Nacional Mayor de San Marcos, Lima, Perú.
Facultad de Medicina Veterinaria y Zootecnia, Universidad Peruana Cayetano Heredia, Lima, Perú.
Rev Peru Med Exp Salud Publica. 2025 Jan 31;41(4):411-416. doi: 10.17843/rpmesp.2024.414.13854.
Motivation for the study. Cystic echinococcosis is a neglected disease associated with contact between dogs, humans and sheep. In countries such as Peru, control programs include vaccination of sheep; however, vaccination in dogs is a late control strategy to eliminate the adult parasite or to avoid infection with eggs in the environment. Main findings. We were able to clone and express a recombinant protein (tropomyosin) of the adult parasite in a bacterial system with immunogenic properties. Implications. Obtaining the tropomyosin recombinant protein from E. granulosus allows the development of vaccine candidates in dogs and the exploration of diagnostic tests in hosts. This study aimed to clone, express and produce the recombinant Echinococcus granulosus tropomyosin isoform A protein (EgTrpA) that maintains its antigenic and immunogenic properties as a potential vaccine candidate for dogs and sheep. The Echinococcus granulosus tropomyosin protein (EgTrp) gene was cloned into two vectors: Tropo/His-tag [pET28a (+)] and Tropo/GST-tag (pGEX6P-1). It was then expressed in E. coli BL21. Protein identity was determined by two-dimensional electrophoresis. Immunogenicity and antigenicity were verified by immunizing rabbits with each recombinant protein and assessed by western blot and ELISA. Two-dimensional electrophoresis identified the recombinant EgTrp protein as isoform A. The recombinant proteins showed recognition reactions on Western Blot and serum from immunized rabbits showed an increase in Tropo/His-tag IgG antibodies similar to Tropo/GST-tag. The recombinant EgTrpA protein showed antigenic and immunogenic characteristics in laboratory animals.
研究动机。囊型包虫病是一种与狗、人类和绵羊接触相关的被忽视疾病。在秘鲁等国家,防控项目包括给绵羊接种疫苗;然而,给狗接种疫苗是一种晚期防控策略,用于消除成虫寄生虫或避免在环境中感染虫卵。主要发现。我们能够在具有免疫原性的细菌系统中克隆并表达成虫寄生虫的一种重组蛋白(原肌球蛋白)。意义。从细粒棘球绦虫获得原肌球蛋白重组蛋白有助于开发狗用候选疫苗,并探索宿主中的诊断测试。本研究旨在克隆、表达并生产细粒棘球绦虫原肌球蛋白同工型A蛋白(EgTrpA),该蛋白保持其抗原性和免疫原性,作为狗和绵羊的潜在候选疫苗。细粒棘球绦虫原肌球蛋白蛋白(EgTrp)基因被克隆到两个载体中:Tropo/His标签[pET28a(+)]和Tropo/GST标签(pGEX6P-1)。然后在大肠杆菌BL21中表达。通过二维电泳确定蛋白同一性。通过用每种重组蛋白免疫兔子来验证免疫原性和抗原性,并通过蛋白质印迹法和酶联免疫吸附测定法进行评估。二维电泳将重组EgTrp蛋白鉴定为同工型A。重组蛋白在蛋白质印迹上显示出识别反应,免疫兔子的血清显示Tropo/His标签IgG抗体增加,类似于Tropo/GST标签。重组EgTrpA蛋白在实验动物中显示出抗原性和免疫原性特征。