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旋毛形线虫 Tc24 抗原由裂殖壶菌表达并经口服给药具有免疫原性。

Trypanosoma cruzi Tc24 Antigen Expressed and Orally Delivered by Schizochytrium sp. Microalga is Immunogenic in Mice.

机构信息

Immunology & Vaccinology Group, Centro de Investigaciones Biológicas del Noroeste, S.C., La Paz, BCS, Mexico.

CONACYT-Centro de Investigaciones Biológicas del Noroeste, S.C. (CIBNOR), Col. Playa Palo de Santa Rita Sur, Av. Instituto Politécnico Nacional 195, CP. 23096, La Paz, BCS, Mexico.

出版信息

Mol Biotechnol. 2024 Jun;66(6):1376-1388. doi: 10.1007/s12033-023-00763-6. Epub 2023 Jun 21.

Abstract

Chagas disease-caused by the parasite Trypanosoma cruzi-is a neglected tropical disease for which available drugs are not fully effective in the chronic stage and a vaccine is not available yet. Microalgae represent a promising platform for the production and oral delivery of low-cost vaccines. Herein, we report a vaccine prototype against T. cruzi produced in a microalgae platform, based on the candidate antigen Tc24 with a C terminus fusion with the Co1 peptide (Tc24:Co1 vaccine prototype). After modeling the tertiary structure, in silico studies suggested that the chimeric protein is antigenic, not allergenic, and molecular docking indicated binding with Toll-like receptors 2 and 4. Thus, Tc24:Co1 was expressed in the marine microalga Schizochytrium sp., and Western blot confirmed the expression at 48 h after induction, with a yield of 632 µg/L of algal culture (300 μg/g of lyophilized algal cells) as measured by the enzyme-linked immunosorbent assay (ELISA). Upon oral administration of whole-cell Schizochytrium sp. expressing Tc24:Co1 (7.5 µg or 15 µg of Tc24:Co1 doses) in mice, specific serum IgG and intestinal mucosa IgA responses were detected in addition to an increase in serum Th1/Th2 cytokines. In conclusion, Schizochytrium sp.-expressing Tc24:Co1 is a promising oral vaccine prototype to be evaluated in an animal model of Trypanosoma cruzi infection.

摘要

克氏锥虫病是一种由寄生虫克氏锥虫引起的被忽视的热带病,现有的药物在慢性阶段不能完全有效,而且还没有疫苗。微藻为生产和口服递送低成本疫苗提供了一个有前途的平台。在此,我们报告了一种基于候选抗原 Tc24 与 C 末端融合 Co1 肽的微藻平台生产的针对 T. cruzi 的疫苗原型(Tc24:Co1 疫苗原型)。在对三级结构进行建模后,计算机模拟研究表明该嵌合蛋白具有抗原性而没有变应原性,分子对接表明它与 Toll 样受体 2 和 4 结合。因此,Tc24:Co1 在海洋微藻 Schizochytrium sp. 中表达,并通过酶联免疫吸附试验(ELISA)证实诱导后 48 小时表达,海藻培养物的产量为 632µg/L(冻干海藻细胞为 300µg/g)。通过口服给予表达 Tc24:Co1 的全细胞 Schizochytrium sp.(7.5µg 或 15µg 的 Tc24:Co1 剂量),在小鼠中检测到特异性血清 IgG 和肠道黏膜 IgA 反应,以及血清 Th1/Th2 细胞因子的增加。总之,表达 Tc24:Co1 的 Schizochytrium sp. 是一种有前途的口服疫苗原型,将在克氏锥虫感染的动物模型中进行评估。

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