State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, China.
Front Immunol. 2019 Jul 23;10:1707. doi: 10.3389/fimmu.2019.01707. eCollection 2019.
Cathepsin B, a lysosomal cysteine protease, is thought to be involved in the pathogenesis of infection, but its exact role remains unclear. In the present study, a recombinant cathepsin B (rFgCatB) protein was expressed in the methylotrophic yeast . Western blot analysis confirmed the reactivity of the purified rFgCatB protein to serum from -infected goats. The effects of serial concentrations (10, 20, 40, 80, and 160 μg/ml) of rFgCatB on various functions of goat peripheral blood mononuclear cells (PBMCs) were examined. We demonstrated that rFgCatB protein can specifically bind to the surface of PBMCs. In addition, rFgCatB increased the expression of cytokines (IL-2, IL-4, IL-10, IL-17, TGF-β, and IFN-γ), and increased nitric oxide production and cell apoptosis, but reduced cell viability. These data show that rFgCatB can influence cellular and immunological functions of goat PBMCs. Further characterization of the posttranslational modification and assessment of rFgCatB in immunogenicity studies is warranted.
组织蛋白酶 B（Cathepsin B）是一种溶酶体半胱氨酸蛋白酶，被认为与 感染的发病机制有关，但确切作用仍不清楚。本研究在甲醇营养型酵母中表达了重组 组织蛋白酶 B（rFgCatB）蛋白。Western blot 分析证实了纯化的 rFgCatB 蛋白对 感染山羊血清的反应性。检测了 rFgCatB 的系列浓度（10、20、40、80 和 160 μg/ml）对山羊外周血单个核细胞（PBMCs）各种功能的影响。结果表明，rFgCatB 蛋白可以特异性结合 PBMCs 表面。此外，rFgCatB 增加了细胞因子（IL-2、IL-4、IL-10、IL-17、TGF-β 和 IFN-γ）的表达，增加了一氧化氮的产生和细胞凋亡，但降低了细胞活力。这些数据表明，rFgCatB 可以影响山羊 PBMCs 的细胞和免疫功能。需要进一步研究 rFgCatB 的翻译后修饰，并评估其在免疫原性研究中的作用。
