ICMR-National Institute of Traditional Medicine, Belagavi 590010, Karnataka, India.
ICMR-National Institute of Traditional Medicine, Belagavi 590010, Karnataka, India.
Int J Biol Macromol. 2020 May 1;150:206-217. doi: 10.1016/j.ijbiomac.2020.02.024. Epub 2020 Feb 5.
Galectins regulate growth and differentiation of immune cells and inflammation through their carbohydrate-binding function in humans, while also play a role in parasite survival. This study focused on the galectin of lymphatic filarial parasite Wuchereria bancrofti (Wb-Gal). The multiple sequence alignment with other galectins showed that the Wb-Gal belonged to galactoside binding lectin family, particularly tandem repeat type galectin-9. A homology model of Wb-Gal was developed in the I-TASser server using high similarity 3D structures with a quality score of 89.5. Molecular docking and dynamics studies revealed that the CCRD and NCRD of Wb-Gal bind with galactose and lactose. Further, Wb-Gal was cloned into the pET28 vector, expressed in E. coli Rosetta strain and purified by affinity chromatography. In the hemagglutination assays, the rWb-Gal bound to lactose, galactose, and glucose. Indirect Enzyme-Linked Immunosorbent Assay (ELISA) using different clinical filarial sera showed that the IgG and IgM response was against Wb-Gal x very high in all filarial clinical groups, whereas the IgA and IgG2 response was minimum to negligible. There was an enhanced response of IgG1 and IgG4 antibodies in Microfilaremics (MF) cases compared to Chronic Pathology (CP) and Endemic Normal (EN) individuals. Interestingly, the IgE response was comparatively higher in EN than MF and CP. These studies show that Wb-Gal is a member of the lectin family of proteins binding to different carbohydrates and may have an important role in the pathophysiology of filarial infection which needs to be investigated in greater detail.
半乳糖凝集素通过其在人类中的碳水化合物结合功能调节免疫细胞的生长和分化以及炎症,同时在寄生虫存活中发挥作用。本研究专注于淋巴丝虫寄生虫班氏吴策线虫的半乳糖凝集素(Wb-Gal)。与其他半乳糖凝集素的多重序列比对表明,Wb-Gal 属于半乳糖结合凝集素家族,特别是串联重复型半乳糖凝集素-9。使用与高质量分数 89.5 的高相似度 3D 结构,在 I-TASser 服务器中开发了 Wb-Gal 的同源模型。分子对接和动力学研究表明,Wb-Gal 的 CCRD 和 NCRD 与半乳糖和乳糖结合。此外,将 Wb-Gal 克隆到 pET28 载体中,在大肠杆菌 Rosetta 菌株中表达并通过亲和层析纯化。在血凝试验中,rWb-Gal 与乳糖、半乳糖和葡萄糖结合。使用不同的临床丝虫血清进行间接酶联免疫吸附试验(ELISA)表明,针对 Wb-Gal 的 IgG 和 IgM 反应在所有丝虫临床组中均非常高,而 IgA 和 IgG2 反应则最小或可忽略不计。微丝蚴(MF)病例中的 IgG1 和 IgG4 抗体的反应增强,与慢性病理(CP)和流行正常(EN)个体相比。有趣的是,与 MF 和 CP 相比,EN 中的 IgE 反应相对较高。这些研究表明,Wb-Gal 是一种结合不同碳水化合物的凝集素家族蛋白的成员,可能在丝虫感染的病理生理学中具有重要作用,需要更详细地研究。