Duncan Joshua, Cutress David, Morphew Russell M, Brophy Peter M
Institute of Biological, Environmental and Rural Sciences (IBERS), Aberystwyth University, Aberystwyth SY23 3DA, UK.
Institute of Biological, Environmental and Rural Sciences (IBERS), Aberystwyth University, Aberystwyth SY23 3DA, UK.
Mol Biochem Parasitol. 2018 Jun;222:45-50. doi: 10.1016/j.molbiopara.2018.04.006. Epub 2018 Apr 27.
Fascioliasis is a parasitic disease of grazing livestock and a threat to global food security by significantly reducing the production value of sheep, goats and cattle. Moreover, the zoonotic parasite is also a re-emerging food borne threat to human populations. Driven by climate change, the prevalence of fascioliasis is set to increase. Efforts to control the main causative agent, Fasciola hepatica, are hampered by short lived chemotherapy approaches that are becoming increasingly obsolete due to therapeutic failure and resistance. A protective vaccine is urgently needed. A recombinant form of Sigma class glutathione transferase (Hematopoietic Prostaglandin D synthase) from F. hepatica (FhGSTS1) with confirmed prostaglandin synthase activity shows immune-modulation activity via suppression of Th17 responses in host dendritic cells. In vaccine trials recombinant FhGSTS1 reduces liver pathology but not worm burden. Native FhGSTS1 is yet to be tested for immune-modulation activities or for vaccine potential, primarily due to the technical difficulty in purifying FhGST-S1 away from the other more abundant GST members in F. hepatica cytosol. This paper reports a pipeline for the purification of native FhGSTS1 using a two-step process consisting of glutathione-agarose affinity and cationic exchange chromatography. The methodology allows for the isolation of purified and active FhGSTS1 or Sigma GSTs from other sources for analytical biochemical and immunological studies.
肝片吸虫病是一种放牧家畜的寄生虫病，通过显著降低绵羊、山羊和牛的产值，对全球粮食安全构成威胁。此外，这种人畜共患寄生虫也是一种新出现的食源性人类健康威胁。在气候变化的推动下，肝片吸虫病的流行率将上升。由于化疗方法寿命短，且因治疗失败和耐药性而日益过时，控制主要病原体肝片吸虫（Fasciola hepatica）的努力受到阻碍。迫切需要一种保护性疫苗。来自肝片吸虫的重组形式的西格玛类谷胱甘肽转移酶（造血前列腺素D合成酶，FhGSTS1）具有已确认的前列腺素合成酶活性，通过抑制宿主树突状细胞中的Th17反应显示出免疫调节活性。在疫苗试验中，重组FhGSTS1可减轻肝脏病理，但不能减轻虫负荷。天然FhGSTS1尚未进行免疫调节活性或疫苗潜力测试，主要是由于从肝片吸虫细胞质中其他更丰富的谷胱甘肽转移酶成员中纯化FhGST-S1存在技术困难。本文报道了一种使用两步法纯化天然FhGSTS1的流程，该方法包括谷胱甘肽-琼脂糖亲和层析和阳离子交换层析。该方法可从其他来源分离纯化的活性FhGSTS1或西格玛谷胱甘肽转移酶，用于分析生物化学和免疫学研究。
