Chaimon Salisa, Phuphisut Orawan, Reamtong Onrapak, Ampawong Sumate, Fongsodsri Kamonpan, Chantree Pathanin, Thanongsaksrikul Jeeraphong, Malaithong Preeyarat, Sreesai Suthasinee, Maleewong Wanchai, Sadaow Lakkhana, Martviset Pongsakorn, Adisakwattana Poom
Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400, Thailand.
Department of Preclinical Science, Faculty of Medicine, Thammasat University, Pathumthani, 12120, Thailand.
Sci Rep. 2024 Dec 28;14(1):31229. doi: 10.1038/s41598-024-82599-x.
The cytokine homologs, particularly transforming growth factor (TGF)-β, is a crucial immunomodulatory molecule and involved in growth and developmental processes in several helminths. In this study, the basic properties and functions of T. spiralis TGF-β homolog 2 (TsTGH2) were characterized using bioinformatics and molecular biology approaches. Bioinformatics analyses indicated that TsTGH2 belongs to the TGF-β subfamily. Recombinant TsTGH2 (rTsTGH2) expressed in Escherichia coli was used to produce a polyclonal antibody (pAb) in mice. Western blot and immunolocalization using pAb detected native TsTGH2 in crude worm antigens from muscle larvae and adults, showing it was mainly localized in the body wall muscles and the epithelia of the ovary and uterus. To assess the interplay between TsTGH2 and the human TGF-β signaling pathway, rTsTGH2 produced in a HEK293T cell was incubated with the SBE luciferase-HEK293 cell. The result indicated a significant increase in luciferase activity after treatment with rTsTGH2 compared to untreated control (p < 0.05). In conclusion, these findings are the first to characterize the basic properties and functions of TGF-β homologs in T. spiralis, demonstrating their interaction with the human TGF-β receptor. Further investigation is required to identify and optimize an appropriate expression system or conditions for TsTGH2. Additionally, studies are needed to clarify the specific role of native TsTGH2 in parasite development and host immunomodulation.
细胞因子同源物,特别是转化生长因子(TGF)-β,是一种关键的免疫调节分子,参与多种蠕虫的生长和发育过程。在本研究中,利用生物信息学和分子生物学方法对旋毛虫TGF-β同源物2(TsTGH2)的基本特性和功能进行了表征。生物信息学分析表明,TsTGH2属于TGF-β亚家族。在大肠杆菌中表达的重组TsTGH2(rTsTGH2)用于在小鼠中制备多克隆抗体(pAb)。使用pAb进行的蛋白质免疫印迹和免疫定位检测到来自肌幼虫和成虫的粗虫抗原中的天然TsTGH2,表明其主要定位于体壁肌肉以及卵巢和子宫的上皮细胞。为了评估TsTGH2与人类TGF-β信号通路之间的相互作用,将在HEK293T细胞中产生的rTsTGH2与SBE荧光素酶-HEK293细胞一起孵育。结果表明,与未处理的对照相比,用rTsTGH2处理后荧光素酶活性显著增加(p < 0.05)。总之,这些发现首次表征了旋毛虫中TGF-β同源物的基本特性和功能,证明了它们与人类TGF-β受体的相互作用。需要进一步研究以鉴定和优化TsTGH2的合适表达系统或条件。此外,还需要开展研究以阐明天然TsTGH2在寄生虫发育和宿主免疫调节中的具体作用。