Li Muxiao, Ao Yangsiqi, Guo Jiaying, Nie Zheng, Liu Qin, Yu Long, Luo Xiaoying, Zhan Xueyan, Zhao Yangnan, Wang Sen, An Xiaomeng, He Lan, Zhao Junlong
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.
Key Laboratory of Preventive Veterinary Medicine in Hubei Province, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, China.
Front Microbiol. 2020 Jan 15;10:3046. doi: 10.3389/fmicb.2019.03046. eCollection 2019.
, a tick-borne intraerythrocytic zoonotic protozoan, causes most of human babesiosis in the world, and patients usually experience intermittent fever, fatigue, and chills, followed by a combination of additional symptoms and even death in severe cases. Unfortunately, there is no curable drug or effective vaccine available, and the mechanism of related virulence factors in invasion to host cells during the merozoite stage is unclear. Here, we evaluated a secreted protein annotated as surface antigen 1 (BmSA1) and identified from culture supernatant by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). BmSA1 fragment was expressed in to prepare polyclonal antiserum. Western blot analysis revealed the existence of BmSA1 in the lysate of the parasites and the hemolysate of infected red blood cells (iRBCs). Laser confocal microscopy confirmed BmSA1 as a secreted protein with diffuse distribution around the parasites in red blood cells (RBCs). The adhesion capacity of BmSA1 against the host RBCs was tested by RBC binding assays using the recombinant BmSA1 protein (rBmSA1), which was shown to specifically bind to host RBCs. Further antiserum-neutralization test demonstrated that the growth of parasites could be significantly inhibited by the anti-BmSA1 antiserum. These results indicate that BmSA1 is a crucial factor for invasion into host RBCs with an important role in host-parasite interactions during the merozoite stage and has the potential use as a vaccine candidate due to its high secretion amount.
巴贝斯虫是一种由蜱传播的胞内寄生性人兽共患原生动物,引起世界上大多数人类巴贝斯虫病,患者通常会出现间歇性发热、疲劳和寒战,随后会出现一系列其他症状,严重时甚至会死亡。不幸的是,目前尚无治愈药物或有效疫苗,并且裂殖子阶段相关毒力因子侵入宿主细胞的机制尚不清楚。在此,我们评估了一种注释为表面抗原1(BmSA1)的分泌蛋白,该蛋白通过液相色谱-串联质谱(LC-MS/MS)从培养上清液中鉴定出来。在大肠杆菌中表达BmSA1片段以制备多克隆抗血清。蛋白质免疫印迹分析显示寄生虫裂解物和感染红细胞(iRBCs)的溶血产物中存在BmSA1。激光共聚焦显微镜证实BmSA1是一种分泌蛋白,在红细胞(RBCs)中寄生虫周围呈弥漫性分布。使用重组BmSA1蛋白(rBmSA1)通过红细胞结合试验测试了BmSA1对宿主RBCs的粘附能力,结果表明它能特异性结合宿主RBCs。进一步的抗血清中和试验表明,抗BmSA1抗血清可显著抑制寄生虫的生长。这些结果表明,BmSA1是巴贝斯虫侵入宿主RBCs的关键因素,在裂殖子阶段的宿主-寄生虫相互作用中起重要作用,并且由于其高分泌量而具有作为疫苗候选物的潜力。