Department of Medical Parasitology, Faculty of Medicine, Alexandria University, Alexandria, Egypt.
Department of Pathology, Faculty of Medicine, Alexandria University, Alexandria, Egypt.
PLoS Negl Trop Dis. 2021 Oct 13;15(10):e0009866. doi: 10.1371/journal.pntd.0009866. eCollection 2021 Oct.
Extracellular vesicles (EVs) are protein-loaded nano-scaled particles that are extracellularly released by eukaryotes and prokaryotes. Parasite's EVs manipulate the immune system, making them probable next-generation vaccines. Schistosomal EVs carry different proteins of promising immunizing potentials. For evaluating the immune-protective role of Schistosoma mansoni (S. mansoni) egg-derived EVs against murine schistosomiasis, EVs were isolated from cultured S. mansoni eggs by progressive sequential cooling ultra-centrifugation technique. Isolated EVs were structurally identified using transmission electron microscope and their protein was quantified by Lowry's technique. Experimental mice were subcutaneously immunized with three doses of 20 μg EVs (with or without alum adjuvant); every two weeks, then were challenged with S. mansoni cercariae two weeks after the last immunizing dose. Six weeks post infection, mice were sacrificed for vaccine candidate assessment. EVs protective efficacy was evaluated through parasitological, histopathological, and immunological parameters. Results showed significant reduction of tegumentally deranged adult worms, hepatic and intestinal egg counts reduction by 46.58%, 93.14% and 93.17% respectively, accompanied by remarkable amelioration of sizes, numbers and histopathology of hepatic granulomata mediated by high interferon gamma (IFN γ) and antibody level. Using sera from vaccinated mice, the molecular weight of EVs' protein components targeted by the antibody produced was recognized by western immunoblot. Results revealed two bands of ~ 14 KDa and ~ 21 KDa, proving that EVs are able to stimulate specific antibodies response. In conclusion, the present study highlighted the role of S. mansoni-egg derived EVs as a potential vaccine candidate against murine schistosomiasis mansoni.
细胞外囊泡 (EVs) 是由真核生物和原核生物细胞外释放的富含蛋白质的纳米级颗粒。寄生虫的 EVs 可以操纵免疫系统,使它们成为下一代疫苗的候选物。血吸虫 EVs 携带具有潜在免疫原性的不同蛋白质。为了评估曼氏血吸虫(S. mansoni)卵衍生 EVs 对小鼠血吸虫病的免疫保护作用,采用逐步连续冷却超速离心技术从培养的 S. mansoni 卵中分离 EVs。使用透射电子显微镜对分离的 EVs 进行结构鉴定,并使用 Lowry 技术定量其蛋白质。实验小鼠通过皮下免疫三次 20 μg EVs(有或没有明矾佐剂);每两周一次,最后一次免疫后两周用 S. mansoni 尾蚴进行攻击。感染后 6 周,处死小鼠进行候选疫苗评估。通过寄生虫学、组织病理学和免疫学参数评估 EVs 的保护效果。结果显示,表皮异常的成虫数量显著减少,肝和肠中的卵计数分别减少了 46.58%、93.14%和 93.17%,同时干扰素 γ (IFN γ) 和抗体水平显著升高,肝肉芽肿的大小、数量和组织病理学也得到显著改善。用免疫小鼠的血清,Western 免疫印迹显示抗体针对的 EVs 蛋白成分的分子量。结果显示两个约 14 kDa 和约 21 kDa 的条带,证明 EVs 能够刺激特异性抗体反应。综上所述,本研究强调了 S. mansoni-egg 衍生 EVs 作为一种针对曼氏血吸虫病的潜在疫苗候选物的作用。