Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Zurich, Switzerland.
Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland.
Front Cell Infect Microbiol. 2023 Mar 16;13:1162530. doi: 10.3389/fcimb.2023.1162530. eCollection 2023.
Alveolar (AE) and cystic echinococcosis (CE) are severe parasitic zoonoses caused by the larval stages of and , respectively. A panel of 7 monoclonal antibodies (mAbs) was selected against major diagnostic epitopes of both species. The binding capacity of the mAbs to spp. excretory/secretory products (ESP) was analyzed by sandwich-ELISA, where mAb Em2G11 and mAb EmG3 detected extravesicular ESP of both and These findings were subsequently confirmed by the detection of circulating ESP in a subset of serum samples from infected hosts including humans. Extracellular vesicles (EVs) were purified, and the binding to mAbs was analyzed by sandwich-ELISA. Transmission electron microscopy (TEM) was used to confirm the binding of mAb EmG3 to EVs from intravesicular fluid of spp. vesicles. The specificity of the mAbs in ELISA corresponded to the immunohistochemical staining (IHC-S) patterns performed on human AE and CE liver sections. Antigenic small particles designated as ''spems'' for and ''spegs'' for were stained by the mAb EmG3, mAb EmG3, mAb AgB, and mAb 2B2, while mAb Em2G11 reacted with spems and mAb Eg2 with spegs only. The laminated layer (LL) of both species was strongly visualized by using mAb EmG3, mAb EmG3, mAb AgB, and mAb 2B2. The LL was specifically stained by mAb Em2G11 in and by mAb Eg2 in In the germinal layer (GL), including the protoscoleces, a wide staining pattern with all structures of both species was observed with mAb EmG3, mAb EmG3, mAb AgB, mAb 2B2, and mAb Em18. In the GL and protoscoleces, the mAb Eg2 displayed a strong specific binding, while mAb Em2G11 exhibited a weak granular specific reaction. The most notable staining pattern in IHC-S was found with mAb Em18, which solely bound to the GL and protoscoleces of species and potentially to primary cells. To conclude, mAbs represent valuable tools for the visualization of major antigens in the most important species, as well as providing insights into parasite-host interactions and pathogenesis.
肺泡型（AE）和囊型包虫病（CE）是由 和 的幼虫阶段分别引起的严重寄生虫人畜共患病。针对这两个物种的主要诊断表位，选择了一组 7 种单克隆抗体（mAb）。通过夹心 ELISA 分析 mAb 对 spp. 分泌/外排产物（ESP）的结合能力，mAb Em2G11 和 mAb EmG3 检测到 和 的囊外 ESP。这些发现随后通过在包括人类在内的感染宿主的部分血清样本中检测到循环 ESP 得到证实。纯化了细胞外囊泡（EVs），并通过夹心 ELISA 分析了 mAb 的结合。透射电子显微镜（TEM）用于确认 mAb EmG3 与 spp. 囊泡腔内 EVs 的结合。ELISA 中的 mAb 特异性与在人类 AE 和 CE 肝切片上进行的免疫组织化学染色（IHC-S）模式相对应。针对 命名为“spems”的抗原小颗粒和针对 命名为“spegs”的抗原小颗粒被 mAb EmG3、mAb EmG3、mAb AgB 和 mAb 2B2 染色，而 mAb Em2G11 仅与 spems 反应，mAb Eg2 仅与 spegs 反应。使用 mAb EmG3、mAb EmG3、mAb AgB 和 mAb 2B2 强烈可视化了两种物种的层状层（LL）。mAb Em2G11 在 中特异性染色 LL，mAb Eg2 在 中特异性染色 LL。在生殖层（GL）中，包括原头蚴，用 mAb EmG3、mAb EmG3、mAb AgB、mAb 2B2 和 mAb Em18 观察到两种物种的所有结构的广泛染色模式。在 GL 和原头蚴中，mAb Eg2 显示出强烈的 特异性结合，而 mAb Em2G11 显示出微弱的颗粒状 特异性反应。在 IHC-S 中观察到的最显著染色模式是 mAb Em18，它仅与 物种的 GL 和原头蚴结合，并且可能与原代细胞结合。总之，mAb 是可视化最重要的 物种中的主要抗原的有价值的工具，并为寄生虫-宿主相互作用和发病机制提供了深入的了解。
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