Abstract：
BACKGROUND: Mucormycosis is an aggressive, invasive fungal infection caused by moulds in the order Mucorales. Early diagnosis is key to improving patient prognosis, yet relies on insensitive culture or non-specific histopathology. A pan-Mucorales specific monoclonal antibody (mAb), TG11, was recently developed. Here, we investigate the spatio-temporal localisation of the antigen and specificity of the mAb for immunohistochemistry.
METHODS: We use immunofluorescence (IF) microscopy to assess antigen localisation in eleven Mucorales species of clinical importance and live imaging of Rhizopus arrhizus germination. Immunogold transmission electron microscopy (immunoTEM) reveals the sub-cellular location of mAb TG11 binding. Finally, we perform immunohistochemistry of R. arrhizus in an ex vivo murine lung infection model alongside lung infection by Aspergillus fumigatus.
RESULTS: IF revealed TG11 antigen production at the emerging hyphal tip and along the length of growing hyphae in all Mucorales except Sakasenea. Timelapse imaging revealed early antigen exposure during spore germination and along the growing hypha. ImmunoTEM confirmed mAb TG11 binding to the hyphal cell wall only. The TG11 mAb specifically stained Mucorales but not Aspergillus hyphae in infected murine lung tissue.
CONCLUSIONS: TG11 detects early hyphal growth and has valuable potential for diagnosing mucormycosis by enhancing discriminatory detection of Mucorales in tissue.
METHODS: We use immunofluorescence (IF) microscopy to assess antigen localisation in eleven Mucorales species of clinical importance and live imaging of Rhizopus arrhizus germination. Immunogold transmission electron microscopy (immunoTEM) reveals the sub-cellular location of mAb TG11 binding. Finally, we perform immunohistochemistry of R. arrhizus in an ex vivo murine lung infection model alongside lung infection by Aspergillus fumigatus.
RESULTS: IF revealed TG11 antigen production at the emerging hyphal tip and along the length of growing hyphae in all Mucorales except Sakasenea. Timelapse imaging revealed early antigen exposure during spore germination and along the growing hypha. ImmunoTEM confirmed mAb TG11 binding to the hyphal cell wall only. The TG11 mAb specifically stained Mucorales but not Aspergillus hyphae in infected murine lung tissue.
CONCLUSIONS: TG11 detects early hyphal growth and has valuable potential for diagnosing mucormycosis by enhancing discriminatory detection of Mucorales in tissue.
摘要：
背景：毛霉菌病是一种侵袭性的，由霉菌引起的侵袭性真菌感染。早期诊断是改善患者预后的关键,然而依赖于不敏感的培养或非特异性组织病理学。一种泛Mucorales特异性单克隆抗体（mAb），TG11是最近开发的。这里,我们研究了抗原的时空定位和mAb的免疫组织化学特异性。
方法：我们使用免疫荧光（IF）显微镜来评估11种具有临床重要性的毛霉菌中的抗原定位，并对根霉萌发进行实时成像。免疫金透射电子显微镜(免疫TEM)揭示了mAbTG11结合的亚细胞位置。最后,我们在离体小鼠肺部感染模型以及烟曲霉肺部感染中进行了rhizus的免疫组织化学。
结果：IF显示，除Sakasenea外，所有Mucorales中，TG11抗原在新兴菌丝尖端和生长菌丝的长度上产生。时间流逝成像显示，在孢子萌发和沿着菌丝生长的过程中，早期抗原暴露。免疫TEM证实mAbTG11仅与菌丝细胞壁结合。TG11mAb在受感染的鼠肺组织中特异性地染色Mucorales而不是曲霉菌丝。
结论：TG11检测早期菌丝生长，通过增强组织中毛菌病的鉴别检测，对诊断毛霉菌病具有有价值的潜力。
方法：我们使用免疫荧光（IF）显微镜来评估11种具有临床重要性的毛霉菌中的抗原定位，并对根霉萌发进行实时成像。免疫金透射电子显微镜(免疫TEM)揭示了mAbTG11结合的亚细胞位置。最后,我们在离体小鼠肺部感染模型以及烟曲霉肺部感染中进行了rhizus的免疫组织化学。
结果：IF显示，除Sakasenea外，所有Mucorales中，TG11抗原在新兴菌丝尖端和生长菌丝的长度上产生。时间流逝成像显示，在孢子萌发和沿着菌丝生长的过程中，早期抗原暴露。免疫TEM证实mAbTG11仅与菌丝细胞壁结合。TG11mAb在受感染的鼠肺组织中特异性地染色Mucorales而不是曲霉菌丝。
结论：TG11检测早期菌丝生长，通过增强组织中毛菌病的鉴别检测，对诊断毛霉菌病具有有价值的潜力。
