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Abstract：
Free-living amoeba of the genus Acanthamoeba are ubiquitous protozoa involved in opportunistic and non-opportunistic infection in humans, such as granulomatous amoebic encephalitis and amoebic keratitis. Both infections have challenging characteristics such as the formation of the resistant cysts in infected tissues, hampering the treatment and most usual diagnosis depending on time-consuming and/or low sensitivity techniques. The use of monoclonal antibodies presents itself as an opportunity for the development of more effective alternative diagnostic methods, as well as an important and useful tool in the search for new therapeutic targets. This study investigated the possibility of using a previously produced monoclonal antibody (mAb3), as a diagnostic tool for the detection of Acanthamoeba trophozoites by direct and indirect flow cytometry and immunofluorescence. Immunoprecipitation assay and mass spectrometry allowed the isolation of the antibody\'s target and suggested it is a transporter part of the CPA (cation: proton antiporter) superfamily. In vitro tests indicate an important role of this target in Acanthamoeba\'s encystment physiology. Our results support the importance of studying the role of CPA2 transporters in the context of acanthamoebiasis, as this may be a way to identify new therapeutic candidates.
摘要：
棘阿米巴属的自由生活变形虫是参与人类机会性和非机会性感染的无处不在的原生动物，如肉芽肿性阿米巴性脑炎和阿米巴性角膜炎。两种感染都具有挑战性的特征，例如在感染组织中形成抗性囊肿，根据耗时和/或低灵敏度的技术阻碍治疗和最常见的诊断。单克隆抗体的使用为开发更有效的替代诊断方法提供了机会。以及寻找新的治疗靶点的重要和有用的工具。这项研究调查了使用先前生产的单克隆抗体（mAb3）的可能性，作为通过直接和间接流式细胞术和免疫荧光检测棘阿米巴滋养体的诊断工具。免疫沉淀测定和质谱允许分离抗体的靶标，并表明它是CPA（阳离子：质子反转运蛋白）超家族的转运蛋白部分。体外试验表明，该靶标在棘阿米巴的体内生理中具有重要作用。我们的结果支持研究CPA2转运蛋白在棘阿米巴病中的作用的重要性，因为这可能是一种确定新的候选治疗方法的方法。
