Costa Marliete Carvalho, Mata Lays Murta, Ribeiro Noelly de Queiroz, Santos Anderson Philip Nonato, Oliveira Lorena Vivien Neves, Vilela Raquel Virgínia Rocha, Cardoso Valbert Nascimento, Fernandes Simone Odília Antunes, Santos Daniel Assis
Laboratório de Micologia, Departamento de Microbiologia, ICB - UFMG.
Laboratório de Radioisótopos, Departamento de Análises Clinicas e Toxicológicas, Faculdade de Farmácia-UFMG.
Med Mycol. 2018 Jun 1;56(4):479-484. doi: 10.1093/mmy/myx060.
Cryptococcus gattii is one of the etiologic agents of cryptococcosis, a systemic mycosis that occurs in healthy and immunosuppressed humans and animals worldwide. Primary pulmonary infection caused by C. gattii is usually followed by fungal dissemination to the central nervous system, resulting in high mortality rates. In this context, animal models of cryptococcosis are useful in the study of fungal pathogenesis and host response against the pathogen, and for testing novel therapeutic options. The most frequently applied method to study fungal dissemination from the lungs to other organs is by culturing tissues, which is not accurate for the detection and quantification of fungal load at early stages of the infection. To overcome this problem, the purpose of this study was to develop a new method for the quantification of Cryptococcus dissemination. One C. gattii strain was efficiently radiolabeled with technetium-99m (99mTc), without affecting viability of the cells. Further, the 99mTc-C. gattii (111 MBq) strain was used to infect mice by intratracheal and intravenous route for biodistribution studies. 99mTc-C. gattii was successfully used in detection of the yeast in the brain of mice 6 hours postinoculation, while the detection using colony forming units was possible only 24 hours postinfection. Our results provided an alternative method that could be applied in further investigations regarding the efficacy of antifungals, fungal virulence, and host-pathogen interactions.
加氏隐球菌是隐球菌病的病原体之一,隐球菌病是一种系统性真菌病,在全球范围内的健康人和免疫功能低下的人和动物中均有发生。由加氏隐球菌引起的原发性肺部感染通常随后会导致真菌扩散至中枢神经系统,从而导致高死亡率。在此背景下,隐球菌病动物模型在研究真菌发病机制和宿主对病原体的反应以及测试新型治疗方案方面很有用。研究真菌从肺部扩散到其他器官最常用的方法是培养组织,但这种方法对于感染早期真菌载量的检测和定量并不准确。为了克服这个问题,本研究的目的是开发一种新的加氏隐球菌扩散定量方法。一株加氏隐球菌菌株用99m锝(99mTc)进行了有效放射性标记,且不影响细胞活力。此外,99mTc标记的加氏隐球菌(111 MBq)菌株通过气管内和静脉途径用于感染小鼠以进行生物分布研究。接种后6小时,99mTc标记的加氏隐球菌成功用于检测小鼠脑中的酵母,而使用菌落形成单位进行检测仅在感染后24小时才可行。我们的结果提供了一种替代方法,可应用于进一步研究抗真菌药物的疗效、真菌毒力和宿主-病原体相互作用。