Mehta Sanjay R, Huang Robert, Yang Meng, Zhang Xing-Quan, Kolli Bala, Chang Kwang-Poo, Hoffman Robert M, Goto Yasuyuki, Badaro Roberto, Schooley Robert T
Department of Medicine, University of California, San Diego, San Diego, CA, USA.
Clin Vaccine Immunol. 2008 Dec;15(12):1764-70. doi: 10.1128/CVI.00270-08. Epub 2008 Oct 22.
Leishmania species are obligate intracellular protozoan parasites that cause a broad spectrum of clinical diseases in mammalian hosts. The most frequently used approach to quantify parasites in murine model systems is based on thickness measurements of the footpad or ear after experimental infection. To overcome the limitations of this method, we used a Leishmania mutant episomally transfected with enhanced green fluorescent protein, enabling in vivo real-time whole-body fluorescence imaging, to follow the progression of Leishmania infection in parasitized tissues. Fluorescence correlated with the number of Leishmania parasites in the tissue and demonstrated the real-time efficacy of a therapeutic vaccine. This approach provides several substantial advantages over currently available animal model systems for the in vivo study of immunopathogenesis, prevention, and therapy of leishmaniasis. These include improvements in sensitivity and the ability to acquire real-time data on progression and spread of the infection.
利什曼原虫属是专性细胞内原生动物寄生虫,可在哺乳动物宿主中引发多种临床疾病。在小鼠模型系统中,最常用的量化寄生虫的方法是基于实验感染后足垫或耳部的厚度测量。为克服该方法的局限性,我们使用了一种经增强型绿色荧光蛋白附加型转染的利什曼原虫突变体,从而能够进行体内实时全身荧光成像,以追踪利什曼原虫在受感染组织中的感染进程。荧光与组织中利什曼原虫的数量相关,并证明了治疗性疫苗的实时疗效。与目前用于利什曼病免疫发病机制、预防和治疗体内研究的动物模型系统相比,该方法具有几个显著优势。这些优势包括提高了灵敏度以及能够获取关于感染进程和传播的实时数据。