Red-Shifted Firefly Luciferase Optimized for Candida albicans In vivo Bioluminescence Imaging.
Red-Shifted Firefly Luciferase Optimized for Candida albicans In vivo Bioluminescence Imaging.
复制标题
DOI：
10.3389/fmicb.2017.01478
复制
发表时间：
2017
影响因子：
5.2
通讯作者：
Sanglard D
中科院分区：
文献类型：
--
作者：
Dorsaz S;Coste AT;Sanglard D
白色念珠菌是一种主要的真菌病原体，在免疫功能低下的患者中引起危及生命的疾病。目前抗C.白色念珠菌感染是有限的，因为这些感染具有40-60%的死亡率。有一个真实的需要新的治疗方法，但这样的进步需要详细的知识，C。白念珠菌及其体内发病机制。此外，任何新的抗真菌药物对C。白色念珠菌感染将需要测试它们随时间的体内效力。真菌发病机制和药物介导的解决方案的研究都可以使用非侵入性体内成像技术进行评估。在这里介绍的工作中，我们使用密码子优化的萤火虫荧光素酶报告系统检测C。小鼠中的白色念珠菌。我们调整了萤火虫荧光素酶，以提高其在红光范围（600-700 nm）的最大发射强度，以及提高其在小鼠中的热稳定性。实验动物的所有非侵入性体内成像均使用能够检测发光报告物并捕获反射率和X射线图像的多模式成像系统进行。改造后的萤火虫荧光素酶在C.发现与未修饰的荧光素酶（Mut 0）相比，白色念珠菌（Mut 2）显著增加了系统性感染中生物发光成像（BLI）的灵敏度。在口咽念珠菌病模型中使用相同的经修饰的生物发光报告系统。在这两种动物模型中，真菌负荷可能与发射光的强度相关。抗真菌治疗效果也进行了评估的基础上BLI信号强度。总之，带有红移萤火虫荧光素酶的BLI被发现是检测C.在各种小鼠感染模型中的白色念珠菌。
Candida albicans is a major fungal pathogen causing life-threatening diseases in immuno-compromised patients. The efficacy of current drugs to combat C. albicans infections is limited, as these infections have a 40–60% mortality rate. There is a real need for novel therapeutic approaches, but such advances require a detailed knowledge of C. albicans and its in vivo pathogenesis. Additionally, any novel antifungal drugs against C. albicans infections will need to be tested for their in vivo efficacy over time. Fungal pathogenesis and drug-mediated resolution studies can both be evaluated using non-invasive in vivo imaging technologies. In the work presented here, we used a codon-optimized firefly luciferase reporter system for detecting C. albicans in mice. We adapted the firefly luciferase in order to improve its maximum emission intensity in the red light range (600–700 nm) as well as to improve its thermostability in mice. All non-invasive in vivo imaging of experimental animals was performed with a multimodal imaging system able to detect luminescent reporters and capture both reflectance and X-ray images. The modified firefly luciferase expressed in C. albicans (Mut2) was found to significantly increase the sensitivity of bioluminescence imaging (BLI) in systemic infections as compared to unmodified luciferase (Mut0). The same modified bioluminescence reporter system was used in an oropharyngeal candidiasis model. In both animal models, fungal loads could be correlated to the intensity of emitted light. Antifungal treatment efficacies were also evaluated on the basis of BLI signal intensity. In conclusion, BLI with a red-shifted firefly luciferase was found to be a powerful tool for testing the fate of C. albicans in various mice infection models.