Department of Infection Biology, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom.
Curr Pharm Des. 2021;27(14):1733-1740. doi: 10.2174/1381612826666201124113214.
Chagas disease results from infection with the trypanosomatid parasite Trypanosoma cruzi. Progress in developing new drugs has been hampered by the long term and complex nature of the condition and by our limited understanding of parasite biology. Technical difficulties in assessing the parasite burden during the chronic stage of infection have also proven to be a particular challenge. In this context, the development of noninvasive, highly sensitive bioluminescence imaging procedures based on parasites that express a red-shifted luciferase has greatly enhanced our ability to monitor infections in experimental models. Applications of this methodology have led to new insights into tissue tropism and infection dynamics and have been a major driver in drug development. The system has been further modified by the generation of parasite reporter lines that express bioluminescent:fluorescent fusion proteins, an advancement that has allowed chronic infections in mice to be examined at a cellular level. By exploiting bioluminescence, to identify the rare sites of tissue infection, and fluorescence to detect T. cruzi at the level of individual host cells in histological sections, it has been possible to investigate the replication and differentiation status of parasites in vivo and to examine the cellular environment of infection foci. In combination, these data provide a framework for the detailed dissection of disease pathogenesis and drug activity.
克氏锥虫感染导致恰加斯病。由于该病的长期性和复杂性以及我们对寄生虫生物学的有限了解，新药物的研发进展一直受到阻碍。在慢性感染阶段评估寄生虫负担的技术困难也被证明是一个特别的挑战。在这种情况下，基于表达红移荧光素酶的寄生虫的非侵入性、高灵敏度生物发光成像程序的开发极大地增强了我们在实验模型中监测感染的能力。该方法的应用为组织嗜性和感染动力学提供了新的见解，并成为药物开发的主要驱动力。该系统通过生成表达生物发光：荧光融合蛋白的寄生虫报告系进一步得到了改进，这一进展使得可以在细胞水平上检查小鼠的慢性感染。通过利用生物发光来识别组织感染的罕见部位，以及利用荧光在组织切片中检测单个宿主细胞中的克氏锥虫，可以在体内研究寄生虫的复制和分化状态，并检查感染灶的细胞环境。这些数据结合在一起，为详细剖析疾病发病机制和药物活性提供了一个框架。
