Melo Guilherme D, Goyard Sophie, Lecoeur Hervé, Rouault Eline, Pescher Pascale, Fiette Laurence, Boissonnas Alexandre, Minoprio Paola, Lang Thierry
Institut Pasteur, Laboratoire des Processus Infectieux à Trypanosomatidés, Département Infection et Epidémiologie, 25-28 rue du Dr Roux, Paris, France.
Institut Pasteur, Unité de Parasitologie Moléculaire et Signalisation, Département de Parasites et Insectes Vecteurs, 25-28 rue du Dr Roux, Paris, France.
PLoS Negl Trop Dis. 2017 Sep 25;11(9):e0005924. doi: 10.1371/journal.pntd.0005924. eCollection 2017 Sep.
Visceral leishmaniasis is an insidious neglected disease with worldwide distribution. It is caused by parasites from the Leishmania donovani complex, which are able to be transmitted by different species of phlebotomine sand flies and to infect numerous mammal hosts. Despite the high number of people infected or at risk, and the remarkable quantity of studies focusing on this disease, a proper experimental model to efficiently decipher the infectious process of visceral leishmaniasis taking into account the nuances of parasite’s virulence and the duration of the infection is still lacking. Therefore, using golden Syrian hamsters and BALB/c mice, state-of-the-art genetic manipulation applied on a fully virulent L. donovani strain and in vivo imaging approaches, we describe herein three benefits for experimental visceral leishmaniasis: (i) the development of a double transfected bioluminescent (firefly luciferase) and fluorescent (E2-crimson) virulent strain of L. donovani (Ld1S_luci_E2-crimson), favoring a wide range of both in vivo and in vitro investigations, (ii) the establishment of a non-invasive mouse model to evaluate the infectious process during visceral leishmaniasis and the parasite’s virulence in real time, allowing longitudinal studies with the same animals, and (iii) the elaboration of a suitable method to reinstate (and verify anew) the virulence in a population of attenuated parasites, by recovering persistent parasites from chronic infected mice. Consequently, these results open up new perspectives on the study of visceral leishmaniasis, especially in the fields of therapeutics and vaccinology, since the model described herein renders now possible long-lasting follow up studies, with easy and accurate day-by-day verifications of the infection status along with a reduced number of laboratory animals.
ClinicalTrials.gov 2013-0047.
内脏利什曼病是一种分布于全球的隐匿性被忽视疾病。它由杜氏利什曼原虫复合体的寄生虫引起，这些寄生虫可通过不同种类的白蛉传播，并感染众多哺乳动物宿主。尽管感染人数众多或面临感染风险，且针对该疾病的研究数量可观，但仍缺乏一个合适的实验模型来有效解读内脏利什曼病的感染过程，同时考虑到寄生虫毒力的细微差别和感染持续时间。因此，我们使用叙利亚金黄地鼠和BALB/c小鼠，对一种完全有毒力的杜氏利什曼原虫菌株应用最先进的基因操作技术，并采用体内成像方法，在此描述了实验性内脏利什曼病的三个优势：（i）构建了一种双转染的生物发光（萤火虫荧光素酶）和荧光（E2-深红色）的杜氏利什曼原虫有毒力菌株（Ld1S_luci_E2-深红色），有利于进行广泛的体内和体外研究；（ii）建立了一种非侵入性小鼠模型，用于实时评估内脏利什曼病的感染过程和寄生虫毒力，允许对同一动物进行纵向研究；（iii）制定了一种合适的方法，通过从慢性感染小鼠中回收持续性寄生虫，恢复（并重新验证）减毒寄生虫群体的毒力。因此，这些结果为内脏利什曼病的研究开辟了新的视角，特别是在治疗学和疫苗学领域，因为本文所述模型现在使得长期随访研究成为可能，能够轻松、准确地逐日核实感染状态，同时减少实验动物数量。
ClinicalTrials.gov 2013 - 0047。
