In vivo imaging of mice infected with bioluminescent Trypanosoma cruzi unveils novel sites of infection

Particle and Fibre Toxicology

Wanderley de Souza⁴  Aline Salgado Ribeiro¹  Marcelo Meuser-Batista³  Andréa Henriques-Pons¹  Cristina Henriques² 

[1] Laboratório de Inovações em Terapias, Ensino e Bioprodutos, Instituto Oswaldo Cruz-Fundação Oswaldo Cruz, Janeiro-RJ, Brazil;Fundação Oswaldo Cruz, Cruz- FIOCRUZ, Mato Grosso do Sul, Campo Grande-MS, Brazil;Departamento de Anatomia Patológica e Citopatologia, Instituto Fernandes Figueira-FIOCRUZ, Janeiro-RJ, Brazil;Instituto Nacional de Metrologia, Qualidade e Tecnologia-Inmetro, Janeiro-RJ, Brazil

关键词: Chagas disease;    Mice;    Heart;    Rectum;    Testicles;    Bioluminescence;    Luciferase;    Trypanosoma cruzi;   

Others  :  811877
DOI  :  10.1186/1756-3305-7-89

 received in 2013-09-20, accepted in 2014-02-21,  发布年份 2014

【 摘 要 】

Background

The development of techniques that allow the imaging of animals infected with parasites expressing luciferase opens up new possibilities for following the fate of parasites in infected mammals.

Methods

D-luciferin potassium salt stock solution was prepared in phosphate-buffered saline (PBS) at 15 mg/ml. To produce bioluminescence, infected and control mice received an intraperitoneal injection of luciferin stock solution (150 mg/kg). All mice were immediately anesthetized with 2% isofluorane, and after 10 minutes were imaged. Ex vivo evaluation of infected tissues and organs was evaluated in a 24-well plate in 150 μg/ml D-luciferin diluted in PBS. Images were captured using the IVIS Lumina image system (Xenogen). Dissected organs were also evaluated by microscopy of hematoxylin-eosin stained sections.

Results

Here we describe the results obtained using a genetically modified Dm28c strain of T. cruzi expressing the firefly luciferase to keep track of infection by bioluminescence imaging. Progression of infection was observed in vivo in BALB/c mice at various intervals after infection with transgenic Dm28c-luc. The bioluminescent signal was immediately observed at the site of T. cruzi inoculation, and one day post infection (dpi) it was disseminated in the peritoneal cavity. A similar pattern in the cavity was observed on 7 dpi, but the bioluminescence was more intense in the terminal region of the large intestine, rectum, and gonads. On 14 and 21 dpi, bioluminescent parasites were also observed in the heart, snout, paws, hind limbs, and forelimbs. From 28 dpi to 180 dpi in chronically infected mice, bioluminescence declined in regions of the body but was concentrated in the gonad region. Ex vivo evaluation of dissected organs and tissues by bioluminescent imaging confirmed the in vivo bioluminescent foci. Histopathological analysis of dissected organs demonstrated parasite nests at the rectum and snout, in muscle fibers of mice infected with Dm28c-WT and with Dm28c-luc, corroborating the bioluminescent imaging.

Conclusion

Bioluminescence imaging is accurate for tracking parasites in vivo, and this methodology is important to gain a better understanding of the infection, tissue inflammation, and parasite biology regarding host cell interaction, proliferation, and parasite clearance to subpatent levels.

【 授权许可】

   
2014 Henriques et al.; licensee BioMed Central Ltd.

【 图 表 】

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