Journal article
In vivo Imaging of Transgenic Leishmania Parasites in a Live Host
Journal of Visualized Experiments, Vol.41, e1980
2010
DOI: 10.3791/1980
PMCID: PMC3156083
PMID: 20689512
Abstract
Distinct species of Leishmania, a protozoan parasite of the family Trypanosomatidae, typically cause different human disease manifestations. The most common forms of disease are visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL). Mouse models of leishmaniasis are widely used, but quantification of parasite burdens during murine disease requires mice to be euthanized at various times after infection. Parasite loads are then measured either by microscopy, limiting dilution assay, or qPCR amplification of parasite DNA. The in vivo imaging system (IVIS) has an integrated software package that allows the detection of a bioluminescent signal associated with cells in living organisms. Both to minimize animal usage and to follow infection longitudinally in individuals, in vivo models for imaging Leishmania spp. causing VL or CL were established. Parasites were engineered to express luciferase, and these were introduced into mice either intradermally or intravenously. Quantitative measurements of the luciferase driving bioluminescence of the transgenic Leishmania parasites within the mouse were made using IVIS. Individual mice can be imaged multiple times during longitudinal studies, allowing us to assess the inter-animal variation in the initial experimental parasite inocula, and to assess the multiplication of parasites in mouse tissues. Parasites are detected with high sensitivity in cutaneous locations. Although it is very likely that the signal (photons/second/parasite) is lower in deeper visceral organs than the skin, but quantitative comparisons of signals in superficial versus deep sites have not been done. It is possible that parasite numbers between body sites cannot be directly compared, although parasite loads in the same tissues can be compared between mice. Examples of one visceralizing species (L. infantum chagasi) and one species causing cutaneous leishmaniasis (L. mexicana) are shown. The IVIS procedure can be used for monitoring and analyzing small animal models of a wide variety of Leishmania species causing the different forms of human leishmaniasis.
Details
- Title: Subtitle
- In vivo Imaging of Transgenic Leishmania Parasites in a Live Host
- Creators
- Colin J Thalhofer - Interdisciplinary Immunology Program, University of Iowa, and the VA Medical CenterJoel W Graff - Department of Biochemistry, University of Iowa, and the VA Medical CenterLaurie Love-Homan - Department of Internal Medicine, University of IowaSuzanne M Hickerson - Department of Molecular Microbiology, Washington University School of MedicineNoah Craft - Division of Dermatology, Harbor-UCLA Medical Center, Hanley-Hardison Research CenterStephen M Beverley - Department of Molecular Microbiology, Washington University School of MedicineMary E Wilson - Departments of Internal Medicine, Microbiology and Epidemiology, University of Iowa
- Resource Type
- Journal article
- Publication Details
- Journal of Visualized Experiments, Vol.41, e1980
- Publisher
- MyJove Corporation
- DOI
- 10.3791/1980
- PMID
- 20689512
- PMCID
- PMC3156083
- ISSN
- 1940-087X
- eISSN
- 1940-087X
- Language
- English
- Date published
- 2010
- Academic Unit
- Microbiology and Immunology; International Programs; Epidemiology; Internal Medicine
- Record Identifier
- 9984001145402771
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