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. 2020 Dec;59(1):e127.
doi: 10.1002/cpmc.127.

A Mouse Model of Sublethal Leptospirosis: Protocols for Infection with Leptospira Through Natural Transmission Routes, for Monitoring Clinical and Molecular Scores of Disease, and for Evaluation of the Host Immune Response

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A Mouse Model of Sublethal Leptospirosis: Protocols for Infection with Leptospira Through Natural Transmission Routes, for Monitoring Clinical and Molecular Scores of Disease, and for Evaluation of the Host Immune Response

Nisha Nair et al. Curr Protoc Microbiol. 2020 Dec.

Abstract

Leptospirosis is a zoonotic disease caused by pathogenic Leptospira species that are maintained in sylvatic and domestic environments by transmission among rodents and other carriers. Humans become infected after contact of breached skin or mucosa with contaminated water or soil. Understanding persistent or sublethal infection in a host is critical for controlling human risk of exposure to pathogenic Leptospira. Animal models that recapitulate disease progression after infection via natural transmission routes are more appropriate for validation of vaccines and therapeutics. Furthermore, the ability to measure shedding of live Leptospira in urine of reservoir and carrier hosts can be used to develop new diagnostic assays and sensors to evaluate human risk of exposure. We developed inbred mouse models of Leptospirosis, that bypass survival as a criterion, in which we can analyze both pathogen and host factors affecting sublethal infection (<1 month), including shedding of Leptospira in urine. Mice are infected with pathogenic Leptospira using a physiologic route, and the clinical, histological, and molecular scores of disease are measured. Furthermore, the host immune response to Leptospira is evaluated. This mouse model also provides a tool in which to test fundamental hypotheses related to host-pathogen interactions and the immune mechanisms engaged in protective and pathogenic immune responses. © 2020 Wiley Periodicals LLC Basic Protocol 1: Culture and maintenance of virulent Leptospira Basic Protocol 2: Infection of mice through a physiologic route and collection of clinical scores and biological samples Basic Protocol 3: Analysis of pathogenesis after Leptospira infection.

Keywords: Leptospira; mouse model; natural transmission routes; physiologic route of infection; sublethal leptospirosis.

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Figures

Figure 1.
Figure 1.
Workflow for infection of mice with pathogenic Leptospira and subsequent collection of biological samples.
Figure 2.
Figure 2.
Histopathology of the kidney. (A) PAS-D staining showing shrinkage of glomeruli, infiltration of immune cells and loss of tubular structure in the infected group (20 X); (B) diameter of glomeruli is measured under 20 X using a measurement function (ex. CaptaVision Software). Legend: white arrow, tubules; black arrow, infiltration of immune cells; black triangle, glomerulus (Sullivan, Nair et al. 2017).
Figure 3.
Figure 3.
Histopathology of the kidney. (A) Masson trichrome staining of kidney sections from infected mice pre-treated with PBS and with L. plantarum. B. Digital quantification of fibrosis was determined as the % area (pixels) where blue staining exceeds a threshold. The slides were processed and stained in parallel and images where taken using the same illumination (Potula, Richer et al. 2017).
Figure 4.
Figure 4.
Immunohistochemistry of the kidney. Immunostaining of kidney sections from groups of treated mice in the presence or absence of Leptospira infection using various leucocyte markers (CD45+, NIMP-R14+, F4/80+ and CD3+) (Potula, Richer et al. 2017).
Figure 5.
Figure 5.
Susceptibility to lethal leptospirosis (LD50) in C3H-HeJ mice inoculated with PBS (Ctrl), 5×108 and 1×109 Leptospira interrogans ser. Copenhageni FioCruz. N=4 mice per group.

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