In Vivo Bioluminescence Imaging for Longitudinal Monitoring of Inflammation in Animal Models of Uveitis

Abstract

PurposeWe develop a quantitative bioluminescence assay for in vivo longitudinal monitoring of inflammation in animal models of uveitis.MethodsThree models of experimental uveitis were induced in C57BL/6 albino mice: primed mycobacterial uveitis (PMU), endotoxin-induced uveitis (EIU), and experimental autoimmune uveitis (EAU). Intraperitoneal injection of luminol sodium salt, which emits light when oxidized, provided the bioluminescence substrate. Bioluminescence images were captured by a PerkinElmer In Vivo Imaging System (IVIS) Spectrum and total bioluminescence was analyzed using Living Image software. Bioluminescence on day zero was compared to bioluminescence on the day of peak inflammation for each model. Longitudinal bioluminescence imaging was performed in EIU and EAU.ResultsIn the presence of luminol, intraocular inflammation generates detectable bioluminescence in three mouse models of uveitis. Peak bioluminescence in inflamed PMU eyes (1.46 × 105 photons/second [p/s]) was significantly increased over baseline (1.47 × 104 p/s, P = 0.01). Peak bioluminescence in inflamed EIU eyes (3.18 × 104 p/s) also was significantly increased over baseline (1.09 × 104 p/s, P = 0.04), and returned to near baseline levels by 48 hours. In EAU, there was a nonsignificant increase in bioluminescence at peak inflammation.ConclusionsIn vivo bioluminescence may be used as a noninvasive, quantitative measure of intraocular inflammation in animal models of uveitis. Primed mycobacterial uveitis and EIU are both acute models with robust anterior inflammation and demonstrated significant changes in bioluminescence corresponding with peak inflammation. Experimental autoimmune uveitis is a more indolent posterior uveitis and generated a more modest bioluminescent signal. In vivo imaging system bioluminescence is a nonlethal, quantifiable assay that can be used for monitoring inflammation in animal models of uveitis.

Osteopontin contributes to effective neutrophil recruitment, IL‐1β production and apoptosis in Aspergillus fumigatus keratitis

“…Mice were positioned on the IVIS warming stage in the infected eye lateral decubitus position with the ocular surface directly facing the camera sensor. In vivo imaging parameters were determined using standard optimization protocols . Mice were imaged using a field of view ‘C’ with medium binning for 1 min.…”

“…In vivo imaging parameters were determined using standard optimization protocols. 47,48 Mice were imaged using a field of view 'C' with medium binning for 1 min. Average bioluminescence at baseline and peak inflammation was compared by a paired Student's t-test using Prism-5 software.…”

Anterior segment optical coherence tomography: its application in clinical practice and experimental models of disease

“…The utility of OCT has also been compared to luminol‐based bioluminescence imaging in three rodent models of anterior uveitis . Here, the authors used SD‐OCT to assess anterior chamber cell counts and corneal oedema as robust signs of inflammation in the experimental models of anterior uveitis .…”

“…The utility of OCT has also been compared to luminol‐based bioluminescence imaging in three rodent models of anterior uveitis . Here, the authors used SD‐OCT to assess anterior chamber cell counts and corneal oedema as robust signs of inflammation in the experimental models of anterior uveitis . A recently‐developed method for automated 3‐D cell counting also demonstrates the potential for AS‐OCT to accurately detect and quantify cell populations within the whole anterior chamber, longitudinally up to 14 days post‐injury in a mouse model of uveitis .…”

Quantitative Characterization of Autoimmune Uveoretinitis in an Experimental Mouse Model

“…However, EAU was still difficult to quantify. 16,17 Optical coherence tomography (OCT) is a noninvasive in vivo imaging modality that provides structural information with resolution at micrometer level and can be performed repeatedly on living animals. Retinal OCT has been described for use in EAU previously to show the retinal changes.…”