Model Systems for Studying Mechanisms of Ocular Toxoplasmosis

Smith, Justine R.; Ashander, Liam M.; Ma, Yuefang; Rochet, E; Furtado, João M.

doi:10.1007/978-1-4939-9857-9_17

Cited by 10 publications

(9 citation statements)

References 47 publications

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“…In the animal model, we induced acute ocular toxoplasmosis by intraocular injection of tachyzoites. It has been reported that the advantage of this method is that it can breach ocular immune privilege and rapidly induces vitritis and retinitis before the onset of systemic disease in mice ( Dunay et al., 2018 ; Smith et al., 2020 ). In terms of treatment, the general principle of the therapy is to inhibit parasites proliferation, immediately control the inflammatory response, and mitigate tissue damage to protect retina during acute infection ( Casoy et al., 2020 ).…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, we also selected this method to evaluate the therapeutic effect of LBH589 instead of intraperitoneal injection or oral gavage. In order to minimize the secondary damage to the mouse’s eyes and set aside time for T. gondii infection, mice were injected with LBH589 into the eyes 12 h after infection with T. gondii , and performed clinical evaluation on the 7th day when the ocular symptoms of the mice were most obvious ( LaGrow et al., 2017 ; Lafreniere et al., 2019 ; Kishimoto et al., 2019 ; Smith et al., 2020 ). Here, we first adopted the ophthalmic imaging diagnostic technology, OCT examination, which can directly non-invasive assessment of intraocular lesion in living mice ( Cunningham et al., 2022 ).…”

Section: Discussionmentioning

confidence: 99%

“…After the pupils of the anesthetized mice were dilated, the eyes were observed under a surgical microscope and a tiny incision was made 1 mm behind the limbus. Free tachyzoites (4000/2 μl) were injected through a Hamilton syringe into the vitreous, making sure that the tip does not damage the lens or retina ( Smith et al., 2020 ). At 12 h after infection, the treatment group was injected with different doses of LBH589 (1, 2, 3 ng/μl) via intravitreal injection, and the infection group was injected with an equal volume of PBS.…”

Section: Methodsmentioning

confidence: 99%

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In vitro and in vivo anti−Toxoplasma activities of HDAC inhibitor Panobinostat on experimental acute ocular toxoplasmosis

Zhang¹,

Li²,

Huang³

et al. 2022

Front. Cell. Infect. Microbiol.

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Ocular toxoplasmosis (OT) is retinochoroiditis caused by Toxoplasma gondii infection, which poses a huge threat to vision. However, most traditional oral drugs for this disease have multiple side effects and have difficulty crossing the blood-retinal barrier, so the new alternative strategy is required to be developed urgently. Histone deacetylases (HDAC) inhibitors, initially applied to cancer, have attracted considerable attention as potential anti-Toxoplasma gondii drugs. Here, the efficacy of a novel HDAC inhibitor, Panobinostat (LBH589), against T. gondii has been investigated. In vitro, LBH589 inhibited the proliferation and activity of T. gondii in a dose-dependent manner with low toxicity to retinal pigment epithelial (RPE) cells. In vivo, optical coherence tomography (OCT) examination and histopathological studies showed that the inflammatory cell infiltration and the damage to retinal architecture were drastically reduced in C57BL/6 mice upon treatment with intravitreal injection of LBH589. Furthermore, we have found the mRNA expression levels of inflammatory cytokines were significantly decreased in LBH589–treated group. Collectively, our study demonstrates that LBH589 holds great promise as a preclinical candidate for control and cure of ocular toxoplasmosis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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In vitro and in vivo anti−Toxoplasma activities of HDAC inhibitor Panobinostat on experimental acute ocular toxoplasmosis

Zhang¹,

Li²,

Huang³

et al. 2022

Front. Cell. Infect. Microbiol.

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“…Human retinal pigment epithelial cells were isolated from human cadaveric eyes (5 male and 3 females, aged 50 to 75 years at death), following a method we have previously published [15]. Death-to-processing time ranged from 15 to 42 h.…”

Section: Human Retinal Cellsmentioning

confidence: 99%

Infection of Human Retinal Pigment Epithelial Cells with Dengue Virus Strains Isolated during Outbreaks in Singapore

et al. 2022

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Prevalence of dengue retinopathy varies across epidemics, with the disease linked to circulation of dengue virus serotype 1 (DENV-1). The retinal pigment epithelium has been implicated in the pathology. We investigated infectivity, molecular response, and barrier function of epithelial cells inoculated with DENV strains from different outbreaks in Singapore. Monolayers of human retinal pigment epithelial cells (multiple primary cell isolates and the ARPE-19 cell line) were inoculated with six DENV strains, at multiplicity of infection of 10; uninfected and recombinant strain-infected controls were included where relevant. Infectivity and cell response were assessed primarily by RT-qPCR on total cellular RNA, and barrier function was evaluated as electrical resistance across monolayers. Higher viral RNA loads were measured in human retinal pigment epithelial cells infected with DENV-1 strains from the 2005 Singapore epidemic, when retinopathy was prevalent, versus DENV-1 strains from the 2007 Singapore epidemic, when retinopathy was not observed. Type I interferon (IFN) transcripts (IFN-β and multiple IFN-stimulated genes) were up-regulated, and impact on barrier function was more pronounced, for cells infected with DENV-1 strains from the 2005 versus the 2007 Singapore epidemics. Aside from serotype, strain of DENV may determine the potential to induce retinal pathology. Identification of molecular markers of disease-associated DENV strains may provide insights into the pathogenesis of dengue retinopathy.

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“…Five retinal endothelial cell isolates were prepared separately from human cadaver donor eyes, using the positive selection procedure that we have described comprehensively in Methods in Molecular Biology , 12 and used in diverse, published research. 13 – 16 These individual isolates were stored in liquid nitrogen prior to transcriptomic profiling.…”

Section: Introductionmentioning

confidence: 99%

Transcriptomic Responses of Human Retinal Vascular Endothelial Cells to Inflammatory Cytokines

Ryan

Ashander

et al. 2022

Trans. Vis. Sci. Tech.

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Purpose Molecular profiling of human retinal endothelial cells provides opportunities to understand the roles of this cell population in maintenance of the blood-ocular barrier, and its involvements in diverse retinal vasculopathies. We aimed to generate a transcriptome of human retinal endothelial cells in the unstimulated state, and following treatment with inflammatory cytokines linked to cell dysfunction. Methods Endothelial cells were isolated from retinae of five human cadaveric donors, and treated for 60 minutes and 24 hours with interleukin-1β or tumor necrosis factor-α, or exposed to medium alone for the same intervals. Expression of intercellular adhesion molecule-1 was measured by RT-qPCR to confirm cytokine-induced activation of the cells. RNA was sequenced on the Illumina NovaSeq 6000 platform. Reads were aligned to the human GRCh38 genome, and reads that aligned to Ensembl-annotated genes were counted. Quality control of sequencing was performed with FastQC, and sequences were classified by Kraken. Results A human retinal endothelial cell RNA-sequencing dataset with mean of 99% reads aligned to the human genome was produced as raw RNA sequence data (FASTQ files) and processed read data (XLSX files). Multidimensional scaling analysis showed a strong donor effect, which was readily controlled by ComBat. Conclusions Our dataset may be useful for human retinal endothelial cell transcriptomic assemblies, functional gene annotating and/or gene expression and enrichment analyses, as well as cross-dataset harmonization. Translational Relevance The molecular profile of the human retinal endothelium is a source of candidate biologic targets for retinal vasculopathies.

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Model Systems for Studying Mechanisms of Ocular Toxoplasmosis

Cited by 10 publications

References 47 publications

In vitro and in vivo anti−Toxoplasma activities of HDAC inhibitor Panobinostat on experimental acute ocular toxoplasmosis

In vitro and in vivo anti−Toxoplasma activities of HDAC inhibitor Panobinostat on experimental acute ocular toxoplasmosis

Infection of Human Retinal Pigment Epithelial Cells with Dengue Virus Strains Isolated during Outbreaks in Singapore

Transcriptomic Responses of Human Retinal Vascular Endothelial Cells to Inflammatory Cytokines

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