Loss of Osteopontin Expression Reduces HSV-1-Induced Corneal Opacity

Abstract

Corneal opacity and neovascularization (NV) are often described as outcomes of severe herpes simplex virus type 1 (HSV-1) infection. The current study investigated the role of colony-stimulating factor 1 receptor (CSF1R) + cells and soluble factors in the progression of HSV-1-induced corneal NV and opacity. METHODS. MaFIA mice were infected with 500 plaque-forming units of HSV-1 in the cornea following scarification. From day 10 to day 13 post-infection (pi), mice were treated with 40 μg/day of AP20187 (macrophage ablation) or vehicle intraperitoneally. For osteopontin (OPN) neutralization experiments, C57BL/6 mice were infected as above and treated with 2 μg of goat anti-mouse OPN or isotypic control IgG subconjunctivally every 2 days from day 4 to day 12 pi. Mice were euthanized on day 14 pi, and tissue was processed for immunohistochemistry to quantify NV and opacity by confocal microscopy and absorbance or detection of pro-and anti-angiogenic and inflammatory factors and cells by suspension array analysis and flow cytometry, respectively. RESULTS. In the absence of CSF1R + cells, HSV-1-induced blood and lymphatic vessel growth was muted. These results correlated with a loss in fibroblast growth factor type 2 (FGF-2) and an increase in OPN expression in the infected cornea. However, a reduction in OPN expression in mice did not alter corneal NV but significantly reduced opacity. CONCLUSIONS. Our data suggest that CSF1R + cell depletion results in a significant reduction in HSV-1-induced corneal NV that correlates with the loss of FGF-2 expression. A reduction in OPN expression was aligned with a significant drop in opacity associated with reduced corneal collagen disruption.

HSV-1 0∆NLS vaccine elicits a robust B lymphocyte response and preserves vision without HSV-1 glycoprotein M or thymidine kinase recognition

“…In mice, acute ocular HSV-1 infection often results in cornea pathology including denervation 39 , 40 , opacity as a result of collagen fiber disruption 41 , inflammatory hypoxia 42 , local changes in substance P levels 43 , and products of infiltrating leukocytes 44 , hemangiogenesis 45 , lymphangiogenesis 46 , and edema 47 leading to blindness or significant vision loss 48 . In previous work, we found a clear association with vaccine efficacy and preservation of the visual axis in terms of architecture, cornea integrity, and vision performance 26 .…”

Tripartite-Motif 21 (TRIM21) Deficiency Results in a Modest Loss of Herpes Simplex Virus (HSV)-1 Surveillance in the Trigeminal Ganglia Following Cornea Infection

“…All animals were housed in a specific pathogen-free vivarium at the Dean McGee Eye Institute on the University of Oklahoma Health Sciences Center campus, under the approved institutional animal use and care committee protocol, 19-060-ACHIX. At the indicated time, mice were exsanguinated, as previously described [ 33 ].…”

“…The opacity of the cornea was determined at 0, 7, and 30 days pi, following the removal of the cornea button from exsanguinated mice and assaying the tissue for absorbance of 500 nm using FLUOstar Omega plate reader (BMG Labtech, Offenburg, Germany), as described previously [ 33 ]. Following the measurement, the cornea was fixed using 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, USA) for 30 min and washed in PBS containing 1% Triton X-100 (Sigma Aldrich).…”

Lymphatic Vessel Regression and Its Therapeutic Applications: Learning From Principles of Blood Vessel Regression

“…Inflammation-induced angiogenesis has been studied in a variety of contexts, including herpes simplex virus 1–induced corneal neovascularization, helicobacter pylori infection, cancer, and inflammatory bowel disease ( Sajib et al, 2018 ; Filiberti et al, 2020 ). Sajib et al (2018) described the intimate relationship between inflammation and angiogenesis, citing the dual role inflammatory markers such as IL-8 and cyclooxygenase-2 may play in both inflammation and promoting angiogenesis.…”