TLR9 Ligand CpG-ODN Applied to the Injured Mouse Cornea Elicits Retinal Inflammation
Open accessAbstract
During bacterial and viral infections, unmethylated CpG-DNA released by proliferating and dying microbes is recognized by toll-like receptor (TLR) 9 in host cells, initiating innate immune responses. Many corneal infections occur secondary to epithelial breaches and represent a major cause of vision impairment and blindness globally. To mimic this clinical situation, we investigated mechanisms of TLR9 ligand-induced corneal inflammation in mice after epithelial debridement. Application of CpG oligodeoxynucleotides (ODNs) resulted in neutrophil and macrophage infiltration to the cornea and loss of transparency. By 6 hours after CpG-ODN administration, TLR9 mRNA was increased in the cornea and retina. In vivo clinical examination at 24 hours revealed inflammatory infiltrates in the vitreous and retina, which were confirmed ex vivo to be neutrophils and macrophages, along with activated resident microglia. CpG-ODN-induced intraocular inflammation was abrogated in TLR9(-/-) and macrophage-depleted mice. Bone marrow reconstitution of irradiated TLR9(-/-) mice with TLR9(+/+) bone marrow led to restored corneal inflammatory responses to CpG-ODN. Fluorescein isothiocyanate-CpG-ODN rapidly penetrated the cornea and ocular media to reach the retina, where it was present within CD68(+) retinal macrophages and microglia. These data show that topically applied CpG-ODN induces intraocular inflammation owing to TLR9 activation of monocyte-lineage cells. These novel findings indicate that microbial CpG-DNA released during bacterial and/or viral keratitis can cause widespread inflammation within the eye, including the retina.
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Topical application of CpG ODNs onto a scratched cornea leads to inflammation in eye tissues including retina, after CpG leakage into the anterior segment.
“Topical application of CpG ODNs onto a scratched cornea was shown by Chinnery et al. to lead to inflammation in eye tissues including retina, after CpG leakage into the anterior segment [88].”
From Risk mitigation of immunogenicity: a key to personalized retinal gene therapy (2021) · cites this paper as [88]
When CpG oligodeoxynucleotides were injected in C57BL/6 mice, neutrophilic infiltration was observed at the corneal stroma and a macrophage response induced by CpG ODNs was observed, confirming that naked DNA with CpG would induce ocular inflammation.
“When CpG oligodeoxynucleotides (ODNs) were injected in C57BL/6 mice, neutrophilic infiltration was observed at the corneal stroma and a macrophage response induced by CpG ODNs was observed, confirming that naked DNA with CpG would induce ocular inflammation [72].”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [72]
Depletion of CpG sequences in the viral genome helped reduce immune responses.
“The depletion of CpG sequences in the viral genome helped reduce immune responses (Figure 2) [72,95].”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [72]
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Research works printed but not linked: 3
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Ren, D., Fisson, S., Dalkara, D., & Ail, D. (2022) · Pharmaceutics · doi:10.3390/pharmaceutics14091973 · Cited by 52 · see citing works
Risk mitigation of immunogenicity: a key to personalized retinal gene therapy
Varin, J., Morival, C., Maillard, N., Adjali, O., & Cronin, T. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms222312818 · Cited by 19 · see citing works
- Authors
- Chinnery, H. R., McLenachan, S., Binz, N., Sun, Y., Forrester, J. V., Degli-Esposti, M. A., Pearlman, E., & McMenamin, P. G.
- Affiliations
- Lions Eye Institute (AU)
- Case Western Reserve University (US)
- University of Aberdeen (GB)
- Corresponding author
- McMenamin, Paul G.
- Venue
- The American Journal of Pathology
- Publisher
- Elsevier BV
- Published
- 2012-01
- Volume
- 180
- Issue
- 1
- Pages
- 209-220
- ISSN
- 0002-9440 (ISSN-L)
- PMC
- PMC3338340
- OpenAlex
- W2005818840
- Cited by
- 46
- References
- 49
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 2 · 2023: 2 · 2022: 1 · 2021: 1 · 2020: 5 · 2019: 2 · 2018: 3 · 2017: 3 · 2016: 4 · 2015: 7 · 2014: 6 · 2013: 5 · 2012: 2
- FWCI
- 1.84
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- American Journal Of Pathology (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- UWA Profiles and Research Repository (UWA) (submittedVersion, other-oa)
- Funders
- National Institutes of Health (R01 EY14362)
- National Health and Medical Research Council (572709)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Health Sciences › Medicine › Ophthalmology › Ocular Infections and Treatments
- Health Sciences › Medicine › Ophthalmology › Ocular Diseases and Behçet’s Syndrome
- Keywords
- TLR9, CpG oligodeoxynucleotides, keratitis, retinal inflammation, corneal epithelial debridement, neutrophil infiltration, macrophages, microglia, innate immunity, intraocular inflammation, topical immunotherapy
- MeSH terms
- Adjuvants, Immunologic, Animals, Cornea, Immunity, Innate, Keratitis, Lipopolysaccharides, Macrophages, Mice, Inbred BALB C, Mice, Inbred C57BL, Oligodeoxyribonucleotides, Retinitis, Neutrophil Infiltration, Toll-Like Receptor 4, Toll-Like Receptor 9, Mice, Corneal Injuries
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