# Immunopathogenesis of Adenovirus Keratitis

> **NIH NIH R01** · MASSACHUSETTS EYE AND EAR INFIRMARY · 2020 · $425,000

## Abstract

PROJECT SUMMARY
Epidemic keratoconjunctivitis (EKC), an important human eye infection, is caused by human adenovirus
species D (HAdV-D) types 8, 37, 53, 54, 56, and 64. The clinical manifestations of this infection include
severe membranous conjunctivitis and epithelial keratitis, followed by multifocal subepithelial (stromal)
corneal infiltrates that cause photophobia and reduced vision and may persist or recur for months to
years after the acute infection. We have shown previously that HAdV-D infection of human keratocytes
induces intracellular signaling and robust expression of neutrophil chemokines, and that physical
components of the adenovirus (viral capsid and DNA) initiate distinct cytokine responses in the corneal
stroma. However, characterization of responses to infection of other ocular surface cell types such as
corneal and conjunctival epithelial cells, is lacking,. The specific aims of this proposal are to test the
hypotheses that 1) HAdV-D37 infection of ocular surface cells induces cytokine signatures
distinguishable by cell type, 2) HAdV-D37 infection of ocular surface cells activates unique molecular
pattern recognition receptors distinguishable by cell type, and 3) cell type-specific pattern recognition
receptor activation by HAdV-D37 and intracellular trafficking of HAdV-D37 are interdependent. We will
apply intracellular cytokine staining with flow cytometry along with cytokine protein array analysis of the
various cell types on the human ocular surface to identify cell type-specific patterns of cytokine
expression upon HAdV-D37 infection. Because CXCL8 has been shown to increase subsequent
adenoviral entry by increasing the expression and apical localization of host cell receptors, we will
compare infectivity of corneal cells in the presence of cytokines expressed during infection of
conjunctival and immune cells from the ocular surface. We will apply Western blot, immuno-confocal
microscopy, ELISA, cytokine arrays, and siRNA knockdown, to determine the molecular pattern
receptors activated in human ocular surface and bone marrow derived cells by HAdV-D37 infection.
Finally, with methods that allow differentiation of intact virus versus capsid-free viral DNA and
endosomal versus cytoplasmic virus, with confocal microscopy, we will characterize viral trafficking and
its kinetics across ocular surface cell types, identify loci of interaction between viral DNA and specific
pattern recognition receptors, and evaluate the effect of pattern recognition receptor knockdown on
viral trafficking. The study of cytokine expression, pattern recognition receptor activation and signaling,
and intracellular trafficking across a broad spectrum of ocular surface cell types, will take us closer to
our goal of effective information-based therapies against HAdV eye infections. EKC is a common
affliction, and the proposed study addresses a major public health concern.

## Key facts

- **NIH application ID:** 9893874
- **Project number:** 5R01EY013124-19
- **Recipient organization:** MASSACHUSETTS EYE AND EAR INFIRMARY
- **Principal Investigator:** James Chodosh
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $425,000
- **Award type:** 5
- **Project period:** 2001-06-01 → 2022-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9893874

## Citation

> US National Institutes of Health, RePORTER application 9893874, Immunopathogenesis of Adenovirus Keratitis (5R01EY013124-19). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9893874. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*