July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Mast cells initiate early recruitment of neutrophils following corneal injury
Author Affiliations & Notes
  • Sharad Mittal
    Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Srikant Sahu
    Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Mingshun Li
    Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Afsaneh Amouzegar
    Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • William Foulsham
    Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Sunil Chauhan
    Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 3316. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Sharad Mittal, Srikant Sahu, Mingshun Li, Afsaneh Amouzegar, William Foulsham, Sunil Chauhan; Mast cells initiate early recruitment of neutrophils following corneal injury. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3316.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : Injury-induced innate immune responses, primarily the recruitment of activated neutrophils to the eye, lead to deleterious inflammatory tissue damage. This study investigated the contribution of resident mast cells to the initiation of neutrophil recruitment following corneal injury.

Methods : Corneal injury was created by removal of the epithelium and anterior stroma using a hand-held motor brush. Corneas and conjunctivae were harvested to investigate frequencies of CD45+ inflammatory cells, CD11b+Ly6G+ neutrophils and c-Kit+FcεR1+ mast cells by flow cytometry. Mast cell-activating factors IL1β and IL33 were evaluated using real-time PCR and ELISA. In vitro, mast cells were stimulated with mechanically damaged corneas in the presence or absence of IL1β or IL33 neutralizing antibodies and assessed for the expression of β-hexosaminidase, CXCL1 and CXCL2 using real-time PCR and ELISA. Cromolyn (2% in PBS) or IL1β neutralizing antibodies (1 mg/ml) were administered locally for mast cell inhibition in vivo.

Results : Flow cytometry analysis demonstrated early neutrophil infiltration to the cornea within 1 hour of injury (3±0.6%, p<0.05), and this infiltration increased progressively over the first 12 hours following injury (12±1%, p<0.001) compared to normal cornea (1±0.2%). Frequencies of activated mast cells at the ocular surface were significantly (3-fold) increased until 6 hours post-injury (0.9±0.3%, p<0.05) and then declined to baseline levels by 12 hours (0.3±0.2%). Mast cells constitutively expressed CXCL2. Mast cells cultured with injured corneas secreted 2.5-fold higher levels of CXCL2 compared to mast cells cultured alone (p<0.01). Corneal injury resulted in significantly increased levels of IL1β and IL33 at the ocular surface compared to normal corneas (p<0.05). IL1β neutralization in injured cornea-mast cell cocultures significantly inhibited mast cell activation and CXCL2 secretion compared to isotype-treated control cocultures (p<0.05). Moreover, in vivo topical blockade of IL1β significantly inhibited mast cell activation, expression of CXCL2 (p<0.05) and limited the early infiltration of neutrophils to the ocular surface compared to control isotype-treated injured corneas (1.9±0.3% vs. 9.8±2%, p<0.01).

Conclusions : Our data suggest that corneal injury activates mast cells via IL1β and amplifies mast cell secretion of CXCL2, which is associated with early neutrophil recruitment to the ocular surface.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×