June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Immunological characterization of chemical burn-induced ocular surface pannus in humans, rabbits and mice after limbal stem cell deficiency.
Author Affiliations & Notes
  • Vivek Singh
    Center for Ocular Regeneration (CORE), LV Prasad Eye Institute ( Hyderabad Eye Research Foundation), Hyderabad, Telangana, India
    Tej Kohli Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Harsha Agarwal
    Center for Ocular Regeneration (CORE), LV Prasad Eye Institute ( Hyderabad Eye Research Foundation), Hyderabad, Telangana, India
    Tej Kohli Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Abhinav Reddy Kethiri
    Center for Ocular Regeneration (CORE), LV Prasad Eye Institute ( Hyderabad Eye Research Foundation), Hyderabad, Telangana, India
    Tej Kohli Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Mukesh Damala
    Center for Ocular Regeneration (CORE), LV Prasad Eye Institute ( Hyderabad Eye Research Foundation), Hyderabad, Telangana, India
    Tej Kohli Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Sayan Basu
    Center for Ocular Regeneration (CORE), LV Prasad Eye Institute ( Hyderabad Eye Research Foundation), Hyderabad, Telangana, India
    Tej Kohli Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Virender Singh Sangwan
    Center for Ocular Regeneration (CORE), LV Prasad Eye Institute ( Hyderabad Eye Research Foundation), Hyderabad, Telangana, India
    Tej Kohli Cornea Institute, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Footnotes
    Commercial Relationships   Vivek Singh, None; Harsha Agarwal, None; Abhinav Reddy Kethiri, None; Mukesh Damala, None; Sayan Basu, None; Virender Sangwan, None
  • Footnotes
    Support  Research Grant (Young Scientists) by SCIENCE & ENGINEERING RESEARCH BOARD (SERB)- Registration No.SERB/LS-599/2013 ( 2014-2017).
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 1423. doi:
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      Vivek Singh, Harsha Agarwal, Abhinav Reddy Kethiri, Mukesh Damala, Sayan Basu, Virender Singh Sangwan; Immunological characterization of chemical burn-induced ocular surface pannus in humans, rabbits and mice after limbal stem cell deficiency.. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1423.

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

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Abstract

Purpose : This study evaluated the validity of using rabbit and mouse animal models of limbal stem cell deficiency (LSCD). We also evaluated the biology of corneal damage in ocular burns by comparing the histological and immunological characteristics of alkali burn-induced chronic ocular surface pannus in humans, rabbit and mouse eyes.

Methods : Human samples (n=12) were obtained from eyes undergoing excision of the pathological pannus during limbal stem cell transplantation. The animal models of unilateral chronic ocular alkali burns were created by a single application of sodium hydroxide (0.1N) followed by 8 to 12 weeks before sacrificing the adult New Zealand white rabbits (n=20) and C57/BL6 black mice (n=12). Paraffin fixed tissue sections were stained with Hematoxylin-Eosin (H&E) and Periodic acid–Schiff (PAS) to study the histopathological features. Immunohistochemistry was performed to identify and quantify immune infiltrates by detecting the expression of T- cells (CD3/5), B-cells (CD20) and plasma B-cell (CD138). The signaling pathways involving CD40 were also investigated.

Results : Granulomatous tissue with inflammation was noted in all human samples with epithelial hyperplasia, neo-vascularization, surface ulceration and active fibrosis. The common histological features included presence of goblet cells on the corneal surface (75%), multinucleated giant cells in the underlying stroma (50%) and keratinization of the epithelium (33.3%), thus confirming the clinical diagnosis of LSCD. Similar histopathological features were observed in excised corneas of both rabbits and mice (p>0.05). Lymphocytic infiltrates in the human pannus were found to be positive for CD3 (22.3±2.7%), CD5 (13.1±1.8%), CD20 (5.36±1.1%) and CD138 (6.3±2.1%). We also obsereved CD40 poitive cells in these pannus.

Conclusions : The findings of the study suggest that both the rabbit and the mouse models of ocular alkali burns closely mimic the human condition. The presence of inflammatory cells many months after the acute burn may potentially lead to limbal or corneal transplant rejection in the future. Therefore, identification and regulation of the local inflammation prior to limbal transplantation needs to be contemplated in human eyes.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

 

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