July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Secreted Ly-6/uPAR Related Protein-1 (SLURP1) modulates inflammation by moderating epithelial cells response to inflammatory agents
Author Affiliations & Notes
  • Sudha Swamynathan
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Gregory Campbell
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Anil Tiwari
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • John Sebastian Gnalian
    Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Shivalingappa K Swamynathan
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Footnotes
    Commercial Relationships   Sudha Swamynathan, US 9,731,014 B2 (P); Gregory Campbell, None; Anil Tiwari, None; John Gnalian, None; Shivalingappa Swamynathan, US 9,731,014 B2 (P)
  • Footnotes
    Support  NIH Grant R01EY026533 (SKS), NEI Core Grant P30 EY08098, unrestricted grants from Research to Prevent Blindness and the Eye and Ear Foundation of Pittsburgh.6533
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 915. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Sudha Swamynathan, Gregory Campbell, Anil Tiwari, John Sebastian Gnalian, Shivalingappa K Swamynathan; Secreted Ly-6/uPAR Related Protein-1 (SLURP1) modulates inflammation by moderating epithelial cells response to inflammatory agents. Invest. Ophthalmol. Vis. Sci. 2019;60(9):915. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Previously we demonstrated that the Secreted Ly-6/uPAR Related Protein-1 (SLURP1), abundantly expressed in the cornea and secreted into the tear film, serves as an anti-angiogenic and anti-inflammatory molecule. Here we have studied the effect of SLURP1 on the inflammatory response of Human Corneal Limbal Epithelial (HCLE) cells to tumor necrosis factor-α (TNF-α).

Methods : HCLE cells were stably transfected to express SLURP1 and two different clones were selected for the present studies. The effect of SLURP1 on TNF-α-induced expression of (i) IL-8 was quantified by QPCR and ELISA, (ii) CXCL1, CXCL2 and IL-1b by QPCR, in un-transfected control and SLURP1-expressing HCLE cells. The effect of SLURP1 on TNF-α-induced NF-kB nuclear translocation was studied by immunofluorescent staining and immunoblots. The effect of SLURP1 on TNF-α-induced oxidative stress was examined by immunoblots and immunofluorescent staining with anti-4-Hydroxynonenal (4-HNE) antibody.

Results : QPCR revealed a significant decrease in TNF-α-induced upregulation of (i) IL-8 from 7.4- to 2.9- and 2.1 fold, (ii) IL-1b from 4.9- to 3.9- and 2.9-fold, (iii) CXCL1 from 9- to 3.3- and 5.5-fold, and (iv) CXCL2 from 4.8- to 2.1- and 2.8-fold respectively, in two different SLURP1-expressing HCLE clones compared with the WT cells. ELISAs revealed a concomitant decrease in IL-8 levels in cell culture supernatants from 789 pg/ml in the WT, to 503 and 352 pg/ml in the SLURP1-expressing HCLE clones. Immunoblots and immunofluorescent staining with anti-4-HNE antibodies revealed lower TNF-α-induced oxidative stress in SLURP1-expressing HCLE cells compared with the WT. Furthermore, TNF-α-activated nuclear translocation of NF-kB was suppressed in SLURP1-expressing HCLE cells.

Conclusions : Collectively, these results demonstrate that SLURP1 (i) suppresses the TNF-α-induced upregulation of IL-8, IL-1b, CXCL-1 and CXCL-2 in HCLE cells by blocking NF-kB nuclear translocation, and (ii) safeguards the HCLE cells from TNF-α-induced oxidative stress.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

×
×

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.

×