Regulation of the receptor for TNFalpha, TNFR1, in human conjunctival epithelial cells.

Neal Barney // Publications // Sep 01 2008

PubMed ID: 18487372

Author(s): Cook EB, Stahl JL, Graziano FM, Barney NP. Regulation of the receptor for TNFalpha, TNFR1, in human conjunctival epithelial cells. Invest Ophthalmol Vis Sci. 2008 Sep;49(9):3992-8. doi: 10.1167/iovs.08-1873. Epub 2008 May 16. PMID 18487372

Journal: Investigative Ophthalmology & Visual Science, Volume 49, Issue 9, Sep 2008

PURPOSE Previous studies demonstrated that mast cell-derived TNFalpha stimulation is critical to the upregulation of intercellular adhesion molecule (ICAM)-1 on human conjunctival epithelial cells (HCECs), which is an important feature of ocular allergic inflammation. Shedding of TNFR1 by TNFalpha-converting enzyme (TACE) is a primary mechanism for the regulation of TNFalpha-mediated events. This process has not been examined in HCECs. In this study, the authors examined the regulation of TNFR1 expression and shedding by TACE on primary HCECs and the IOBA-NHC conjunctival epithelial cell line.

METHODS Primary human conjunctival mast cells and epithelial cells were obtained from cadaveric conjunctival tissue. HCECs were incubated with and without activators (IgE-activated mast cell supernates, phorbol myristate acetate [PMA; to activate TACE], TNFalpha, and IFNgamma [to upregulate TNFR1]) for 24 hours. Pretreatment with the TACE inhibitor TAPI-2 was used to inhibit shedding of TNFR1. Supernates collected from the incubations were analyzed with ELISA for soluble TNFR1 (sTNFR1). With the use of flow cytometry, cells were harvested from these experiments for analysis of TNFR1 and ICAM-1 receptor expression.

RESULTS IgE-activated conjunctival mast cell supernates upregulated the expression of TNFR1. TAPI-2 inhibited the PMA-induced release of sTNFR1 receptor and enhanced the surface expression of TNFR1 in HCECs in a dose-dependent manner. Upregulation of TNFR1 expression by priming with TAPI-2 and IFNgamma resulted in enhanced ICAM-1 expression in response to TNFalpha stimulation (significant change in the slope of the dose-response curve).

CONCLUSIONS These results demonstrate that TACE promotes TNFR1 shedding in HCECs and that TNFR1 expression may be a more significant target than TNFalpha for intervention in ocular inflammation.