Purchase this article with an account.
Birgit Regenfuss, Deniz Hos, Felix Bock, Jasmine Onderka, Sharmila Masli, Claus Cursiefen; TRAIL Influences Human Lymphatic Microvascular Endothelial Cell (HMVECs) Proliferation and Lymphangiogenesis. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2398.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Several (lymph-) angiogenic candidate genes were found to be differentially expressed between C57BL/6 and BALB/c or FVB mice with significantly more resting limbal vasculature in the former strain than the latter two. One of the candidate genes, TRAIL/Tnfsf10 was analyzed in greater detail to obtain insights into its effect on lymphatic endothelial cells.
Corneal RNA was isolated and cDNA synthesis was carried out with SuperScriptTM III First-Strand Synthesis SuperMix. The cDNA was used for real-time PCR analyses. Lymphatic endothelial cell proliferation was analysed using BrdU incorporation into newly synthesized DNA. Surface expression of a corresponding death receptor of TRAIL (TRAIL-R2/DR5) was assessed by FACS analysis and immunocytochemistry.
Expression of TRAIL mRNA was increased 8.2-fold in BALB/c and 6.6-fold in FVB corneas compared to the C57BL/6 control corneas. This expression pattern corresponded with the decreased limbal vasculature in BALB/c and FVB compared to C57BL/6. Treatment of HMVECs with different concentrations of recombinant TRAIL in endothelial cell basal medium (containing 0.5 % FBS) resulted in a small, but significant inhibition of proliferation at a concentration of 1 ng/ml TRAIL. Flow cytometric analysis of surface expression of TRAIL-R2/DR5 (a corresponding death receptor of TRAIL) on HMVECs confirmed the presence of a ligand for TRAIL with an increased expression detectable after TNFα treatment (1 ng/ml and 100 ng/ml).
Corneal expression of TRAIL may play an inhibitory role in lymphatic endothelial cell biology. Further studies are needed to clarify the detailed function of TRAIL in lymphangiogenesis.
This PDF is available to Subscribers Only