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S. Groman-Lupa, L. Vizuet-Garcia, G. A. Luna-Baca, A. Robles-Contreras, E. Rivera-Mendez, J. Serafin, I. Estrada-Garcia, S. Estrada-Parra S, M. Perez-Tapia, M. C. Jimenez-Martinez; Dyalizable Leukocyte Extracts and TLR Agonist Effects on VEGF Secretion by Human Limbal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):415.
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© ARVO (1962-2015); The Authors (2016-present)
To study the effect of Dializable Leukocyte Extracts (DLE) and TLR agonists on VEGF secretion by Human Limbal Epithelial Cells (HuLEC).
HuLEC were obtained from cadaveric corneal-scleral rings and cultured with KSFM until confluence. Then, HuLEC were stimulated at optimal concentrations with specific TLR1-9 agonists (TLRa): Pam3CSK4, HKLM, Poly I:C, LPS, Flagellin, FSL-1, Imiquimod, ssRNA40 and ODN2006, PMA/Ionomicyne or DLE. Supernatants were collected at different times and analyzed to determine VEGF concentrations by ELISA.
VEGF concentrations at 24h: Non Treated HuLEC 315.1pg/ml, PMA/Ionomicyne 395pg/ml; Pam3CSK4 treated HuLEC 262.2pg/mL, HKLM treated HuLEC 415.8pg/mL, Poly I:C treated HuLEC, 430.3pg/mL, LPS treated HuLEC 307.2pg/mL, Flagellin treated HuLEC 340.0pg/mL, FSL-1 treated cells 343.6pg/mL, Imiquimod treated HulEC 433.6pg/mL, ssRNA40 treated cells 355.0pg/mL, ODN2006 treated cells 322.4pg/mL, DLE 0.5µg/mL treated HuLEC 755.4pg/mL, DLE 5µg/mL treated HuLEC 1310pg/mL.
DLE induced up-regulation of VEGF secretion by HuLEC. These results suggest that DLE stimulation could be used as an in vitro model to study molecules related to neovascularization in cells derived from ocular surface. Stimulation trough TLRa (HKLM, Poly I:C and Imiquimod) could be associated with innate pathways of neovascularization in HuLEC. Additional studies are needed to assess this question.
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