Abstract Details

To investigate the expression pattern of Ephrin receptors and their ligand on human microglial cell line and test the effect of Eph/sEphB4-HSA interaction  on microglial biology.

sEphB4-HSA is a fully human fusion protein of soluble EphB4 extracellular domain fused with albumin. sEphB4, is designated as an antagonist of EphB4-Epherin binding and activation, which can inhibit its signaling pathway

We use real time quantitative RT-PCR to determine the expressions of different Eph receptors and their ligand ephrins as follow: EphB1, EphB2, EphB3, EphB4, EphB6, ephrin-B1, ephrin-B2 and ephrin-B6. Furthermore, we used sEphB4-HAS to treat CHME-5 to evaluate the effects on cell proliferation.

Quantitative RT-PCR showed CHME-5 cell express all Eph receptors and ephrins except EphB2 (which we did not detect positive signal in 40 cycles). No effect on microglia survival was observed after 24 hours exposure to 20ug/ml sEphB4-HAS or 1ug/ml LPS alone. However, the combination of sEphB4-HAS and 1ug/ml LPS demonstrated significant reduction of viability. Compared to LPS alone (p<0.05) and sEphB4-HAS alone (p<0.005. Similar results were observed with 10 ug/ml of sEphB4-HAS

sEphB4-HAS may have a synergistic effect with LPS on human microglia cell line viability and EphB4-Epherin pathways. Work in progress to elucidate the effect of sEphB4-HAS on human microglia survival, proliferation and activation.