Ephrin-B2 is differentially expressed in the intestinal epithelium in Crohn's disease and contributes to accelerated epithelial wound healing in vitro.
OBJECTIVE
Eph receptor tyrosine kinases and their membrane bound receptor-like ligands, the ephrins, represent a bi-directional cell-cell contact signaling system that directs epithelial movements in development. The meaning of this system in the adult human gut is unknown. We investigated the Eph/ephrin mRNA expression in the intestinal epithelium of healthy controls and patients with inflammatory bowel disease (IBD).


METHODS
mRNA expression profiles of all Eph/ephrin family members in normal small intestine and colon were established by real-time RT-PCR. In addition, differential expression in IBD was investigated by cDNA array technology, and validated by both real-time RT-PCR and immunohistochemistry. Potential effects of enhanced EphB/ephrin-B signaling were analyzed in an in vitro IEC-6 cell scratch wound model.


RESULTS
Human adult intestinal mucosa exhibits a complex pattern of Eph receptors and ephrins. Beside the known prominent co-expression of EphA2 and ephrinA1, we found abundantly co-expressed EphB2 and ephrin-B1/2. Interestingly, cDNA array data, validated by real-time PCR and immunohistochemistry, showed upregulation of ephrin-B2 in both perilesional and lesional intestinal epithelial cells of IBD patients, suggesting a role in epithelial homeostasis. Stimulation of ephrin-B signaling in ephrin-B1/2 expressing rat IEC-6-cells with recombinant EphB1-Fc resulted in a significant dose-dependent acceleration of wound closure. Furthermore, fluorescence microscopy showed that EphB1-Fc induced coordinated migration of wound edge cells is associated with enhanced formation of lamellipodial protrusions into the wound, increased actin stress fiber assembly and production of laminin at the wound edge.


CONCLUSIONS
EphB/ephrin-B signaling might represent a novel protective mechanism that promotes intestinal epithelial wound healing, with potential impact on epithelial restitution in IBD.
Department of Dermatology, University of Regensburg, D-93042 Regensburg, Germany.
Christian
C
Hafner
Stefanie
S
Meyer
Thomas
T
Langmann
Gerd
G
Schmitz
Frauke
F
Bataille
Ilja
I
Hagen
Bernd
B
Becker
Alexander
A
Roesch
Gerhard
G
Rogler
Michael
M
Landthaler
Thomas
T
Vogt
D003424:Crohn Disease / Q000503:physiopathology*; D036388:Ephrin-B2 / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D007413:Intestinal Mucosa / Q000473:pathology* / Q000502:physiology / Q000503:physiopathology; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014945:Wound Healing / Q000235:genetics*
D036388:Ephrin-B2; D012313:RNA