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Fgl2 induction in porcine endothelial cells through xenogeneic CD40-CD40L interaction

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Authors
Cho, Bumrae; Choi, Inho; Hurh, Sunghoon; Lee, Eun Mi; Ro, Han; Oh, Kook-Hwan; Yang, Jaeseok; Ahn, Curie; Kim, Jae Young; Hwang, Jong-ik; Kim, Hwa Jung
Issue Date
2009-10
Publisher
WILEY-BLACKWELL PUBLISHING, INC
Citation
XENOTRANSPLANTATION; Vol.16 5; 371-371
Abstract
Background: Fibrinogen-like protein 2 (fgl2) is a direct prothrombinase that
generates thrombin from prothrombin in the absence of a classical
prothrombinase complex. Fgl2 is known to be involved in experimental
xenograft rejection by mediating coagulation, fibrin deposition and microthrombus
formation, leading to classical pathological changes of acute
vascular rejection. We tried to investigate whether interaction between CD40
on porcine endothelial cells (PECs) and CD40L on human cells mediates
induction of fgl2 on PECs.
Methods: Fgl2 mRNA expression was assessed using reverse-transcriptase
polymerase chain reaction (RT-PCR). Protein expression and enzymatic
activity of fgl2 in PECs were assessed using western blot and fgl2 thrombin
generation assay, respectively. Both porcine fgl2 and CD40 knockdowned
stable cell lines were established using sifgl2 and siCD40 RNAi system.
Results: Fgl2 mRNA expression was induced in PECs at 30 min after PECs
had been cultured with Jurkat D1.1 and its expression was suppressed by
anti-CD40L antagonistic antibody treatment. Protein expression of fgl2 in
PECs was up-regulated at 4 h after treatment of anti-CD40 agonistic
antibody as well as TNF-a.
The enzymatic activity of fgl2 on PECs was increased by about
twofold at 6 h after stimulation of anti-CD40 agonistic antibody. Stable
cell lines for either sifgl2 or siCD40 were established. The enzymatic
activity of fgl2 was diminished by both sifgl2 and siCD40, but not
affected by a control siEGFP.
Conclusion: Fgl2 can be up-regulated and mediate coagulation response in
activated PECs through xenogeneic CD40-CD40L interaction. Therefore, it
might be helpful for the successful xenotransplantation to control fgl2
induction through development of porcine CD40 knockout pigs or antagonistic
anti-CD40 treatment.
ISSN
0908-665X
Language
English
URI
http://hdl.handle.net/10371/77600
DOI
https://doi.org/10.1111/j.1399-3089.2009.00539_3.x
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College of Medicine/School of Medicine (의과대학/대학원)Internal Medicine (내과학전공)Journal Papers (저널논문_내과학전공)
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