Caja Sergio, Myrsky Essi, Korponay-Szabo Ilma R, Nadalutti Cristina, Sulic Ana-Marija, Lavric Miha, Sblattero Daniele, Marzari Roberto, Collighan Russell, Mongeot Alexandre, Griffin Martin, Mäki Markku, Kaukinen Katri, Lindfors Katri
Paediatric Research Centre, Medical School, University of Tampere, Tampere, Finland.
Scand J Gastroenterol. 2010 Apr;45(4):421-7. doi: 10.3109/00365520903540822.
Earlier work has demonstrated that serum autoantibodies from coeliac patients targeted against transglutaminase 2 (TG2) inhibit in vitro angiogenesis. The aim of this study was to establish whether coeliac patient-derived monoclonal TG2-targeted antibodies produced by recombination technology exert similar anti-angiogenic effects to serum-derived coeliac autoantibodies. In addition, we studied whether the monoclonal patient autoantibodies modulate endothelial cell TG2 activity and whether such modulation is related to the anti-angiogenic effects.
The influence of coeliac patient-derived monoclonal TG2-targeted antibodies on endothelial cell tubule formation was studied using a three-dimensional angiogenic cell culture model. Endothelial cell TG2 enzymatic activity was determined by means of a live-cell enzyme-linked immunosorbent assay.
Coeliac patient-derived monoclonal TG2-targeted antibodies produced by recombination technology inhibited endothelial tubule formation and enhanced the crosslinking activity of TG2. When this enzymatic activity was inhibited using site-directed irreversible TG2 inhibitors in the presence of autoantibodies, in vitro angiogenesis reverted to the control level.
Since we found a significant negative correlation between endothelial cell angiogenesis and TG2 activity, we suggest that the anti-angiogenic effects of coeliac patient-derived TG2-targeted autoantibodies are exerted by enhanced enzymatic activity of TG2.
早期研究表明，腹腔疾病患者针对转谷氨酰胺酶2（TG2）的血清自身抗体可在体外抑制血管生成。本研究旨在确定通过重组技术产生的腹腔疾病患者来源的靶向TG2的单克隆抗体是否具有与血清来源的腹腔疾病自身抗体相似的抗血管生成作用。此外，我们研究了单克隆患者自身抗体是否调节内皮细胞TG2活性，以及这种调节是否与抗血管生成作用相关。
使用三维血管生成细胞培养模型研究腹腔疾病患者来源的靶向TG2的单克隆抗体对内皮细胞小管形成的影响。通过活细胞酶联免疫吸附测定法测定内皮细胞TG2酶活性。
通过重组技术产生的腹腔疾病患者来源的靶向TG2的单克隆抗体抑制内皮细胞小管形成并增强TG2的交联活性。当在自身抗体存在下使用定点不可逆TG2抑制剂抑制这种酶活性时，体外血管生成恢复到对照水平。
由于我们发现内皮细胞血管生成与TG2活性之间存在显著负相关，我们认为腹腔疾病患者来源的靶向TG2的自身抗体的抗血管生成作用是由TG2增强的酶活性发挥的。
