Antigenic binding sites of anti-protein S autoantibodies in patients with recurrent pregnancy loss

Protein S (PS) deficiency is a risk factor for adverse pregnancy outcomes including recurrent pregnancy loss. Several studies have shown that the presence of anti-PS autoantibodies (anti-PS) leads to an acquired PS deficiency. Hence, an epitope mapping study was conducted to know the pathogenesis of anti-PS in patients with recurrent pregnancy loss.

These results suggest that anti-PS in patients with recurrent pregnancy loss recognize EGF-like domains in PS. Interestingly, anti-PS also recognized EGF family proteins. Anti-PS in patients with recurrent pregnancy loss may be associated with not only thrombophilia but also the disruption of the EGF system.

PS was treated with thrombin to divide the protein into γ-carboxyglutamic acid (Gla) domain and Gla-domain free PS. For the preparation of fragments of epidermal growth factor (EGF)-like domains (EGF1-4), PS was subjected to proteolysis using lysyl endopeptidase. The epitopes were identified in immunoblot. Whether anti-PS recognized EGF family proteins in anti-PS-positive patients was also examined.

Anti-PS recognized Gla-domain free PS, especially the three fragments of EGF-like domains, EGF1-2, EGF3-4, and EGF1-4. Anti-PS recognized recombinant human EGF. Anti-PS and polyclonal antibodies to recombinant human EGF recognized PS in the absence of Ca2+ but not in the presence of Ca2+. In competitive inhibition studies, polyclonal antibodies to recombinant mouse EGF blocked anti-PS binding to PS in a concentration-dependent manner. Conclusions: These results suggest that anti-PS in patients with recurrent pregnancy loss recognize EGF-like domains in PS. Interestingly, anti-PS also recognized EGF family proteins. Anti-PS in patients with recurrent pregnancy loss may be associated with not only thrombophilia but also the disruption of the EGF system.