Celiac disease and pregnancy outcome: new evidences of anti-transglutaminase antibodies effect on human endometrial endothelial angiogenesis

Celiac disease (CD) is an autoimmune enteropathy triggered in susceptible individuals by gluten. CD patients have circulating anti-transglutaminase antibodies (anti-TGA), mediators of intestinal and extra-intestinal manifestations of CD. The epidemiological association between maternal CD and increased frequency of obstetric failure is well established, although the pathogenic mechanisms are still poorly understood. Recently, we observed that anti-TGA IgG bind to trophoblast cell cultures causing an apoptotic damage and reducing cytotrophoblast invasiveness. Since the placentation process requires a proper endometrial angiogenesis, our aim was to study the possible effect of anti-TGA on angiogenesis process.Materials and Methods IgG and IgA anti-TGA were purified from serum of CD patient on a gluten-containing diet through commercial kit. Endometrial tissues were collected from fertile women undergoing hysterectomy for fibroid uterus. HEEC were isolated and purified through tissue digestion, centrifugation, immune selection, and characterized by cytometry. In vitro binding assay on HEEC culture of IgG and IgA monoclonal and polyclonal anti-TGA was performed through ELISA and immunofluorescence staining. HEEC capability to form capillary-like tube structures in vitro was evaluated. In vivo angiogenesis in subcutaneous angioreactors treated with TGA and implanted in mice was analyzed. Phalloidin-FITC staining of HEEC cytoskeleton F-actin fibres was performed and studied by inverted confocal microscope.Results We showed that anti-TGA: bind to HEEC in a dose and time dependent manner; significantly decrease the number and the total length of the tubules formed by HEEC; determine a disorganization of cytoskeleton F-actin fibres in HEEC [Figure 1]; when inoculated in mice, cause a significant reduction of newly formed blood vessels. Conclusions We firstly demonstrated an anti-TGA-mediated angiogenesis impairment, in term of HEEC in vitro angiogenesis reduction through cytoskeleton disorganization, and in vivo angiogenetic capability decrease in mice. Our results provide an additional pathogenic mechanism of placental damage in CD able to explain its epidemiological association with adverse pregnancy outcomes.