Contributions
Abstract: P1904
Type: Poster
Abstract Category: Late breaking news
Background: Antiphospholipid antibodies (aPL) are known to occur in Multiple Sclerosis (MS) patients, even though their exact role remains unclear. aPL are key in the formation of thrombi in the Antiphospholipid Syndrome (APS) with most studies now focusing on the pro-thrombotic role of antibodies targeting Domain I of β2-Glycoprotein I (anti-DI). In a previous study we have shown the elevated levels of anti-DI and anti-Cardiolipin (anti-CL) antibodies in MS patients compared to Healthy Controls (HC). In the present study, we investigate the possible pro-inflammatory effects of IgG antibodies from MS patients compared to HC in vitro.
Methods: Total IgG was purified from ten MS patients that were positive for anti-DI and nine MS patients that were positive for anti-CL antibodies; along with age and gender matched HC groups (negative for both aPL). IgG samples were pooled together in groups according to positivity for either aPL and tested for endotoxin. 100 µg/ml of IgG was used to stimulate an astrocytic cell line, U87 at different time points from 10 minutes up to 12 hours. Subsequently, the cells were lysed and cell lysates were quantified and analyzed by immunoblot using antibodies for p38 MAPK and p65 NF-κB.
Results: Phosphorylation of p38 MAPK and p65 NF-κB increased two-fold with IgG from MS patients compared to HC after 1 hour (for p65 NF-κB, p< 0.05). This was also observed using IgG from MS patients positive for IgG anti-DI. Overall, there was a considerable difference in phosphorylation of p65 NF-κB, and a significantly higher phosphorylation of p38 MAPK in astrocytes treated with IgG from MS patients (positive for either aPL) compared to both untreated cells, or cells treated with IgG from HC
(p< 0.05 for both).
Conclusion: IgG from MS patients positive for aPL, activate inflammatory signaling and may signify a role in pro-thrombotic conditions in MS.
Disclosure: All authors have nothing to disclose.
Abstract: P1904
Type: Poster
Abstract Category: Late breaking news
Background: Antiphospholipid antibodies (aPL) are known to occur in Multiple Sclerosis (MS) patients, even though their exact role remains unclear. aPL are key in the formation of thrombi in the Antiphospholipid Syndrome (APS) with most studies now focusing on the pro-thrombotic role of antibodies targeting Domain I of β2-Glycoprotein I (anti-DI). In a previous study we have shown the elevated levels of anti-DI and anti-Cardiolipin (anti-CL) antibodies in MS patients compared to Healthy Controls (HC). In the present study, we investigate the possible pro-inflammatory effects of IgG antibodies from MS patients compared to HC in vitro.
Methods: Total IgG was purified from ten MS patients that were positive for anti-DI and nine MS patients that were positive for anti-CL antibodies; along with age and gender matched HC groups (negative for both aPL). IgG samples were pooled together in groups according to positivity for either aPL and tested for endotoxin. 100 µg/ml of IgG was used to stimulate an astrocytic cell line, U87 at different time points from 10 minutes up to 12 hours. Subsequently, the cells were lysed and cell lysates were quantified and analyzed by immunoblot using antibodies for p38 MAPK and p65 NF-κB.
Results: Phosphorylation of p38 MAPK and p65 NF-κB increased two-fold with IgG from MS patients compared to HC after 1 hour (for p65 NF-κB, p< 0.05). This was also observed using IgG from MS patients positive for IgG anti-DI. Overall, there was a considerable difference in phosphorylation of p65 NF-κB, and a significantly higher phosphorylation of p38 MAPK in astrocytes treated with IgG from MS patients (positive for either aPL) compared to both untreated cells, or cells treated with IgG from HC
(p< 0.05 for both).
Conclusion: IgG from MS patients positive for aPL, activate inflammatory signaling and may signify a role in pro-thrombotic conditions in MS.
Disclosure: All authors have nothing to disclose.