Congenital Heart Block Maternal Sera Autoantibodies Target an Extracellular Epitope on the a1G T-Type Calcium Channel in Human Fetal Hearts

Background: Congenital heart block (CHB) is a transplacentally acquired autoimmune disease associated with anti-Ro/SSA and anti-La/SSB maternal autoantibodies and is characterized primarily by atrioventricular (AV) block of the fetal heart. This study aims to investigate whether the T-type calcium channel subunit a1G may be a fetal target of maternal sera autoantibodies in CHB. Methodology/Principal Findings: We demonstrate differential mRNA expression of the T-type calcium channel CACNA1G (a1G gene) in the AV junction of human fetal hearts compared to the apex (18–22.6 weeks gestation). Using human fetal hearts (20–22 wks gestation), our immunoprecipitation (IP), Western blot analysis and immunofluorescence (IF) staining results, taken together, demonstrate accessibility of the a1G epitope on the surfaces of cardiomyocytes as well as reactivity of maternal serum from CHB affected pregnancies to the a1G protein. By ELISA we demonstrated maternal sera reactivity to a1G was significantly higher in CHB maternal sera compared to controls, and reactivity was epitope mapped to a peptide designated as p305 (corresponding to aa305–319 of the extracellular loop linking transmembrane segments S5–S6 in a1G repeat I). Maternal sera from CHB affected pregnancies also reacted more weakly to the homologous region (7/15 amino acids conserved) of the a1H channel. Electrophysiology experiments with single-cell patch-clamp also demonstrated effect