Congenital Heart Block Maternal Sera Autoantibodies Target an Extracellular Epitope on the [alpha].sub.1G T-Type Calcium Channel in Human Fetal Hearts

• Published in: PloS one 2013-09, Vol.8 (9), p.e72668
• Main Authors: Strandberg, Linn S, Cui, Xuezhi, Rath, Arianna, Liu, Jie, Silverman, Earl D, Liu, Xiaoru, Siragam, Vinayakumar, Ackerley, Cameron, Su, Brenda Bin, Yan, Jane Yuqing, Capecchi, Marco, Biavati, Luca, Accorroni, Alice, Yuen, William, Quattrone, Filippo, Lung, Kalvin, Jaeggi, Edgar T, Backx, Peter H, Deber, Charles M, Hamilton, Robert M
• Format: Article
• Language: English
• Subjects: Antigenic determinants; Autoantibodies; Autoimmunity; Calcium channels; Comparative analysis; Enzyme-linked immunosorbent assay; Genetic disorders; Heart block; Peptides; Pregnancy
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0072668

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 [alpha].sub.1G may be a fetal target of maternal sera autoantibodies in CHB. We demonstrate differential mRNA expression of the T-type calcium channel CACNA1G ([alpha].sub.1G 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 [alpha].sub.1G epitope on the surfaces of cardiomyocytes as well as reactivity of maternal serum from CHB affected pregnancies to the [alpha].sub.1G protein. By ELISA we demonstrated maternal sera reactivity to [alpha].sub.1G 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 [alpha].sub.1G repeat I). Maternal sera from CHB affected pregnancies also reacted more weakly to the homologous region (7/15 amino acids conserved) of the [alpha].sub.1H channel. Electrophysiology experiments with single-cell patch-clamp also demonstrated effects of CHB maternal sera on T-type current in mouse sinoatrial node (SAN) cells. Taken together, these results indicate that CHB maternal sera antibodies readily target an extracellular epitope of [alpha].sub.1G T-type calcium channels in human fetal cardiomyocytes. CHB maternal sera also show reactivity for [alpha].sub.1H suggesting that autoantibodies can target multiple fetal targets.