title: Congenital heart block maternal sera autoantibodies target an extracellular epitope on the α1G T-type calcium channel in human fetal hearts.
abstract: 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 α1G may be a fetal target of maternal sera autoantibodies in CHB.
  
  
  RESULTS
  We demonstrate differential mRNA expression of the T-type calcium channel CACNA1G (α1G 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 α1G epitope on the surfaces of cardiomyocytes as well as reactivity of maternal serum from CHB affected pregnancies to the α1G protein. By ELISA we demonstrated maternal sera reactivity to α1G 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 α1G repeat I). Maternal sera from CHB affected pregnancies also reacted more weakly to the homologous region (7/15 amino acids conserved) of the α1H 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.
  
  
  CONCLUSIONS
  Taken together, these results indicate that CHB maternal sera antibodies readily target an extracellular epitope of α1G T-type calcium channels in human fetal cardiomyocytes. CHB maternal sera also show reactivity for α1H suggesting that autoantibodies can target multiple fetal targets.
authors.affiliation: Department of Physiology and Experimental Medicine, The Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: Linn S
authors.initials: LS
authors.lastname: Strandberg
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Cui
authors.forename: Arianna
authors.initials: A
authors.lastname: Rath
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.forename: Earl D
authors.initials: ED
authors.lastname: Silverman
authors.forename: Xiaoru
authors.initials: X
authors.lastname: Liu
authors.forename: Vinayakumar
authors.initials: V
authors.lastname: Siragam
authors.forename: Cameron
authors.initials: C
authors.lastname: Ackerley
authors.forename: Brenda Bin
authors.initials: BB
authors.lastname: Su
authors.forename: Jane Yuqing
authors.initials: JY
authors.lastname: Yan
authors.forename: Marco
authors.initials: M
authors.lastname: Capecchi
authors.forename: Luca
authors.initials: L
authors.lastname: Biavati
authors.forename: Alice
authors.initials: A
authors.lastname: Accorroni
authors.forename: William
authors.initials: W
authors.lastname: Yuen
authors.forename: Filippo
authors.initials: F
authors.lastname: Quattrone
authors.forename: Kalvin
authors.initials: K
authors.lastname: Lung
authors.forename: Edgar T
authors.initials: ET
authors.lastname: Jaeggi
authors.forename: Peter H
authors.initials: PH
authors.lastname: Backx
authors.forename: Charles M
authors.initials: CM
authors.lastname: Deber
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hamilton
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001283:Atrioventricular Node / Q000187:drug effects / Q000378:metabolism; D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020747:Calcium Channels, T-Type / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D005110:Extracellular Space; D005260:Female; D005318:Fetal Heart / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression; D006327:Heart Block / Q000151:congenital* / Q000235:genetics / Q000276:immunology; D006801:Humans; D008297:Male; D008431:Maternal-Fetal Exchange / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D032383:Myocytes, Cardiac / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000276:immunology; D011247:Pregnancy; D011817:Rabbits
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; C121855:CACNA1G protein, human; D002121:Calcium Channel Blockers; D020747:Calcium Channels, T-Type; D000939:Epitopes; D010455:Peptides
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