Anti-SSA/Ro and anti-SSB/La autoantibodies bind the surface of apoptotic fetal cardiocytes and promote secretion of TNF-alpha by macrophages.
Despite the near universal association of congenital heart block and maternal Abs to SSA/Ro and SSB/La, the intracellular location of these Ags has made it difficult to substantiate their involvement in pathogenicity. To define whether components of the SSA/Ro-SSB/La complex, which translocate during apoptosis, are indeed accessible to extracellular Abs, two approaches were taken: immunoprecipitation of surface biotinylated proteins and scanning electron microscopy. Human fetal cardiocytes from 16-24-wk abortuses were cultured and incubated with staurosporine to induce apoptosis. Surface biotinylated 48-kDa SSB/La was reproducibly immunoprecipitated from apoptotic, but not nonapoptotic cardiocytes. Surface expression of SSA/Ro and SSB/La was further substantiated by scanning electron microscopy. Gold particles (following incubation with gold-labeled sera containing various specificities of anti-SSA/Ro-SSB/La Abs and murine mAb to SSB/La and 60-kDa SSA/Ro) were consistently observed on early and late apoptotic cardiocytes. No particles were seen after incubation with control antisera. To evaluate whether opsonized apoptotic cardiocytes promote inflammation, cells were cocultured with macrophages. Compared with nonapoptotic cardiocytes or apoptotic cardiocytes incubated with normal sera, apoptotic cardiocytes preincubated with affinity-purified Abs to SSB/La, 52-kDa SSA/Ro, or 60-kDa SSA/Ro increased the secretion of TNF-alpha from cocultured macrophages. In summary, apoptosis results in surface accessibility of all SSA/Ro-SSB/La Ags for recognition by circulating maternal Abs. It is speculated that in vivo such opsonized apoptotic cardiocytes promote an inflammatory response by resident macrophages with damage to surrounding conducting tissue.
Miranda-Carús
M E
ME
Department of Rheumatology, Hospital for Joint Diseases, New York University School of Medicine, New York, NY 10003, USA.
Askanase
A D
AD
Clancy
R M
RM
Di Donato
F
F
Chou
T M
TM
Libera
M R
MR
Chan
E K
EK
Buyon
J P
JP
D000974:Antibodies, Antinuclear / Q000378:metabolism*; D017209:Apoptosis / Q000276:immunology*; D001666:Binding Sites, Antibody; D019921:Biotinylation; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D011232:Chemical Precipitation; D018920:Coculture Techniques; D005260:Female; D005318:Fetal Heart / Q000166:cytology / Q000276:immunology* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D008264:Macrophages / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D009206:Myocardium / Q000166:cytology / Q000276:immunology* / Q000378:metabolism / Q000648:ultrastructure; D011247:Pregnancy; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D000974:Antibodies, Antinuclear; C035356:SS-A antibodies; C035357:SS-B antibodies; D014409:Tumor Necrosis Factor-alpha