Human immunoglobulin G autoantibodies to the thyrotropin receptor from Epstein-Barr virus-transformed B lymphocytes: Characterization by immunoprecipitation with recombinant antigen and biological activity

被引:29
作者
Morgenthaler, NG
Kim, MR
Tremble, J
Huang, GC
Richter, W
Gupta, M
Scherbaum, WA
McGregor, AM
Banga, JP
机构
[1] UNIV LONDON KINGS COLL, SCH MED, DEPT MED, LONDON SE5 9PJ, ENGLAND
[2] CLEVELAND CLIN FDN, ENDOCRINE IMMUNOL LAB, CLEVELAND, OH USA
[3] UNIV LEIPZIG, DEPT INTERNAL MED 3, D-7010 LEIPZIG, GERMANY
[4] UNIV ULM, D-7900 ULM, GERMANY
关键词
D O I
10.1210/jc.81.9.3155
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The TSH receptor (TSH-R) is the target antigen for disease-related autoantibodies in Graves' disease and primary myxoedema, but the repertoire of the antibodies or the nature of the precise antigenic epitopes is not known. We have immortalized peripheral blood B cells from six different autoimmune thyroid disease patients with Epstein-Barr virus and selected IgG-producing B cells by magnetic selection on anti-IgG-coated beads. Purified recombinant insect cell-derived extracellular region of TSH-R was used to identify the positive wells for expansion in culture. Stable B cell lines (n = 11) were obtained, which after limiting dilution led to two stable B cell clones. B cell lines and clones secreted IgG antibody that were shown to react biochemically with metabolically labeled or in vitro translated, nascent extracellular region of TSH-R, giving strong confirmatory evidence of the presence of anti-TSH-R antibody. Supernatants from lines contained thyroid-stimulating activity, thyroid-blocking activity (as assessed by inhibition of TSH-mediated cAMP stimulation), or both of these activities. Interestingly, antibodies with stimulating activity were generated from a primary myxoedema patient, and antibodies of blocking specificities were obtained from newly diagnosed thyrotoxic Graves' disease patients. Our results favor a fine balance between stimulating and blocking autoantibody activities in determining the clinical presentation observed in patients with autoimmune thyroid disease patients who have these antibodies present in their serum.
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页码:3155 / 3161
页数:7
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