Hydrogen peroxide-induced production of a 40 kDa immunoreactive thyroglobulin fragment in human thyroid cells: the onset of thyroid autoimmunity?
We recently reported that, during in vitro thyroid-hormone synthesis, H(2)O(2) stress cleaved thyroglobulin (Tg) into C-terminal peptides. These peptides were found to contain the immunodominant region of Tg recognized by Tg autoantibodies from patients with an autoimmune thyroid disease. To test the hypothesis that Tg fragmentation is an early upstream initiating event involved in Tg autoimmune response and the consequence of oxidative injuries, we studied the effect of H(2)O(2) stress on human thyroid cells. In culture conditions allowing Tg synthesis and iodine organification by the cells, we found that bolus addition of increasing millimolar doses of H(2)O(2) induced a dose-response appearance of floating cells in the culture medium. These cells apparently resulted from a necrotic process, and they bore iodinated Tg fragments. These fragments were found to be similar to those previously obtained in vitro from purified Tg. In both cases, Tg peptides were recognized by a well-defined monoclonal antibody directed to the immunodominant region of Tg. The smallest immunoreactive Tg peptide had a molecular mass of 40 kDa and entered human thyrocytes more efficiently than the entire Tg. These data suggest that thyrocytes exposed to locally increased H(2)O(2) doses accumulate fragmented Tg for further delivery into surrounding living thyrocytes in the course of an autoimmune response.
Duthoit
C
C
U555 INSERM, Faculté de Médecine Timone, Université de la Méditerranée, Marseille, France.
Estienne
V
V
Giraud
A
A
Durand-Gorde
J M
JM
Rasmussen
A K
AK
Feldt-Rasmussen
U
U
Carayon
P
P
Ruf
J
J
D000236:Adenoma / Q000601:surgery; D000328:Adult; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D018828:Immunoglobulins, Thyroid-Stimulating / Q000276:immunology; D008875:Middle Aged; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism*; D011485:Protein Binding; D013954:Thyroglobulin / Q000378:metabolism*; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism*; D013964:Thyroid Neoplasms / Q000601:surgery; D013967:Thyroiditis, Autoimmune / Q000276:immunology*
D018828:Immunoglobulins, Thyroid-Stimulating; D010446:Peptide Fragments; D013954:Thyroglobulin; D006861:Hydrogen Peroxide