The effects of nonionic polymers on thyroid stimulation and TSH receptor binding by the human monoclonal TSH receptor autoantibody M22.
BACKGROUND
Nonionic polymers such as polyethylene glycol (PEG), polyvinyl alcohol (PVA), and dextran amplify the ability of thyroid stimulating antibodies (TSAbs) from patients with Graves' disease to stimulate cAMP production in thyroid cells. Therefore we sought to determine if nonionic polymers also augment the effects of the human thyroid stimulating monoclonal antibody (M22) on isolated thyroid cells.


METHODS
The ability of nonionic polymers to alter the effects of M22 on certain parameters in porcine thyroid cells was examined. These parameters were augmentation of cAMP production (TSAb activity), inhibition of bovine thyrotropin (bTSH)-induced cAMP production (TBAb activity), and inhibition of bTSH binding to the TSH receptor (TSHR) (TBI activity).


RESULTS
Stimulation of cAMP production by M22 in porcine thyroid cells was augmented by PEG, PVA, and dextran in a manner similar to that of Graves' serum. In contrast, TSH-stimulated cAMP production was not increased by nonionic polymers. M22-stimulated cAMP production was completely inhibited by the sera of patients with TBAb activity, and this inhibition was diminished by PEG. M22 and TBI activity in first and second generation assays and this activity was not affected by PEG. Binding of biotin-M22 to TSHR-coated plate wells (third generation assay) was not significantly increased by co-incubation with polymers. PEG augmented the binding of (125)I-M22 to TSHR-coated tubes by twofold, but this was associated with a threefold increase in nonspecific binding. There was no increase in total and nonspecific (125)I-TSH binding. This means that PEG has less than a twofold augmentative effect on (125)I-M22 binding to the TSHR.


CONCLUSIONS
Nonionic polymers have similar effects in augmenting cAMP production in porcine thyroid cells in response to stimulation either by M22 or Graves' disease serum. The mechanism of this effect on the thyroid stimulating activity of M22 is unclear. The hypothesis that nonionic polymers augment M22 thyroid stimulation by increasing the mass of M22-occupied TSH receptors is not supported by the present study.
Ochi
Yukio
Y
Research Institute for Production Development, Kyoto, Japan. k-y-ochi@blue.plala.or.jp
Hamazu
Masanari
M
Kajita
Yoshihiro
Y
Hachiya
Takashi
T
Miyata
Takeshi
T
Barrett
Carol
C
Smith
Bernard Rees
BR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D001798:Blood Proteins / Q000494:pharmacology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D003911:Dextrans / Q000494:pharmacology*; D006111:Graves Disease / Q000097:blood; D006801:Humans; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011142:Polyvinyl Alcohol / Q000494:pharmacology*; D011485:Protein Binding; D011989:Receptors, Thyrotropin / Q000276:immunology / Q000378:metabolism*; D013552:Swine; D013961:Thyroid Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013972:Thyrotropin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D003911:Dextrans; D011989:Receptors, Thyrotropin; D011092:Polyethylene Glycols; D013972:Thyrotropin; D011142:Polyvinyl Alcohol; D000242:Cyclic AMP