Antibody to Immunoglobulin G and Polyethylene Glycol Augment Cyclic Adenosine Monophosphate Production by TSH Receptor Antibody Bound to Porcine Thyroid Cells

Abstract

Anti-immunoglobulin G (IgG) augments cyclic adenosine monophosphate (cAMP) production by thyroid-blocking antibody (TBAb) bound to porcine thyroid cells (PTC). This is described as a conversion phenomenon. We reported the effect of polyethylene glycol (PEG) to augment thyroid-stimulating antibody (TSAb) activity in a PTC assay. In the present experiment we examined the effect of anti-immunoglobulin G (IgG) and PEG on cAMP production from TBAb or TSAb bound to PTC. TBAb bound to PTC was separated from unbound TBAb by centrifugation after a first incubation (0.5 hour at 37°C) of TBAb-IgG with PTC. TBAb bound to PTC were incubated with anti-human (h)IgG or hIgG fragments [F(ab′)₂, Fc, Fd, H chain or L-chain] for 4 hours at 37°C in the second incubation. Anti-IgG or anti-IgG fragments increased cAMP production. No conversion was caused by protein A, protein L, or PEG (5%). PEG did not augment cAMP production by these IgG antibodies. PEG augmented cAMP production during incubation of TSAb-IgG bound to PTC, but anti-IgG did not. PEG significantly augmented cAMP production by coincubation of TSAb-IgG bound to PTC and the unbound TSAb-IgG (obtained from the first incubation). A similar augmentative effect of PEG was also observed in experiments using TSAb-F(ab′)₂ and TSAb-Fab. cAMP production by PTC bound by both TBAb- and TSAb-IgG was increased by co-incubation with anti-IgG, but was not increased by PEG. In conclusion, anti-IgG specifically increased cAMP production from TBAb bound to PTC (conversion phenomenon) and PEG specifically increased cAMP production by TSAb bound to PTC. Different mechanisms enhance cAMP production by TSAb and conversion of TBAb.