Inhibitory effect of thyroid blocking antibody (TBAb) on the thyroid stimulatory effect of human chorionic gonadotropin (HCG) and equine chorionic gonadotropin (ECG).

We examined the inhibitory effect of thyroid blocking antibody (TBAb) on the thyroid stimulating activity of human chorionic gonadotropin (HCG) and equine CG (ECG). Five TBAb positive sera obtained from patients who had been hypothyroid but were currently on T4 treatment. The TSH binding inhibitory immunoglobulin (TBII) activities of the sera were 60-160 IU/L. Inhibition of TSH binding to the TSH receptor (TSHR) [TSH binding inhibition (TBI) activity] of HCG or ECG, and inhibition of TBAb on HCG or ECG-stimulated cAMP production were examined. Both HCG and ECG preparations showed weak TBI activity in the presence of small amounts of protein [bovine serum albumin (BSA)] but were negative in the presence of large amounts of protein [normal human serum (NHS) or BSA]. Four thousand IU/mL of HCG and ECG preparation caused cAMP production similar to 100 microU/mL of bovine (b) TSH. The inhibitory effect of TBAb on cAMP production by this amount of HCG or ECG was then examined. The inhibitory effect of TBAb on cAMP production by HCG and ECG was similar to bTSH, and TBAb positive sera with more than 40 IU/L TBII activity completely blocked cAMP production by HCG, ECG and bTSH. This suggests that common alpha -subunit of both HCG and TSH are involved in the inhibitory effect of TBAb. Previous reports demonstrated that the thyroid stimulating activity of thyroid stimulating antibody (TSAb) was blocked by deglycosylated HCG (competitive antagonist of TSH binding to TSHR). The fact and our present study suggest that TSH, HCG ECG, TSAb and TBAb have a similar binding site (alpha-subunit-mimicking binding site) on the TSH receptor.

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. CONCLUSION: 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.

Increased receptor binding by bovine (b) TSH bound to monoclonal antibody to bTSHbeta-subunit.

TSH receptor (R) binding and cAMP production by bovine (b) TSH-bound to a monoclonal antibody (MoAb) or polyclonal antibody (Ab) to bTSH were examined, using TSH receptor (R) coating tube and porcine thyroid cells. (125) I-bTSH bound-to MoAbs to bTSH(alpha) or discontinuous type MoAb showed TSHR binding (10%) similar to intact (125) I-bTSH. TSHR binding was completely decreased (<2%) when (125) I-bTSH was bound by polyclonal Abs to bTSH(alpha) in Graves' patient or rabbit polyclonal Abs to bTSH. When either of the two MoAb (No. 1 and 2) to bTSH(beta) was bound to (125) I-bTSH, TSHR binding was 4 times higher (40%) compared to intact (125) I-bTSH. Binding of another MoAb (No. 3) caused no increased binding. TSHR binding of intact (125) I-bTSH was decreased from 10% to 2% by excess amounts of bTSH. Binding of (125) I-bTSH bound to MoAb to bTSH(beta) (No. 1 and 2) decreased from 40% to 30% by excess amounts of bTSH. When (125) I-bTSH bound-Fab of MoAb was used, the binding was reduced from 30 to 10% (No. 1) and from 25 to 6% (No. 2), respectively. In contrast, cAMP production by bTSH was decreased by pre-binding of all MoAbs and polyclonal Abs. Binding of (125) I-MoAb to bTSH (beta) to a synthetic peptide array of bTSH (beta) sequence was examined by the radioautography. The epitope of MoAb to bTSH(beta) was suggested to be LPK (beta 42-44) for No. 1, KLF (beta 39-41) for No. 2 and PKYA (beta 43-46) for No. 3, respectively, although the existence of discontinuous epitope could not be ruled out. The increased TSHR binding and the decreased cAMP production by bTSH bound to MoAbs may be due to the conformational change of TSH molecule or TSHR by binding of both bTSH and MoAb.

[Examination of attenuation correction using low-dose computed tomography].

Attenuation correction is necessary for the reconstruction of SPECT images. One report has mentioned that attenuation correction by X-ray computed tomography (CT) is effective for a non-uniform attenuation body. We examined the effect of attenuation correction on SPECT images by changing the scanning conditions of CT, and evaluated the possibility of attenuation correction by low-dose CT. The phantom was scanned under several X-ray tube conditions varying from 80 kV to 135 kV and from 7.5 mAs to 200 mAs. We obtained equations of attenuation correction based on the Hounsfield Unit (HU) units of each pixel and compared the effects of attenuation correction. The results showed that the equation for attenuation correction under each condition did not vary significantly, and the effects of attenuation correction by the equations did not vary significantly between CT of low dose and that of clinical dose. This result suggest that the attenuation correction obtained by low-dose CT was equal to that obtained by the clinical dose. In conclusion, it seemed that the equation and map of attenuation correction matched with each radionuclide yielded more adequate attenuation correction than conventional methods.

Antibody to immunoglobulin G and polyethylene glycol augment cyclic adenosine monophosphate production by TSH receptor antibody bound to porcine thyroid cells.

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 degrees 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 degrees 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.

[Introduction of manual for the management radioactive medical waste].

Societies concerned with radioactive rays and nuclear medicine have recently highlighted the necessity of managing radioactive medical waste resulting from nuclear medicine examinations. We introduce a manual that we have created and explain its use in decision-making and management practices aimed at the reduction of radioactive medical waste at hospitals that have not yet solved this problem. We hope that our manual will help in reducing this medical waste.

Demonstration of anti-TSH antibody in TSH binding inhibitory immunoglobulin-positive sera of patients with Graves' disease.

OBJECTIVES: The presence of anti-TSH antibodies in Graves' patients with unusually low TSH binding inhibitory immunoglobulin (TBII) has been reported. Recently, we found the first case of an anti-TSH antibody in TBII-positive sera of patients with Graves' disease. The prevalence and immunological specificity of this anti-TSH antibody were examined. DESIGN: The presence of 125I-bovine(b) TSH binding antibody in TBII positive serum was examined by prolonged incubation of more than 1 day because only weak binding occurred after 1 h incubation at 37 degrees C. The clinical course of these patients and binding characteristics of anti-bTSH antibody were examined. RESULTS: The corrected method-TBII activity (%)[1 - (a - b)/(c - d)] x 100 and the standard method-TBII activity (%) [1 - (a - d)/(c - d)] x 100 [a, 125I-bTSH binding with TSH receptor (R) in the presence of test serum; b, 125I-bTSH binding with test serum; c, 125I-bTSH binding with TSH R in the presence of normal serum; d, 125I-bTSH binding with normal serum] were calculated. The corrected method-TBII activity was always higher than the standard method-TBII activity in anti-bTSH antibody-positive serum. Anti-bTSH antibody-positive cases in TBII-positive Graves' disease were found in approximately 1% of Graves' patients. Anti-bTSH antibodies were confirmed as IgG from the increase of precipitated radioactivity by adding rabbit antihuman IgG antibody after the incubation of 125I-bTSH with test serum. These antibodies bind with not only bTSH, bTSH(alpha) and bLH, but also porcine (p)TSH, pTSH(alpha) and pFSH. However, these antibodies did not bind with human TSH. Binding of 125I-bTSH with patient's serum was neither inhibited by other Graves' thyroid stimulating antibody (TSAb), nor thyroid blocking antibody (TBAb) in primary hypothyroidism. CONCLUSIONS: The presence of anti-bTSH antibody in TBII-positive serum of high titre means that TBII-positive sera cannot rule out the absence of anti-bTSH. Thus, determination of 125I-bTSH binding with test serum in TSH receptor assays is necessary to determine the precise TBII activity and to detect anti-bTSH antibody.