BACKGROUND: The Chinese hamster ovary cells transfected with human TSH receptor cDNA (hTSHR-CHO), expressing functional human TSH receptors, are known to be more sensitive in detection of thyroid stimulating antibodies than FRTL-5 cells. There has been no report on the usefulness of these cells to measure thyroid stimulation blocking antibody (TSBAb) activity which is frequently found in patients with primary myxedema, METHODS: We established the optimal assay condition of measurement of TSBAb using hTSHR-CHO cells, and simultaneously measured TSBAb activities with FRTL-5 cells and with hTSHR-CHO cells in 49 patients with primary myxedema, compared them with their thyrotropin binding inhibitor immunoglobulin (TBII) activities. RESULTS: 1) hTSHR-CHO cells specifically bound bTSH and were stimulated by bTSH in terms of cyclic AMP generation in a dose dependent manner. 2) Myxedema IgG suppressed TSH-stimulated cAMP production of hTSHR-CHO cells in a dose dependent manner reaching plateau at the concentration of I g/L. Normal pooled IgG has no suppressive action at the concentration of less than 1 g/L, but caused significant suppression at the concentration of greater than 1g/L. 3) TSBAb activities measured by hTSHR-CHO cells in 49 patients with primary myxedema were as follows: Four of 25 TBII-negative cases (16%) and 22 of 24 TBII-positive cases (92%) had TSBAb activities. Most of TSBAb positive patients (95%), especially in TBII positive cases, showed very high activities of more than 90%. 4) TSBAb activities measured by hTSHR-CHO cells and those by FRTL-5 cells were both positive in 24 patients (49%), both negative in 18 patients (37%), and were discrepant in 7 patients (14%). The TSBAb activities measured with hTSHR-CHO cells and those measured with FRTL-5 cells were significantly correlated (r=0.71, p< 0.01). 5) Forty five percent of patients with primary myxedema had all of 3 kinds of activities (TBII, hTSHR-CHO cell TSBAb, FRTL-5 cell TSBAb), 37% of them had none of 3 activities and 18% of them had 1 or 2 kinds of activities only. CONCLUSION: The usefulness of hTSHR-CHO cells in measurements of TSBAb activities were confirmed. The TSBAb activities of most patients with primary myxedema measured by hTSHR-CHO cells were concordant with those measured by FRTL-5 cells. However, a small subset of patients (18%) had discrepant results in assays of TSH receptor antibodies according to the differences in TSH receptors (rat, human and porcine) used in assay. Such discrepancy may be explained by heterogeneity in epitopes for blocking TSH receptor antibodies.
Animals ; Antibodies ; Asian Continental Ancestry Group* ; Cricetinae ; Cricetulus* ; Cyclic AMP ; DNA, Complementary ; Epitopes ; Female ; Humans ; Humans* ; Hypothyroidism* ; Immunoglobulin G ; Immunoglobulins ; Immunoglobulins, Thyroid-Stimulating ; Myxedema ; Ovary* ; Population Characteristics ; Receptors, Thyrotropin* ; Thyroid Gland* ; Thyrotropin

No abstract available.
Capsid Proteins* ; Capsid* ; Genome*

BACKGROUND: TSH receptor blocking antibody (TRBAb) is a pathogenic factor in the vast majority of patients with primary myxedema. It has been reported that TRBAbs are found in some patients with chronic goitrous autoimmune thyroiditis (Hashimoto's thyroiditis), but the significance or the role of TRBAb in Hashimotos thyroiditis is not clear, We recently reported that hTSHR-CHO cells which express the functional human TSH receptors are more sensitive and are better in detecting functional TSH receptor antibodies in Graves patients than FRTL-5 cells. We are to investigate the biological role of TRBAb in Hashimotos thyroiditis by measuring thyroid stimulation blocking antibody (TSBAb) activities of Hashimoto's IgG's using hTSHR-CHO cells. Moreover, we are to see if there is any difference in epitope recognition between Hashimotos TRBAb and myxedema's TRBAb by measuring TSBAb activities with mutant receptor expressing cell lines, Mcl+2 and Mc 2 in those patients. METHOD: We measured TSBAb activities of IgGs from patients with primary myxedema (PM, n= 10) and those with hypothyroid (n 20) or euthyroid (n 17) Hashimoto's thyroiditis (HT) using wild type hTSHR-CHO cells (WT) and two chimeric receptor expressing cell lines, Mcl+2 and Mc2. RESULTS: TSBAb activities measured by WT were higher in hypothyroid HT than in euthyroid HT (30.0+-23.2% vs. 6.1+-28.7, p<0.05), and TSBAb-positive rate tend to be higher in the former (20%, 5/20) than in the latter (0%, 0/17, p=0.07). TRBAbs from PM (n=4) had high TBII activities and had persistent blocking activities despite of the replacement of amino acid residue 8~165 of extracellular domain of TSHR with those of rat LH/CGR (Mcl +2). However, TRBAbs from HT (n=4) had no TBII activity at all and lost blocking activities when measured with Mcl+2. CONCLUSION: TRBAbs are found in 20% of hypothyroid patients with Hashimotos thyroiditis in assay using hTSHR-CHO cells, and they seem to play a role in the development of hypothyroidism in some patients with Hashimotos thyroiditis. TRBAbs of Hashimotos thyroiditis are different in epitope recognition from TRBAbs of primary myxedema.
Animals ; Antibodies* ; Cell Line ; Humans ; Hypothyroidism ; Immunoglobulin G ; Myxedema ; Rats ; Receptors, Thyrotropin* ; Thyroid Gland* ; Thyroiditis* ; Thyroiditis, Autoimmune

No abstract available.

The genetic status of cagA, vacA subtype, iceA1, and babA, and the relationship to gastroduodenal diseases were assessed in Helicobacter pylori isolates in Korea. Seventy-six strains of H. pylori were isolated from the antrum and the corpus of 41 adult patients (22 with peptic ulcer and 19 with gastritis). The cagA, iceA1, and babA genes were assessed by polymerase chain reaction and the vacA subtypes were determined by reverse hybridization-line probe assay. The positive rates of 349-bp cagA, 208-bp cagA, iceA1, and babA genes were 97.4%, 96.1%, 84.2%, and 36.1%, respectively. The vacA s1a, s1b, s1c, and s2 variants were detected in 11.8%, 3.9%, 80.4%, and 1.3%, respectively. m1 (78.9%) is more prevalent than m2 (5.3%). The most common vacA genotype was s1c/m1 (61.9%), and 14 isolates (18.4%) contained mixed vacA genotypes from a single biopsy specimen. Twenty-one (60%) of 35 patients were infected with more than two strains of different cagA, iceA1, babA, and vacA genotypes. None of cagA, iceA1, babA, and vacA s1/m1 were associated with peptic ulcer. In conclusion, most H. pylori isolates in Korea carry cagA, iceA1, and vacA s1c/m1 genes, and reside with multiple strains. These genes do not correlate with the peptic ulcer in the Korean patients.
Adult ; Aged ; Bacterial Proteins/*genetics ; Female ; Genotype ; Helicobacter pylori/*classification/genetics/pathogenicity ; Human ; Male ; Middle Age ; Peptic Ulcer/*etiology/microbiology

BACKGROUND: It has been known that most of thyroid stimulating antibodies (TSAbs) may interact with epitopes near N-terminal, and thyroid stimulation blocking antibodies (TSBAbs) near C-terminal on the extracellular domain of TSH receptor. However, many authors have reported different results about epitopes reacting with TSH receptor autoantibody (TRAb). TSBAbs inhibit thyroid stimulation of TSH and TSAbs at the receptor level. However, it has been reported that there are some TSBAbs which bind to the other sites, not TSH receptor, or block post-reeeptor process. These findings raise the possibility that TRAbs may be heterogeneous according to the mechanism of action. In order to investigate the heterogeneity of TRAb, we undertook immuno-precipitation using synthetic peptides of TSH receptor and measured TRAb activities by FRTL-5 cells and chimeric CHO cells. METHODS: We studied 102 patients with autoimmune thyroid disease (Graves disease 32, Hashimotos thyroiditis 29, atrophic thyroiditis 41) and 35 healthy persons. Three synthetic peptide fragments of TSH receptor were used to perform immunoprecipitation with serum or IgG of patients and healthy persons, TSAb and TSBAb activities were measured by FRTL-5 cells and CHO cells transfected with wild-type and 2 mutant TSH receptor cDNA (Mc2, Mc1+2). Mc2 and Mcl+2 were rnade to substitute amino acid residues of 90-165, 8-165 of the TSH receptor with corresponding residues of LH/CG receptor, respectively. RESULTS: Two out of 10 IgGs extracted from Graves disease and 2 out of 9 IgGs from atrophic thyroiditis had specific bidings over 0.84% in immunoprecipitation with peptide I (amino acid residue 35-50). Four out of 18 IgGs from Graves disease, 9 out of 41 IgGs from atrophic thyroiditis, and 6 out of 14 IgGs from Hashimotos thyroiditis had specific bidings over 0.84% in immunoprecipitation with peptide II (amino acid residue 317-332). Only 2 out of 10 IgGs from Graves disease had specific bidings over 0.84% in immunoprecipitation with peptide III (amino acid residue 341-358). When 10 IgGs extracted from Graves disease were reacted with wild-type, Mc2, and Mcl+2 CHO cells, 7 IgGs in wild-type and 4 IgGs in Mc2 had positive for TSAb activities. In 10 IgGs from atrophic thyroiditis, 5 in wild-type, 5 in Mc2, and 3 in Mcl+2 CHO cells had positive for TSBAb activities. In Hashimoto's thyroiditis, only 1 with hyperthyroidism had positive for TSAb activity in wild-type and 1 with hypothyroidism had positive for TSBAb activities in both of wild-type and Mc2 CHO cells. Therefore, patients with Graves disease were divided into at least 3 groups according to the TSAb activities measured by wild-type, Mc2, Mcl+ 2 CHO cells and TBII activities. And patients with atrophic thyroiditis were divided into at least 4 groups according to the TBII activities, TSBAb activities by wild-type, Mc2, Mcl+2 CHO cells and FRTL-5 cells. CONCLUSION: From these results, epitopes of TSH receptor reacting with TSAb or TSBAb in autoimmune thyroid disease may be scattered in the TSH receptor, although epitopes of TSAb tend to be near N-terminal and those of TSBAb near C-terminal. Graves disease or atrophic thyroiditis were divided into 3 or 4 groups according to the TBII and TRAb activities. Therefore, TRAb detected in autoimmune thyroid disease may be heterogenous.
Animals ; Antibodies, Blocking ; Autoantibodies* ; CHO Cells ; Cricetinae ; DNA, Complementary ; Epitopes ; Graves Disease ; Humans ; Hyperthyroidism ; Hypothyroidism ; Immunoglobulin G ; Immunoglobulins, Thyroid-Stimulating ; Immunoprecipitation ; Peptide Fragments ; Peptides ; Population Characteristics* ; Receptors, Thyrotropin* ; Thyroid Diseases* ; Thyroid Gland* ; Thyroiditis

No abstract available.
Humans ; Infant, Newborn* ; Mass Screening*

BACKGROUND: It has been widely accepted that the epitope(s) and/or functional characteristics of thyrotropin receptor antibodies (TSHRAb) from Graves' patients are heterogenous among patients. However, the clinical significance of such heterogeneity has not been systematically evaluated yet. We were to elucidate and find the clinical significance of heterogeneity for TSH receptor antibodies in Graves' disease. METHODS: We measured stimulating TSHRAb (TSAb) activities using CHO-hTSHR cells, FRTL-5 cells and chimeric receptor expressing cells (Mc1 + 2 and Mc2), specific blocking TSHRAb (TSBAb) activities using Mc2 cells and TBII activities using porcine thyroid membrane in 136 patients with untreated hyperthyroid Graves' disease. RESULTS: Based on various TSHRAb activities from each patient, the patients could be categorized into 7 subgroups by cluster analysis; 1) Group 1 (n = 41) was characterized by moderate TSAb activities both in CHO-hTSHR cells and in FRTL-5 cells, typical TSAb epitope, rare blocking antibodies and high TBII activities. 2) Group 2 (n = 16) was characterized by the presence of blocking TSHRAb in most patients, albeit the other characteristics were the same as those in Group 1. 3) Group 3 (n = 19) patients had low TSAb activities both in CHO-hTSHR cells and in FRTL-5 cells, seldom had blocking TSHRAb, but they had high TBII activities. 4) Group 4 (n = 30) could be categorized as 'mild disease' group, as they had low activities in all kinds of TSHRAb assay and had low antimicrosomal antibody activities. 5) Group 5 (n = 14) was characterized by moderate TSAb activities with atypical epitope(s), rare blocking TSHRAb and moderate TBII activities. 6) Group 6 (n = 10) patients had very high TSAb activities with typical epitopes, seldom blocking TSHRAb and low TBII activities. 7) Group 7 (n = 6) was characterized by very high TSAb activities with atypical epitopes and high TBII activities. Pretreatment serum thyroid hormone level was low only in group 4 patients compared to the other 6 groups (p < 0.05). The size of goiter was significantly larger in those in group 1 and group 3 (p < 0.05) compared to the other 5 groups. The prevalence of clinically significant ophthalmopathy was higher in group 2 patients than the other 6 groups (50% vs. 27.5%, p = 0.06). Among 6 kinds of TSHRAb activities, only the blocking TSHRAb activity was significantly associated with the presence of ophthalmopathy in multivariate analysis. CONCLUSION: These results suggest that the differences in epitopes for TSAb or the presence of blocking TSHRAb is not a major factor in determining the degree of thyrotoxicosis in Graves' disease. Although the pathogenic mechanism is not clear yet, we suggest that patients with ophthalmopathy have different TSHRAb repertoire from those without ophthalmopathy in Graves' disease.
Adolescent ; Adult ; Aged ; Cluster Analysis ; Comparative Study ; Female ; Graves' Disease/*classification/*immunology ; Human ; Immunoglobulins, Thyroid-Stimulating/*analysis/blood ; Logistic Models ; Male ; Middle Age ; Multivariate Analysis ; Receptors, Thyrotropin/analysis/*immunology ; Sensitivity and Specificity ; Severity of Illness Index

Recent molecular and physiological studies suggested that at least two H/K-ATPase isozymes are expressed in the rat kidney, and two distinct isoforms(HK alpha 2a, 2b) are resulted from alternative splicing of the 5-end of HK alpha 2 in distal colon by sequence analysis. Northern analysis and in situ hybridization(ISH) were carried out to analyze the expression of HK alpha 2a. and HK alpha 2b, mRNAs in rat kidney according to the changes of K-diet. Isoform specific 32P-labeled cDNA(for Northern) or digoxigenin labeled cRNA(for ISH) probes were used. Northern analysis demonstrated that HK a z. mRNA is abundantly expressed in normal(group 1: normal diet 2W) renal cortex, modestly in normal outer medulla, and weakly in normal inner medulla. The potassium-deprived rats(group 2: K-free diet 1W, and group 4: K-free diet 2W) expressed 40N lower levels in cortex and 2-4 fold higher levels of HKalpha 2a. mRNA in outer and inner medulla compared to normal rat. The potassium loading rats after potassium-deprivation(group 3: normal diet 1W after K-free diet 1W, and group 5: normal diet 1W after K-free diet 2W showed almost normal levels of HK alpha 2a. mRNA. HK alpha 2b, mRNA was not detected in any tissues of groups. By ISH, mRNA for HK alpha 2, was detected in the thick ascending limb, distal convoluted tubule, and the entire collecting duct. All groups exhibited comparable cellular patterns of labeling. Signal intensity of group 2 and 4 was less in cortical collecting duct(CCD), especially principal cells and much higher in the inner stripe of the outer medullary collecting duct(OMCDi) and the proximal inner medullary collecting duct(IMCD) compared to group 1. Group 3 and 5 exhibited signal intensity of group l. These results indicate that chronic hypokalemia enhances expression of HK alpha 2a, gene in OMCDi and proximal IMCD, decreases in CCD, and restores at normal levels in potassium-loading after potassium-deprivation and suggest that this isoform plays an important role in potassium balance by these segments accordiog to the changes of K-diet.
Alternative Splicing ; Animals ; Colon ; Diet* ; Digoxigenin ; Extremities ; Hypokalemia ; In Situ Hybridization ; Isoenzymes ; Kidney ; Potassium ; Rats ; RNA, Messenger ; Sequence Analysis

BACKGROUND: It is important to recognize the independent prognostic factors of papillary carcinoma of thyroid in therapeutic and follow up planning. However, its good prognosis and its low prevalence make it difficult to analyze prognostic factors determining patients survival in a relatively short period of time. We retrospectively investigated the independent prognostic factors for determining disease recurrence after surgery which, in adults, are known to be closely related to the prognosis of cancer. METHOD: We retrospectively reviewed the clinical records of 456 patients (male 70, female 381, unknown 5, mean age of 43.9+-12.9 years) who had visited the thyroid clinic in Seoul National University Hospital and analyzed the data with statistical software program. RESULTS: 1) At initial visit, chief complaint of the patients was abnormal neck mass in 90 percent. 2) In preoperative thyroid scan study, 82% showed cold area and 16% showed diffuse enlargement. 3) Of the 50 recurrent cases after surgery (11.5% of the total cases), 39 cases (78%) had recurred disease in neck area and 11 cases (22%) had recurrences at distant sites. (Lung 9 cases, Brain I case, Mediastinum 1case) 4) Statistically significant risk factors for recurrence after surgery were male sex, size of tumor (above 4.5cm in this study), extrathyroidal invasion of cancer, involvement of resection margin and no remnant ablation of thyroid tissue using radioiodine. CONCLUSION: Through retrospective study, we presented some clinical characteristics of papillary thyroid cancer in Korea and independent risk factors of cancer recurrences after surgery.
Adult ; Brain ; Carcinoma, Papillary ; Female ; Follow-Up Studies ; Humans ; Korea ; Male ; Mediastinum ; Neck ; Prevalence ; Prognosis ; Recurrence* ; Retrospective Studies ; Risk Factors ; Seoul ; Thyroid Gland* ; Thyroid Neoplasms*