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L J DeGroot

Publications and source records attributed to L J DeGroot.

At least 145 records · Page 8Linked to original sources
PubMed1987

Thyroid hormone receptors in a human hepatoma cell line: multiple receptor forms on isoelectric focusing.

The nuclear thyroid hormone receptors isolated from cultured human hepatoma cells (Hep G2) were characterized and compared with those from cultured human fibroblasts and rat liver. The Hep G2 nuclear thyroid hormone receptors had an affinity constant (Ka) of 2.1 X 10(10) M-1 and maximal binding capacity (MBC) of 21.0 fmol/100 micrograms DNA for T3 in assays performed on isolated nuclei. 16% of nuclear receptors were released into the media during incubation and had the same Ka. Salt-extracted receptors had a Ka of 1.8 X 10(10) M-1 and MBC of 0.1 pmol/mg protein for T3. Density gradient sedimentation and gel filtration chromatography revealed a sedimentation coefficient of 3.4 S and Stokes radius of 34 A. From these values, a molecular weight of 49,000 and total frictional ratio (f/f0) of 1.4 were calculated, suggesting an asymmetrical shape of the receptor molecule. Heat inactivation occurred with t1/2 of 28.1, 18.0, and 7.9 min at 38, 43, 45 degrees C, respectively. Isoelectric focusing (IEF) of Hep G2 nuclear receptors demonstrated T3 binding proteins at pH 5.3-5.5, 5.7, and 5.9. Evidence that these are nuclear thyroid hormone receptors includes the following: Triiodothyroacetic acid was the most potent competitor of [125I]T3 binding to these proteins followed by L-T3, and L-T4. Cytosolic protein, human serum, and fetal calf serum failed to show the same T3 binding proteins. Ka of these proteins measured by T3 displacement was 1.1-3.2 X 10(9) M-1. Human fibroblast nuclear extract showed similar T3 binding pattern in IEF, except for a slight difference in pI of an acidic band.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding, Competitive↗
PubMed1987

Purification and characterization of rat liver nuclear thyroid hormone receptors.

Nuclear thyroid hormone receptor was purified to 904 pmol of L-3,5,3'-triiodothyronine (T3) binding capacity per mg of protein with 2.5-5.2% recovery by sequentially using hydroxylapatite column chromatography, ammonium sulfate precipitation, Sephadex G-150 gel filtration, DNA-cellulose column chromatography, DEAE-Sephadex column chromatography, and heparin-Sepharose column chromatography. Assuming that one T3 molecule binds to the 49,000-Da unit of the receptor, we reproducibly obtained 6.4-14.7 micrograms of receptor protein with 4.2-4.9% purity from 4-5 kg of rat liver. Elution of receptor from the heparin-Sepharose column was performed using 10 mM pyridoxal 5'-phosphate, which was observed to diminish binding of receptor to heparin-Sepharose or DNA-cellulose. This effect was specific for pyridoxal 5'-phosphate, since related compounds were not effective. Purified receptor bound T3 with high affinity (6.0 X 10(9) liter/mol), and the order of affinity of iodothyronine analogues to purified receptor was identical to that observed with crude receptor preparations [3,5,3'-triiodothyroacetic acid greater than L-T3 greater than D-3,5,3'-triiodothyronine (D-T3) greater than L-thyroxine greater than D-thyroxine]. Purified receptor had a sedimentation coefficient of 3.4 S, Stokes radius of 34 A, and calculated molecular mass of 49,000. Among several bands identified by silver staining after electrophoresis in NaDodSO4/polyacrylamide gels, one 49,000-Da protein showed photoaffinity labeling with [125I]thyroxine that was displaceable with excess unlabeled T3. The tryptic fragment and endogenous proteinase-digested fragment of the affinity-labeled receptor showed saturable binding in 27,000-Da and 36,000-Da peptides, respectively. These molecular masses are in agreement with estimates from gel filtration and gradient sedimentation, indicating that affinity labeling occurred at the hormone binding domain of nuclear thyroid hormone receptor. This procedure reproducibly provides classical native rat liver T3 nuclear receptor in useful quantity and purity and of the highest specific activity so far reported.

Animals↗
PubMed1987

Characterization and isolation of thyroid microsomal antigen.

We investigated the structure of the 107-kD thyroid protein recognized as microsomal antigen. Solubilized microsomes were incubated with affinity gels consisting of IgG, from thyroiditis patients or controls, linked to Reacti-gel. Eluates were analyzed by SDS polyacrylamide electrophoresis and Western blot. 107- and 101-kD proteins were augmented in eluates from gels containing patient IgG and had microsomal antigenicity. In a Western blot of microsomes run under unreduced conditions, poorly defined large proteins were identified by antibody. When eluted electrophoretically and reanalyzed in reducing conditions, they demonstrated the 107-kD antigen. The 107-kD protein identified in reducing conditions was extracted and reanalyzed under nonreducing conditions. Large molecular mass proteins were then observed. On two-dimensional electrophoresis, a 107-kD antigen was isolated with isoelectric point of 7.0. The microsomal antigen may be complexes or multimers of a 107-kD peptide with isoelectric point of 7.0.

Antigens↗
PubMed1987

Isolation of a complementary DNA clone for thyroid microsomal antigen. Homology with the gene for thyroid peroxidase.

The thyroid microsomal antigen (MSA) in autoimmune thyroid disease is a protein of approximately 107 kD. We screened a human thyroid cDNA library constructed in the expression vector lambda gt11 with anti-107-kD monoclonal antibodies. Of five clones obtained, the recombinant beta-galactosidase fusion protein from one clone (PM-5) was confirmed to react with the monoclonal antiserum. The complementary DNA (cDNA) insert from PM-5 (0.8 kb) was used as a probe on Northern blot analysis to estimate the size of the mRNA coding for the MSA. The 2.9-kb messenger RNA (mRNA) species observed was the same size as that coding for human thyroid peroxidase (TPO). The probe did not bind to human liver mRNA, indicating the thyroid-specific nature of the PM-5-related mRNA. The nucleotide sequence of PM-5 (842 bp) was determined and consisted of a single open reading frame. Comparison of the nucleotide sequence of PM-5 with that presently available for pig TPO indicates 84% homology. In conclusion, a cDNA clone representing part of the microsomal antigen has been isolated. Sequence homology with porcine TPO, as well as identity in the size of the mRNA species for both the microsomal antigen and TPO, indicate that the microsomal antigen is, at least in part, TPO.

Amino Acid Sequence↗
PubMed1987

Modification of deoxyribonucleic acid-thyroid hormone receptor interaction by histones.

The effects of histones on receptor-DNA interaction were examined using an in vitro DNA-cellulose-binding assay of [125I]T3 receptor. H1 histones bound to DNA-cellulose strongly inhibited binding of receptor to DNA-cellulose. DNA-cellulose column chromatography showed that 80% of T3-binding activity attached to DNA-cellulose could be eluted using 1 mg H1 histone/ml at low ionic strength. The potent inhibitory activity of H1 histones on receptor-DNA binding was reversed by removal of H1 histones from DNA-cellulose or by removal of H1 histones from receptor. The interaction was specific for the DNA-binding activity of receptor, since H1 histones inhibit neither T3-binding activity nor core histone-binding activity of receptor. In contrast, low concentrations of core histones enhanced binding of T3 receptor to DNA-cellulose. This effect was also seen when DNA-cellulose was treated with core histones and then deprived of free core histones. The enhancement was reversed by removal of core histones bound to DNA-cellulose. A tryptic fragment of the receptor, which lost DNA-binding activity but retained hormone and core histone-binding activities, was capable of binding to DNA-cellulose in the presence of core histones. These data suggest that enhanced binding of receptor to DNA-cellulose by core histones is mediated through the core histone-binding activity of receptor. A heat-labile protein factor in nuclear extracts (possibly receptor itself) also enhanced receptor binding to DNA-cellulose. Our data suggest important roles of histones in the organization of nuclear thyroid hormone receptors in chromatin. Since H1 histone participates in condensation of chromatin and is enriched in transcriptionally inactive chromatin, inhibition of DNA-binding activity of receptor by H1 histone bound to DNA could explain the preferential binding of nuclear thyroid hormone receptor to transcriptionally active chromatin.

Animals↗
PubMed1987

Antibodies against denatured and reduced thyroid microsomal antigen in autoimmune thyroid disease.

Different antigenic determinants on thyroid microsomal antigen (MAg) could induce different antibodies, which, in turn, could have differing importance in the pathogenesis of autoimmune thyroid disease. In this study we demonstrated three types of microsomal antibodies (MAbs) present in serum of patients with autoimmune thyroid disease by Western blot analysis. These MAbs reacted with native, denatured, and denatured and reduced MAg. Methods for detecting the MAbs were developed, and the clinical significance of these Abs was analyzed in 197 patients with thyroid disorders. For measurement of Ab against native and denatured MAg, an enzyme-linked immunosorbent assay, in which wells were coated with untreated or sodium dodecyl sulfate-denatured MAg, was used. For measurement of Ab against denatured and reduced MAg, 6% SDS-polyacrylamide gel electrophoresis followed by Western blot analysis was used. Native MAb enzyme-linked immunosorbent assay values correlated highly with microsomal hemagglutination titers, but patients with high titers of Ab against native MAg did not necessarily have Ab against denatured MAg or reduced MAg. Abs against denatured MAg or denatured and reduced MAg were found in patients with Hashimoto's disease (28.8% and 13.6%) and Graves' disease (22.4% and 11.2%). The percentage of patients with positive denatured and reduced MAb was higher in Graves' disease patients who had a longer sick interval or who developed hypothyroidism after radioiodine treatment. This study provides the first clear demonstration that MAbs are heterogeneous. The data suggest that antibodies against denatured or denatured and reduced MAg may be related to destruction of the thyroid gland.

Autoantibodies↗
PubMed1987

Molecular cloning of the complementary deoxyribonucleic acid for human thyroid peroxidase.

Five overlapping cDNA clones representing the entire mRNA for human thyroid peroxidase (TPO) have been isolated from a human Graves' thyroid cDNA library. The cDNA sequence has been determined. Human TPO cDNA contains 3060 bases from the start of transcription to the beginning of the poly (A) tail at the 3'-end. The derived amino acid sequence of human TPO consists of 933 amino acids with a mol wt of 102,937. The derived amino acid sequence contains five potential glycosylation sites (Asn-X-Ser/Thr), a probable transmembrane signal peptide sequence at the amino terminus, and a hydrophobic putative membrane-spanning region beginning 85 amino acid residues from the carboxyl terminal end. Comparison of the human TPO amino acid sequence to that of pig TPO shows strong homology extending from the amino terminus to within 44 amino acid residues of the carboxyl-terminus.

Amino Acid Sequence↗
PubMed1987

Spontaneous in vitro production of thyroglobulin specific helper factor in patients with autoimmune thyroid diseases.

We have investigated regulatory cell abnormalities in patients with autoimmune thyroid diseases through in vitro thyroglobulin (Tg) specific helper factor production. Helper activities were measured in the mice spleen cell assays, using hapten-carrier system. Peripheral blood leukocytes (PBL) from 5 out of 6 patients produced the Tg specific helper factor not only when they were stimulated in vitro with Tg but also when they were not stimulated. On the contrary, normal PBL did not produce the factor when they were not stimulated in vitro. However, 3 out of 5 normal PBL did produce the factor when they were stimulated in vitro with Tg. From several lines of studies, this helper factor was specific for Tg in its production and action. From these results it is concluded that (a) Tg specific helper T cells are already activated in vivo in patients with autoimmune thyroid diseases, indicating the presence of immunoregulatory abnormalities in these patients, and that (b) normal PBL have a potential to produce this helper factor when they are appropriately stimulated.

Biological Assay↗
PubMed1987

HLA immunogenetic heterogeneity in black American patients with Graves' disease.

Seventy-three American black patients with Graves' disease were typed for HLA-A, HLA-B, and HLA-DR antigens. There was a slight increase in HLA-DRw6 antigen frequency compared with 238 normal American black controls, but this was not significant after correction for the number of antigens tested. A significant increase in HLA-DR4 and HLA-DRw6 frequency was found in a subgroup of patients with exophthalmos (22.9% and 29.2% compared with 7.6% and 10.1% of normal black controls). There was a significant increase in HLA-DRw6 in a subgroup of patients who were thyroid antibody-positive (26.0%). The increment in HLA-DRw6 was higher in 32 patients who had both exophthalmos and who were antibody positive (37.5%). A significant increase in HLA-DR5 was found in a subgroup of patients who did not have exophthalmos and who were antibody-negative (83.3%). Our findings support previous evidence for immunogenetic heterogeneity in patients with toxic Graves' disease. In American blacks HLA-DRw6 is in some way associated with the disease in contrast to the well-recognized HLA-DR3 association in whites.

Black or African American↗
PubMed1987

Physiochemical factors influencing the stability of solubilized liver nuclear receptor-3,5,3'-triiodothyronine complex in vitro.

Nuclear acidic protein fraction containing saturable receptors for thyroid hormones was prepared from purified rat liver nuclei by extraction with 0.3 mol l-1 KCl. The inhibitory effect of an increasing ionic strength on the 3,5,3'-triiodothyronine (T3) binding to receptors at pH 8.0 for 2 h and tested up to 2.0 mol l-1 KCl was confirmed. The nuclear receptor-T3 complex was found resistant to the effect of a higher ionic strength up to 1 mol l-1 KCl when subsequently incubated at pH 8.0 for 1 h. In addition, a rapid dissociation of the receptor-T3 complex at alkaline pH in the order of 10.5 greater than 10.0 greater than 9.0 greater than 8.0 was re-examined. The effect of ATP ranging from 0.05 mmol l-1 to 1.0 mmol l-1 on the stability of the receptor-T3 complex at pH 8.0 was also studied. The experiments were conducted in the presence of 25 mmol l-1 Na2HPO4 in order to suppress the effect of the phosphate groups of ATP molecule by inorganic phosphate. The data confirmed that the concentration of ATP starting from 0.05 mmol l-1 may cause the dissociation of nuclear receptor-T3 complex in vitro.

Adenosine Triphosphate↗
PubMed1986

Separation of DNA binding domain from hormone and core histone binding domains by trypsin digestion of rat liver nuclear thyroid hormone receptor.

Rat liver nuclear thyroid hormone receptor was subjected to limited trypsin digestion, and the tryptic fragment of the 3,5,3'-triiodo-L-thyronine (T3)-receptor complex was characterized. Rat liver nuclear thyroid hormone receptor is an asymmetrical protein with Stokes radius of 34 A, sedimentation coefficient of 3.4 S, and molecular weight of 49,000. A globular T3-receptor complex with Stokes radius of 22 A, sedimentation coefficient of 2.8 S, and molecular weight of 26,000 was obtained by tryptic digestion. This fragment had no DNA binding activity, whereas undigested receptor showed significant DNA binding activity. Addition of undigested receptor to the tryptic fragment did not restore DNA binding activity of digested receptor, nor did mixing inhibit DNA binding activity of undigested receptor complex. Undigested receptor bound to core histones, and this activity was stronger than with other proteins tested (H1 histone, cytochrome c, and ovalbumin). The tryptic fragment of receptor maintained core histone binding activity comparable to that of undigested receptor. The tryptic fragment had affinity for T3 comparable to undigested receptor as assessed by Scatchard analysis and the same rate for dissociation of [125I]T3 from receptor. The tryptic fragment of the T3-receptor complex was more stable than undigested receptor at 43 degrees C. Digestion of receptor unoccupied by T3 caused a significantly larger loss of T3 binding capacity than did digestion of T3-occupied receptor, suggesting a protective effect of T3 on a second trypsin-sensitive site on the receptor, which, when cut, destroys T3 binding activity.

Animals↗
PubMed1986

Nuclear thyroid hormone receptors in cultured human fibroblasts: improved method of isolation, partial characterization, and interaction with chromatin.

In order to characterize the nuclear thyroid hormone receptors in human tissue, an improved method for isolation of nuclei from cultured human fibroblasts was developed. This method provided nuclei with a protein/DNA ratio of 2.8 and recovery of 42%. The purity of nuclei was verified by phase contrast and electron microscopy, which showed normal appearance of chromatin structure. Nuclear binding assay was performed by incubation of whole cells at 37 degrees C or isolated nuclei at 22 degrees C with L-triiodothyronine (T3). In both cases, an affinity constant (Ka) of 2.0-3.0 X 10(10) M-1 and an average binding capacity of 41 femtomoles of T3/100 micrograms DNA (3,100 binding sites/nucleus) were obtained. During incubation of the nuclei, 13% to 16% of receptors that had an identical Ka was released into the medium. Salt extraction recovered 85% to 90% of the receptors, which had a Ka of 4.5 X 10(10) M-1 and the capacity of 0.13 pmol of T3/mg protein. The Ka fo. L-thyroxine (T4) was seven to 18 times lower than that for T3, but the capacity was the same in isolated nuclei, receptors released during incubation of nuclei, and in salt-extracted receptors. Of the iodothyronines examined, affinity for triiodothyroacetic acid was the highest, followed by L-T3, D-T3, L-T4. Isokinetic glycerol gradient analysis revealed that salt-extracted receptors had a sedimentation coefficient of 3.4 S, whereas micrococcal nuclease digested receptors showed two major (6.0 to 6.5 and 12.5 S), and two minor (17 and 19 S) peaks. These results were virtually identical to those obtained with rat liver nuclei analyzed in parallel studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding Sites↗
PubMed1986

Hürthle cell tumors: a twenty-five-year experience.

During a 25-year period (1959 through 1983), 54 patients with Hürthle cell tumors were treated and monitored at the University of Chicago Medical Center. Thirty percent were men and 70% were women; mean age at diagnosis was 46.7 +/- 13.2 years (range: 19 to 69 years). Tumors were grouped into three categories at the time of initial diagnosis: group 1, grossly malignant (four patients, or 7.5%); group 2, intermediate (partial capsular and/or subcapsular vascular invasion) (10 patients, or 18.5%); and group 3, benign appearing (40 patients, or 74%). Twenty-one (39%) of the patients had a history of low-dose, external radiation to the head and neck in childhood (including three of four grossly malignant lesions). A separate non-Hürthle cell thyroid carcinoma was found within the thyroid gland in 22 (50%) of the patients--79% were papillary and 21% were follicular carcinomas. In half of these, there was a history of childhood irradiation. During a mean follow-up period of 8.4 years (range, 22 days to 35 years), three additional Hürthle cell tumors were recognized as malignant after metastases were discovered--two were originally classified as intermediate lesions and one was in the benign-appearing group. Thus, seven of 54 of our patients (13%) had Hürthle cell carcinomas. One of the seven patients died of widespread metastases after 35 years, and the other six are currently free of disease. We believe that therapy of these lesions should be individualized. Total thyroid ablation (surgical procedure followed by radioiodine therapy) is appropriate for frankly malignant Hürthle cell cancers, for all Hürthle cell tumors occurring in patients who received low-dose childhood irradiation, for associated papillary or follicular carcinomas, and in those patients who exhibit partial capsular or subcapsular vascular invasion. On the other hand, single, well-encapsulated, benign-appearing Hürthle cell tumors may be treated by lobectomy and careful follow-up, since the chance that they will later exhibit malignant behavior is low (2.5% in our series and 1.5% among patients described in the recent literature).

Adenoma↗
PubMed1986

Graves' disease and thyroid cancer.

The relationship between Graves' disease or its therapy and carcinoma of the thyroid remains uncertain. We studied 20 patients found to have thyroid cancer in glands previously treated for Graves' disease between 1961 and 1986 at the University of Chicago Medical Center. Sixteen (80%) occurred in women and four (20%) occurred in men. The mean age at operation was 37 years (range, 19 to 69 years) and did not differ by sex. Fifteen of the 20 cancers (75%) were papillary while five (25%) were follicular. Six individuals (30%) had a history of external radiation to the head and neck as an infant, child, or young adult. Two others had received radioiodine (RAI) therapy for Graves' disease 1 and 19 years earlier. Patients were divided into three groups: group I: four patients (20%) had a neck mass 4, 14, 20, and 41 years after having had a subtotal thyroidectomy (STT) for Graves' disease; three of four had a history of external irradiation therapy. These tumors behaved aggressively, resulting in the death of two of the four patients. group II: 11 patients (55%) had diffusely enlarge toxic goiters without a nodule. A carcinoma was diagnosed intraoperatively on frozen section in only two of these patients. The others received STT. After recognition on permanent section, those carcinomas that were 4 mm or greater in diameter received postoperative RAI. One recurrence occurred and was treated successfully with further RAI. group III: Five patients (25%) had Graves' disease and a palpable thyroid nodule. None of them had had a prior thyroidectomy for Graves' disease, as in group 1. Thyroid carcinoma was diagnosed in all patients preoperatively or intraoperatively, and a total thyroidectomy was performed. Each patient is alive and well with a mean follow-up of 5 years. Between 1971 and 1981, 194 patients had surgery for Graves' disease, and 10 (5.2%) were found to have an associated carcinoma; six patients (3.1% of the total) did not have a nodule or any other suspicion of malignancy before surgery. During the same time, 303 patients received RAI therapy for Graves' disease and one (0.3%) has subsequently developed thyroid carcinoma. Thyroid cancer associated with Graves' disease is found more commonly in surgically treated patients than in patients after RAI therapy. The greatest risk factor in our patients was previous external radiation to the head and neck. Such individuals should be treated with total thyroid ablation rather than the usual STT, since they are at risk of developing aggressive thyroid cancers if thyroid remnants are left.

Adult↗
PubMed1985

Association of differentiated thyroid carcinoma with HLA-DR7.

Seventy-four American white thyroid cancer patients were typed for HLA-A, B, and DR antigens. A significant increase in HLA-DR7 was found in the nonradiation-associated thyroid cancer patients (42.5%, 20/47 cases), compared to 22.8% of 979 normal controls. The association is stronger in the follicular and mixed papillary-follicular subgroup (52.0%, 13/25 cases, P corrected less than 0.01). The occurrence of various malignancies in family members was found in 57.9% of HLA-DR7 positive patients, versus 20% of HLA-DR7 negative patients, in a retrospective record review. Although the frequency of HLA-DR7 was not increased in the radiation-associated thyroid cancer patients (22.2%, 6/27 cases), the interval from the irradiation date to the onset date of thyroid cancer was shorter in HLA-DR7 positive cases (17.3 +/- 6.2 years) than in HLA-DR7 negative patients (29.4 +/- 11.5 years). This data suggest that HLA-DR7 is associated with and may influence development of thyroid cancer.

Adenocarcinoma↗
PubMed1985

Identification of a thyroid microsomal antigen by Western blot and immunoprecipitation.

The molecular identity of the thyroid microsomal antigen was investigated by Western blot and immunoprecipitation using sera from eight patients with autoimmune thyroid disease. All sera had high microsomal antibody titers by hemagglutination test and ELISA; only one had thyroglobulin (Tg) antibody. In an immunoprecipitation study using Triton X-100-solubilized 125I-labeled microsomes, all of the sera precipitated a 107K protein. However, this 107K protein was visualized by only three patients' sera by Western blot analysis under reducing conditions. In Western blots run under nonreducing conditions, four sera, including the three sera mentioned above, recognized poorly defined large mol wt proteins. One serum which also had Tg antibody recognized additional bands by both immunoprecipitation and Western blot. Preincubation of serum with 100 micrograms/ml Tg or 100 mU/ml bovine TSH did not alter the binding of antibody to 107K protein in Western blot analysis. The 107K protein was not visualized in Western blots of liver and kidney microsomes using patients' sera. Experiments were performed to characterize the difference between one serum (no. 1) which visualized the 107K band in Western blots and a serum (no. 4) which did not. Reactivity at different dilutions was compared by ELISA and Western blot studies. Serum 1 recognized the 107K protein even at a 1:3200 dilution, which gave a lower optical density value than a 1:200 dilution of serum 4 in ELISA, while serum 4 failed to recognize the 107K band at any dilution. An affinity gel prepared from serum 4 immunoglobulin G linked to agarose [Reacti-Gel (6 X)] removed from Triton X-100-solubilized microsomes the 107K protein which patient 1 serum recognized by Western blot analysis under reducing conditions. The data indicate that serum 4 can recognize the 107K protein under nondenaturing conditions. These data indicate that the 107K protein described here is the microsomal antigen, and that may associate with another protein or maintain a unique conformation due to disulfide bonds present in the native state. In addition, the 107K protein includes at least two antigen epitopes and different patients have different antibodies or different populations of antibodies against these epitopes.

Antigens↗
PubMed1985

Studies on thyroglobulin-specific suppressor T cell function in autoimmune thyroid disease.

T cell regulation of the generation of thyroglobulin plaque-forming cells (Tg PFC) and protein A plaque-forming cells (Prot A PFC) was investigated using lymphocytes from patients with autoimmune thyroid disease. T and B cell mixed cultures (T-B MC) were carried out without mitogenic or antigenic stimulation to identify physiological T cell effects in the system. Tg PFC were found in 8 (44%) of 18 patients who had high titers of thyroglobulin antibody in their sera. Tg-specific and nonspecific immunoregulation by T cells from patients and normal subjects was studied using B cells from these eight patients in the T-B MC system. Remarkably lower values of Tg PFC induction compared to Prot A PFC induction were found after T cell addition. Normal T cells inhibited Tg PFC induction, but patient T cells did not, while the same extent of helper effects were found on Prot A PFC induction by the addition of patient and normal T cells. Irradiation (1500 rads) of T cells from patients and normal subjects significantly enhanced both TgPFC and Prot A PFC induction. Thus, Tg-specific suppressor T cells are present in all normal subjects as part of the radiosensitive suppressor T cell subset. The increase in Tg-PFC caused by irradiation-induced inhibition of Tg-specific suppressor T cell function was significantly greater in normal subjects than in patients. Histamine type 2 receptor-bearing T cells inhibited Prot A PFC induction, but not Tg PFC induction, in the autologous T-B MC system. No Tg PFC were induced from normal B cells in any combination with untreated T cells, irradiated T cells, or histamine type 2 receptor-negative T cells from patients or normal subjects. These data indicate that in vitro Tg-specific T cell regulation can be studied in the T-B MC system by using B cells from patients with autoimmune thyroid disease with high Tg antibody titers in their sera. Tg-specific suppressor T cells appear to be present in all individuals and to be involved in the regulation of Tg antibody production. The lower activity of Tg-specific suppressor T cells in patients compared to that in normal subjects may be related to Tg antibody production in vivo. This abnormality, however, is heterogeneous and is not a complete but, rather, is a relative defect of Tg-specific suppressor T cells.

Adolescent↗