Thyroid®

B lymphocytes that infiltrate the thyroid (Thy-B cells) in Graves' patients appear to be implicated in the pathophysiology of this disorder. The goal of the present study was to examine the nature of these Thy-B cells. To this end, Thy-B lymphocytes were isolated from surgical thyroidal samples, and their phenotype was determined by using mouse monoclonal antibodies (mAb) directed against a wide variety of surface markers, followed by flow cytometry multicolor analysis. The results show that most Thy-B cells (≅60%) exhibited IgM⁺ IgD^{low to-} surface immunoglobulin (Ig) profile, whereas the minor cell fraction (≅30%) consisted of switched IgG⁺ memory B lymphocytes. Thy-B cells expressed low levels of CD5, CD23, and CD62L, which distinguished them from the resting B-cell pool, the major B-cell subset in the blood. In addition, they lacked CD38, CD10, and CD71, characteristic molecules for the germinal center B lymphocytes. In addition, Thy-B lymphocytes showed peculiar patterns both of adhesion molecules (CD62L^-, CD44^intermediate), and of activation molecules (CD69⁺, CD80⁺, and, in part, CD95⁺). Taken together, these results suggest that the Thy-B lymphocyte subset consists of a combination of IgM⁺ B cells resembling marginal zone B lymphocytes, and isotype-switched memory B cells.

Autoimmune thyroid diseases feature prominent cellular infiltration of the thyroid gland as well as autoantibody production to thyroid antigens. The most common assay to evaluate cell-mediated immunity is based on incorporation of tritiated thymidine into proliferating T cells after stimulation by the test antigens. In the past, cell proliferation assays of thyroglobulin (Tg) using peripheral blood mononuclear cells (PBMC) of individuals with autoimmune thyroid diseases required large quantities of blood and specialized separation techniques, and have not yielded high counts or high stimulation indices. We therefore developed a proliferation assay using less than 5 mL of whole blood and compared proliferation of cells in whole blood to that using PBMCs separated by density gradient centrifugation. We also determined if responses could be enhanced by addition of interleukin-2 (IL-2) to the cultures. We found that an IL-2–stimulated proliferation assay to Tg using diluted whole blood is superior to the separated cell assay in detecting Tg-specific T-cell proliferation in autoimmune thyroid disease patients. Further refinement of this technique and larger trials may confirm its value for clinical investigation and special diagnostic applications.

We studied the regulation of the expression of vascular endothelial growth factor (VEGF) by TSH in monolayer cultures of rat thyroid FRTL-5 cell lines. The VEGF mRNA synthesis was significantly inhibited by TSH as well as dibutyryl cyclic adenosine monophosphate (cAMP) in a dose-dependent manner in FRTL-5 cells. This observation is contrary to previously published results using the thyroid follicular culture system. Our results suggest that the direct effect of TSH/cAMP is to inhibit the VEGF synthesis in monolayer cells.

Myoglobin (Mb) gene expression, Citrate Synthase (CS) and Succinate Dehydrogenase (SDH) activities of Soleus (S) and Extensorum Digitalis Longus (EDL) muscles were studied in intact, thyroidectomized and T3-treated (25ug/100g, BW, ip, 15 days) rats. The fiber type composition of S muscle was also evaluated and used as control of the T₃-induced effects. In the S muscle, the T₃ treatment increased the Mb mRNA and protein expression, as well as the CS and SDH activity. These changes occurred parallel to the expected increase in type II (fast) and decrease in type I (slow)-fibers in S muscle. In the hypothyroid state, the Mb mRNA was decreased, while the Mb expression and CS activity tended to decrease. In contrast the SDH activity was increased, probably due to the enhanced motor activity that occurs as a short-term response to the hypothermia induced by hypothyroidism. In the EDL, the alterations were milder than those in S muscle in both thyroid states. These findings show that Mb gene expression is induced by T₃. This is concomitant with the enhancement of Krebs Cycle enzyme activities and provides additional evidence that thyroid hormone increases the aerobic potential of skeletal muscles, as well as the speed of muscle contraction.

We describe Japanese siblings with resistance to thyrotropin (TSH) who are compound heterozygotes for two novel mutations in the TSH receptor gene. The affected siblings had increased serum TSH, normal serum thyroid hormones, and normal positioned but slightly hypoplastic thyroid glands. The mutated paternal allele has the substitution of His (CAC) in place of Arg (CGC) at codon 450 (R450H) of the TSH receptor. The mutated maternal allele has the substitution of Ser (AGT) in place of Gly (GGT) at codon 498 (G498S) of the TSH receptor. COS-7 cells transfected with the R450H mutant exhibited a slightly decreased TSH binding and a slightly decreased cyclic adenosine monophosphate (cAMP) response to TSH, whereas cells transfected with the G498S mutant exhibited a markedly decreased TSH binding and a markedly decreased cAMP response to TSH. Flow immunocytofluorometry analysis demonstrated that the G498S mutant resulted in extremely low expression at the cell surface as compared with the wild type receptor and the R450H mutant, in spite of a normal intracellular synthesis. The present cases are the first Japanese patients with TSH resistance in whom mutations in the TSH receptor gene have been identified. These novel mutations may contribute to understanding of the structure-function relationship of the TSH receptor.

Oral iodinated radiographic contrast agents such as ipodate and iopanoic acid form an important part of the armamentarium used to treat hyperthyroidism. They rapidly and dramatically reduce serum triiodothyronine(T₃) levels by inhibiting conversion of thyroxine (T₄) to T₃ in the periphery and by blocking secretion from the thyroid. Potential risks from the large iodine load resulting from their use limit their widespread applicability. In addition, they are ineffective when used alone on a long-term basis. However, these agents may be especially useful in treating thyrotoxic patients preoperatively, in neonatal Graves' disease, in massive levothyroxine ingestion, and when other conventional antithyroid drugs are unsuccessful or contraindicated.

Conventional two-dimensional (2-D) ultrasound is the standard method for the investigation of thyroid morphology. Volume calculations need model assumptions and are observer dependent. The present study performed with a commercially available three-dimensional (3-D) system Freescan added to a conventional ultrasound scanner compares the accuracy of conventional thyroid volumetry to several methods of 3-D volume determination. In vitro measurements were performed on thyroid phantoms with known volumes. The standard deviation of the normalized differences was 8.0% (3-D segmentation) and 10.5% (conventional). For the accuracy of volume determination in human thyroids we performed a postmortem study. The thyroid volume was calculated conventionally by the ellipsoid model and by two 3-D methods (segmentation and the newly developed multiplanar volume approximation). The reference volume was determined after resection by submersion. The standard deviation of the normalized differences was 26.9% for the conventional method, 9.7% for 3-D segmentation and 11.5% for the multiplanar volume approximation, showing significant better results for both 3-D methods and no significant difference between the 3-D methods. The 3-D system, therefore, achieves a better accuracy for thyroid volumetry than the conventional volumetry using planar images. In addition, the 3-D images are stored electronically and can be used for follow-up studies.

Antithyroid drugs have been reported to reduce the expression of HLA-DR in thyrocytes in Graves' disease, but only circumstantial evidence has been provided about their in vivo immunologic effects. This present study was designed to examine the in vivo immunologic effect of antithyroid drugs on thyroid follicular cells. The study was conducted on 25 patients who had Graves' disease in remission or in activity and who were or were not receiving treatment (7 in overt thyrotoxicosis, 6 patients in remission, and 12 patients under medication). HLA-DR expression in thyroid biopsies was verified by immunohistochemistry. The follicular cells of all patients in overt thyrotoxicosis expressed HLA-DR whereas those of patients in remission were negative for HLA-DR. HLA-DR was also not expressed in all patients under medication, but this did not correlate with the clinical evolution after thyroid drug withdrawal. In conclusion, antithyroid drugs inhibit follicular cell HLA-DR expression in Graves' disease, when thyrotoxicosis is controlled. This suggests that additional mechanisms not involving HLA-DR play a role in thyroid autoimmune disease.

One of the limitations of fine-needle aspiration (FNA) of the thyroid is difficulty in distinguishing the follicular variant (FV) of papillary thyroid carcinomas (PTC) from follicular neoplasms. By highlighting the "Orphan Annie-eyed" clear nuclei of the former, the Ultrafast Papanicolaou stain (UFP) easily separates these two entities. One thousand one hundred thirty-five ultrasound-guided FNAs of the thyroid were assessed by UFP with immediate biopsy results reported to the patients in a busy radiology office in Manhattan from November 1994 to December 1998. Of the 77 thyroid cancers resected, 22 were FVPTC and 17 were microcarcinomas (1 medullary carcinoma, 16 PTC). The rates of "unsatisfactory," "cancer," "suspicious for cancer," "follicular neoplasm," and "benign" cytology were 0.7%, 4.4%, 2.6%, 10.2%, and 82.1%, respectively and the cancer yields at surgery were 98%, 81.8%, 15.8%, and 0% respectively. Of the 1127 satisfactory FNAs in the series with a 2- to -6 years of clinical follow-up, a false-negative rate of 0% and a false-positive rate of 1.5% were obtained. Of the 169 surgical follow-ups with satisfactory FNAs, a sensitivity of 100%, specificity of 66.7%, positive predictive value of 87.4%, negative predictive value of 100%, and global accuracy of 89.9% were achieved. The paradoxical combination of low unsatisfactory rate and low false-negative rate is attributed to (1) the use of needle puncture without syringe to obtain enough microfollicles from the exceedingly bloody aspirates from follicular neoplasms for a diagnosis, (2) eliciting history of neck trauma to confirm hematomas, (3) using UFP to highlight the grape-like watery clear nuclei of FVPTC evident with a 4× objective, and (4) the precise guidance by ultrasound in sampling microcarcinomas.

We report the case of a patient with severe hypothyroid coma in whom hypothyroid-related neurogenic oropharyngeal dysphagia was suspected, videoscopically confirmed, and successfully treated. This complication has not previously been described, and may have contributed to the historically high mortality associated with severe cases of hypothyroid coma. In the future, the early detection and aggressive treatment of this complication and its sequelae should ensure a further reduction in mortality from hypothyroid coma.