scholarly journals A Change from Stimulatory to Blocking Antibody Activity in Graves’ Disease during Pregnancy1

1998 ◽  
Vol 83 (2) ◽  
pp. 514-518 ◽  
Author(s):  
A. W. C. Kung ◽  
B. M. Jones
Keyword(s):  
B Cells ◽  
Absolute Number ◽  
Human Thyroid ◽  
Graves Disease ◽  
Thyroid Cell ◽  
Antibody Activity ◽  
Inhibitory Antibody ◽  
Free T4 ◽  
Blocking Antibody ◽  
Blocking Antibodies

Remission of Graves’ disease (GD) during pregnancy with recrudescence after delivery is commonly observed. However, as pregnancy is associated with type 2 rather than type 1 cytokine production, a decrease in thyroid-stimulating antibody (TSAb) activity alone is unlikely to account for the remission during pregnancy. We hypothesized that a change in the antibody characteristics may occur as pregnancy advances. Fifteen women were studied in the first, second, and third trimesters of pregnancy and 4 months postpartum. TSH receptor antibodies were determined using human thyroid cell cultures, and lymphocyte subsets were measured by flow cytometry. Median TSAb (determined by cAMP release) decreased from 280% (96–3200) to 130% (range, 35–350; P < 0.05) during pregnancy, but no significant change was noted with the TSH binding inhibitory antibody (TBII; determined by RRA). Thyroid stimulation-blocking antibody (TSBAb; inhibition of TSH-stimulated cAMP release) increased from 16 ± 9% to 43 ± 16% (mean ± sd; P < 0.005). The increase in TSBAb was observed even among those patients who were in clinical remission before pregnancy. Overall, a negative correlation was observed between TSBAb activities and free T4 levels during pregnancy (r = −0.279; P < 0.05). Reciprocal changes in TSAb, TBII, and TSBAb levels were observed in the seven patients who relapsed during the postpartum period. In comparison, the healthy pregnant women (n = 14) were all negative for TSAb, TBII, and TSBAb throughout pregnancy. The absolute number of T lymphocytes, T helper cells, and natural killer cells, but not B cells, decreased significantly during pregnancy in both healthy women and GD patients. GD patients had significantly more CD5+ B cells at all stages of pregnancy compared to controls. In conclusion, a change in specificity from stimulatory to blocking antibodies was observed in GD patients during pregnancy and may contribute to the remission of GD during pregnancy.

Properties of human thyroidal and extrathyroidal TSH receptors

1981 ◽  
Vol 97 (4) ◽  
pp. 473-478 ◽  
Author(s):  
Krinos M. Trokoudes ◽  
Harold Michelsen ◽  
Arthur Kidd ◽  
Vas V. Row ◽  
Robert Volpé
Keyword(s):  
Thyroid Gland ◽  
Cell Membranes ◽  
Significant Inhibition ◽  
Membrane Receptors ◽  
Human Thyroid ◽  
Graves Disease ◽  
Thyroid Cell ◽  
Fat Cell ◽  
Acetylneuraminic Acid ◽  
Immunoglobulin Preparations

Abstract. The presence of high affinity receptor sites for thyroid-stimulating hormone (TSH) in the human is not limited to the thyroid gland. In this report, the properties of thyroidal and extrathyroidal TSH binding have been explored through the effect of various agents (TSI of Graves' disease, propranolol, hCG, N-acetylneuraminic acid, ACTH, insulin and L-thyroxine) on the receptors. Statistically significant inhibition of [125I]TSH binding occurred with all immunoglobulin preparations (TSI) of Graves' disease on thyroid cell membrane receptors. Thirty-three per cent of the same immunoglobulins showed significant inhibition of [125I]TSH binding when testicular cell membranes were used, while there was no such inhibition when renal cell membranes were utilized. D or L-propranolol at least doubled the [125I]TSH initial binding (Bo) to the thyroid membranes but had no effect on the testicular or fat cell binding. Human chorionic gonadotrophin (hCG) at concentrations of 400 USP units per 350 μl totally inhibited the [125I]TSH from binding to testicular membranes but not to thryroid or fat cell membranes. Conversely, the binding of [125I]hCG to human testicular membrane was inhibited by both stable hCG and TSH, but with human thyroid membrane, only TSH could inhibit binding of [125I]hCG. These data suggest that the TSH receptors in extrathyroidal tissues may not be identical to TSH receptors within the thyroid. TSH binding to thyroidal tissue was significantly suppressed by N-acetylneuraminic acid, was increased by D and L-propranolol and was unaffected by ACTH and insulin. L-thyroxine had a dose-related suppressive effect on the TSH binding, commencing at 50 μg/ml. While these effects were observed in vitro, it is possible that some of the above agents may also interfere with thyroid gland function in vivo. They suggest further that a number of interactions may take place at the TSH binding sites which could alter TSH binding and/or function.

Neuropilin-1 Receptor (NRP-1) Occupancy Induces Cell Death in Primary Chronic Lymphocytic Leukemia (CLL) B Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 586-586
Author(s):  
Grzegorz S. Nowakowski ◽  
Yean K. Lee ◽  
Neil Kay
Keyword(s):  
Flow Cytometry ◽  
Cell Death ◽  
B Cells ◽  
Gradient Centrifugation ◽  
Lymphocytic Leukemia ◽  
Vegf Receptor ◽  
Dependent Manner ◽  
Blocking Antibody ◽  
Induced Apoptosis ◽  
Blocking Antibodies

Abstract Background: We have previously shown that CLL B cells secrete vascular endothelial growth factor (VEGF) and express VEGF receptors: VEGFR-1 and VEGFR-2. Secreted VEGF protects CLL B cells from spontaneous and drug induced apoptosis via increased levels of Mcl-1 and XIAP; however, the exact mechanism of this process is unknown. In solid tumors, there is increasing evidence that signaling through VEGF receptor known as neuropillin-1 (NRP-1) is critical for VEGF induced resistance to apoptosis. Hypothesis: NRP-1 is expressed by CLL B cells and is critical for VEGF mediated protection from apoptosis. Methods: To demonstrate the presence of NRP-1 on CLL B cells, we conducted flow cytometry and immunoblotting. We then evaluated the ability of NRP-1 blocking antibodies to induce apoptosis of primary CLL B cells. To do this, circulating CLL B cells were isolated by density gradient centrifugation. Patient samples with greater than 80% of CD5+ CD19+ cells in a mononuclear cell population, as assessed by flow cytometry, were cultured in AIM-V media in 24-well plates at 1.5 x 106 cells/mL. To occupy NRP-1, we added NRP-1 blocking antibodies (Calbiochem, Darmstadt, Germany) at increasing concentrations (0.5 μg/mL -10 μg/mL) to cultured CLL B cells. Cell death was assessed using an annexin and propidium iodide flow assay after 24 h of in vitro culture. CLL cells cultured without antibodies and isotype nonspecific monoclonal antibodies were used as controls. Results: CLL B cells were found to express NRP-1 but not uniformly. Most patients had flow positive evidence for NRP-1 but a distinct percentage (35%) was very low (≤5%) or negative. However, immunoblot analysis revealed moderate to low levels of NRP-1 protein with evidence of tyrosine phosphorylation in all tested CLL patients (N=9). NRP-1 blocking antibodies, but not VEGF-R1 and -R2 blocking antibodies, induced apoptosis in a dose dependent manner in primary CLL cells (n=5, FIG.) at antibody concentrations starting at 1 μg/mL (p=0.003). The effect of the NRP-1 blockade varied between patients, with a median IC50, 1.5 μg/mL (range 0.5–2 μ/mL). Importantly, concentrations of 5 μg/mL and higher induced apoptosis in more than 90% of the CLL B cells. We also found that the sensitivity of CLL B cells to NRP-1 blocking antibody, in terms of apoptosis induction, was correlated with the number of NRP-1 receptors as assessed by flow cytometry. CLL B cell clones with no detectable NRP-1 had no induction of cell death when exposed to the NRP-1 blocking antibody. Finally, immunoprecipitation and immunoblot assays indicated that NRP-1 physically interacted with VEGF-R2 on CLL B cells. This suggests that NRP-1 could be enhancing VEGF binding affinity on VEGF-R2 to further increase the ability of VEGF to generate signals that lead to apoptosis modulation. Conclusion: We have found that NRP-1 blocking antibodies induce cell death in NRP-1 positive CLL B cells. Similar results using NRP-1 blocking peptides rather than blocking antibodies have been observed in breast cancer (Br J Cancer.2005 Jan 31;92(2):328–33). Our results suggest that NRP-1 represents an attractive therapeutic target in CLL and should be explored further. Figure Figure

scholarly journals Low-Dose Immunization with Adenovirus Expressing the Thyroid-Stimulating Hormone Receptor A-Subunit Deviates the Antibody Response toward That of Autoantibodies in Human Graves’ Disease

Endocrinology ◽  
2004 ◽  
Vol 145 (1) ◽  
pp. 228-233 ◽  
Author(s):  
Chun-Rong Chen ◽  
Pavel Pichurin ◽  
Gregorio D. Chazenbalk ◽  
Holly Aliesky ◽  
Yuji Nagayama ◽  
...  
Keyword(s):  
Antibody Response ◽  
Low Dose ◽  
Standard Dose ◽  
Thyroid Stimulating Hormone ◽  
Similar Proportion ◽  
Graves Disease ◽  
Antibody Activity ◽  
Blocking Antibody ◽  
A Subunit ◽  
Viral Dose

Abstract Immunization with adenovirus expressing the TSH receptor (TSHR) induces hyperthyroidism in 25–50% of mice. Even more effective is immunization with a TSHR A-subunit adenovirus (65–84% hyperthyroidism). Nevertheless, TSHR antibody characteristics in these mice do not mimic accurately those of autoantibodies in typical Graves’ patients, with a marked TSH-blocking antibody response. We hypothesized that this suboptimal antibody response was consequent to the standard dose of TSHR-adenovirus providing too great an immune stimulus. To test this hypothesis, we compared BALB/c mice immunized with the usual number (1011) and with far fewer viral particles (109 and 107). Regardless of viral dose, hyperthyroidism developed in a similar proportion (68–80%) of mice. We then examined the qualitative nature of TSHR antibodies in each group. Sera from all mice had TSH binding-inhibitory (TBI) activity after the second immunization, with TBI values in proportion to the viral dose. After the third injection, all groups had near-maximal TBI values. Remarkably, in confirmation of our hypothesis, immunization with progressively lower viral doses generated TSHR antibodies approaching the characteristics of autoantibodies in human Graves’ disease as follows: 1) lower TSHR antibody titers on ELISA and 2) lower TSH-blocking antibody activity without decrease in thyroid-stimulating antibody activity. In summary, low-dose immunization with adenovirus expressing the free TSHR A-subunit provides an induced animal model with a high prevalence of hyperthyroidism as well as TSHR antibodies more closely resembling autoantibodies in Graves’ disease.

Antibody-dependent cell-mediated growth inhibition of rat thyroid cells, FRTL-5, in patients with autoimmune thyroid diseases

1988 ◽  
Vol 118 (4) ◽  
pp. 580-586 ◽  
Author(s):  
Yasuhiro Iida ◽  
Kanji Kasagi ◽  
Yasutaka Tokuda ◽  
Keisuke Arai ◽  
Takashi Misaki ◽  
...  
Keyword(s):  
Growth Inhibition ◽  
Mononuclear Cell ◽  
Thymidine Incorporation ◽  
Normal Subjects ◽  
Human Thyroid ◽  
Graves Disease ◽  
Thyroid Cell ◽  
Chronic Thyroiditis ◽  
Thyroid Cells ◽  
Rat Thyroid

Abstract. We studied antibody-dependent mononuclear cell-mediated growth inhibition of thyroid cells in 18 untreated patients with Graves' disease, 18 patients with chronic thyroiditis, and 15 normal subjects by measuring the ability of their sera to inhibit [3H]thymidine incorporation into DNA in a rat thyroid cell line, FRTL-5, in the presence of normal mononuclear cells. [3H]thymidine incorporation was significantly inhibited in the presence of sera from patients with Graves' disease and chronic thyroiditis (P <0.001), whereas it was not affected in normal subjects. A significant correlation was observed between the inhibition of [3H]thymidine incorporation and the titre of anti-microsomal antibodies (P <0.05). The inhibitory effect on [3H]thymidine incorporation was significantly abolished when serum pre-absorbed with human thyroid membranes was used (P <0.005). These inhibitory effects on [3H]thymidine incorporation significantly correlated with those obtained by using IgG fractions (P <0.01). These data indicate that antibody-dependent mononuclear cell-mediated growth inhibtion may play a role in thyroid cell growth regulation in patients with autoimune thyroid disease.

scholarly journals Susceptibility Rather than Resistance to Hyperthyroidism Is Dominant in a Thyrotropin Receptor Adenovirus-Induced Animal Model of Graves’ Disease as Revealed by BALB/c-C57BL/6 Hybrid Mice

Endocrinology ◽  
2004 ◽  
Vol 145 (11) ◽  
pp. 4927-4933 ◽  
Author(s):  
Chun-Rong Chen ◽  
H. Aliesky ◽  
P. N. Pichurin ◽  
Y. Nagayama ◽  
S. M. McLachlan ◽  
...  
Keyword(s):  
Dominant Gene ◽  
F1 Hybrids ◽  
Graves Disease ◽  
Antibody Activity ◽  
Igg Subclass ◽  
Potential Candidate ◽  
T Helper ◽  
Blocking Antibody ◽  
Thyroid Stimulating Antibody ◽  
A Subunit

Abstract We investigated why TSH receptor (TSHR) adenovirus immunization induces hyperthyroidism more commonly in BALB/c than in C57BL/6 mice. Recent modifications of the adenovirus model suggested that using adenovirus expressing the TSHR A subunit (A-subunit-Ad), rather than the full-length TSHR, and injecting fewer viral particles would increase the frequency of hyperthyroidism in C57BL/6 mice. This hypothesis was not fulfilled; 65% of BALB/c but only 5% of C57BL/6 mice developed hyperthyroidism. TSH binding inhibitory antibody titers were similar in each strain. Functional TSHR antibody measurements provided a better indication for this strain difference. Whereas thyroid-stimulating antibody activity was higher in C57BL/6 than BALB/c mice, TSH blocking antibody activity was more potent in hyperthyroid-resistant C57BL/6 mice. F1 hybrids (BALB/c × C57BL/6) responded to A-subunit-Ad immunization with hyperthyroidism and TSHR antibody profiles similar to those of the hyperthyroid-susceptible parental BALB/c strain. In contrast, ELISA of TSHR antibodies revealed that the IgG subclass distribution in the F1 mice resembled the disease-resistant C57BL/6 parental strain. Because the IgG subclass distribution is dependent on the T helper 1/T helper 2 cytokine balance, this paradigm can likely be excluded as an explanation for susceptibility to hyperthyroidism. In summary, our data for BALB/c, C57BL/6, and F1 strains suggest that BALB/c mice carry a dominant gene(s) for susceptibility to induction of a thyroid-stimulating antibody/TSH blocking antibody balance that results in hyperthyroidism. Study of this genetic influence will provide useful information on potential candidate genes in human Graves’ disease.

scholarly journals Modulation of ACE-2 mRNA by inflammatory cytokines in human thyroid cells: a pilot study

Endocrine ◽  
2021 ◽  
Author(s):  
Francesca Coperchini ◽  
Gianluca Ricci ◽  
Laura Croce ◽  
Marco Denegri ◽  
Rubina Ruggiero ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
Combination Treatment ◽  
Primary Cultures ◽  
Human Thyroid ◽  
Thyroid Cell ◽  
Mrna Levels ◽  
Thyroid Cells ◽  
Tnf Α ◽  
Ifn Γ ◽  
Pro Inflammatory Cytokines

Abstract Introduction Angiotensin-converting-enzyme-2 (ACE-2) was demonstrated to be the receptor for cellular entry of SARS-CoV-2. ACE-2 mRNA was identified in several human tissues and recently also in thyroid cells in vitro. Purpose Aim of the present study was to investigate the effect of pro-inflammatory cytokines on the ACE-2 mRNA levels in human thyroid cells in primary cultures. Methods Primary thyroid cell cultures were treated with IFN-γ and TNF-α alone or in combination for 24 h. ACE-2 mRNA levels were measured by RT-PCR. As a control, the levels of IFN-γ inducible chemokine (CXCL10) were measured in the respective cell culture supernatants. Results The mean levels of ACE-2 mRNA increased after treatment with IFN-γ and TNF-α in all the thyroid cell preparations, while the combination treatment did not consistently synergically increase ACE-2-mRNA. At difference, CXCL10 was consistently increased by IFN-γ and synergically further increased by the combination treatment with IFN-γ + TNF-α, with respect to IFN-γ alone. Conclusions The results of the present study show that IFN-γ and, to a lesser extent TNF-α consistently increase ACE-2 mRNA levels in NHT primary cultures. More interestingly, the combined stimulation (proven to be effective according to the synergic effect registered for CXCL10) produces different responses in terms of ACE-2 mRNA modulation. These results would suggest that elevated levels of pro-inflammatory cytokines could facilitate the entering of the virus in cells by further increasing ACE-2 expression and/or account for the different degree of severity of SARS-COV-2 infection. This hypothesis deserves to be confirmed by further specific studies.

scholarly journals Trace Amine-Associated Receptor 1 Trafficking to Cilia of Thyroid Epithelial Cells

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1518
Author(s):  
Maria Qatato ◽  
Vaishnavi Venugopalan ◽  
Alaa Al-Hashimi ◽  
Maren Rehders ◽  
Aaron D. Valentine ◽  
...  
Keyword(s):  
Cell Surface ◽  
Cell Lines ◽  
Human Thyroid ◽  
Apical Plasma Membrane ◽  
Thyroid Cell ◽  
Thyroid Cells ◽  
Trace Amine ◽  
Rat Thyroid

Trace amine-associated receptor 1 (rodent Taar1/human TAAR1) is a G protein-coupled receptor that is mainly recognized for its functions in neuromodulation. Previous in vitro studies suggested that Taar1 may signal from intracellular compartments. However, we have shown Taar1 to localize apically and on ciliary extensions in rodent thyrocytes, suggesting that at least in the thyroid, Taar1 may signal from the cilia at the apical plasma membrane domain of thyrocytes in situ, where it is exposed to the content of the follicle lumen containing putative Taar1 ligands. This study was designed to explore mouse Taar1 (mTaar1) trafficking, heterologously expressed in human and rat thyroid cell lines in order to establish an in vitro system in which Taar1 signaling from the cell surface can be studied in future. The results showed that chimeric mTaar1-EGFP traffics to the apical cell surface and localizes particularly to spherical structures of polarized thyroid cells, procilia, and primary cilia upon serum-starvation. Moreover, mTaar1-EGFP appears to form high molecular mass forms, possibly homodimers and tetramers, in stably expressing human thyroid cell lines. However, only monomeric mTaar1-EGFP was cell surface biotinylated in polarized human thyrocytes. In polarized rat thyrocytes, mTaar1-EGFP is retained in the endoplasmic reticulum, while cilia were reached by mTaar1-EGFP transiently co-expressed in combination with an HA-tagged construct of the related mTaar5. We conclude that Taar1 trafficking to cilia depends on their integrity. The results further suggest that an in vitro cell model was established that recapitulates Taar1 trafficking in thyrocytes in situ, in principle, and will enable studying Taar1 signaling in future, thus extending our general understanding of its potential significance for thyroid autoregulation.

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