scholarly journals Microarray Analysis of Cytokine Activation of Apoptosis Pathways in the Thyroid

Endocrinology ◽  
2007 ◽  
Vol 148 (10) ◽  
pp. 4844-4852 ◽  
Author(s):  
Su He Wang ◽  
Mary Van Antwerp ◽  
Rork Kuick ◽  
Paul G. Gauger ◽  
Gerard M. Doherty ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
P38 Mapk ◽  
Human Thyroid ◽  
Autoimmune Thyroid Diseases ◽  
Thyroid Cells ◽  
Mapk Phosphorylation ◽  
Autoimmune Thyroid ◽  
Cytokine Activation ◽  
Ifn Γ ◽  
Sb 203580

It has been suggested that Fas-mediated apoptosis plays an important role in the pathogenesis of autoimmune thyroid diseases. Our previous studies have demonstrated that normal primary thyroid epithelial cells are resistant to Fas-mediated apoptosis, but the resistance can be overcome by pretreatment with a combination of interferon-γ (IFN-γ) and IL-1β. To understand the molecular mechanism responsible for the IFN-γ/IL-1β effects, we profiled changes in the transcription induced by these two cytokines in normal human thyroid cells, using cDNA microarrays. We found that IFN-γ/IL-1β showed a significant increase in apoptosis-related genes such as inducible nitric oxide synthase (iNOS), receptor-interacting protein 2 (RIP2), and caspases 10. These increases were confirmed by other methods, including real-time PCR and Western blot. Furthermore, the sensitization of primary thyroid epithelial cells to Fas-mediated apoptosis by IFN-γ/IL-1β was significantly blocked by a general caspase inhibitor, z-VAD, or by the combination of two specific individual caspase inhibitors. In addition, our results showed that IFN-γ/IL-1β enhance p38 MAPK phosphorylation and that SB 203580, a p38 MAPK inhibitor, can inhibit IFN-γ/IL-1β-induced p38 MAPK phosphorylation. SB 203580 also significantly prevented cytokine-induced iNOS expression and caspase activation and thus blocked Fas-mediated apoptosis of thyroid cells sensitized by IFN-γ/IL-1β. In conclusion, our data suggest that both p38 MAPK and iNOS are involved in IFN-γ/IL-1β-induced sensitization of the thyroid cells to Fas-mediated apoptosis via the activation of caspases 3, 7, and 10 and that this pathway may be further activated by BID. This hints that inflammatory cytokines regulate death-receptor-mediated apoptosis at multiple points in the process.

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.

p38 mitogen-activated protein kinase inhibits calcium-dependent chloride secretion in T84 colonic epithelial cells

2003 ◽  
Vol 284 (2) ◽  
pp. C339-C348 ◽  
Author(s):  
Stephen J. Keely ◽  
Kim E. Barrett
Keyword(s):  
Protein Kinase ◽  
Epithelial Cells ◽  
Signaling Pathway ◽  
P38 Mapk ◽  
Mitogen Activated Protein Kinase ◽  
Muscarinic Agonist ◽  
Colonic Epithelial Cells ◽  
Mitogen Activated Protein ◽  
Intracellular Ca ◽  
Sb 203580

We have previously shown that Ca2+-dependent Cl−secretion across intestinal epithelial cells is limited by a signaling pathway involving transactivation of the epidermal growth factor receptor (EGFR) and activation of ERK mitogen-activated protein kinase (MAPK). Here, we have investigated a possible role for p38 MAPK in regulation of Ca2+-dependent Cl− secretion. Western blot analysis of T84 colonic epithelial cells revealed that the muscarinic agonist carbachol (CCh; 100 μM) stimulated phosphorylation and activation of p38 MAPK. The p38 inhibitor SB-203580 (10 μM) potentiated and prolonged short-circuit current ( I sc) responses to CCh across voltage-clamped T84 cells to 157.4 ± 6.9% of those in control cells ( n = 21; P < 0.001). CCh-induced p38 phosphorylation was attenuated by the EGFR inhibitor tyrphostin AG-1478 (0.1 nM–10 μM) and by the Src family kinase inhibitor PP2 (20 nM–2 μM). The effects of CCh on p38 phosphorylation were mimicked by thapsigargin (TG; 2 μM), which specifically elevates intracellular Ca2+, and were abolished by the Ca2+ chelator BAPTA-AM (20 μM), implying a role for intracellular Ca2+ in mediating p38 activation. SB-203580 (10 μM) potentiated I sc responses to TG to 172.4 ± 18.1% of those in control cells ( n= 18; P < 0.001). When cells were pretreated with SB-203580 and PD-98059 to simultaneously inhibit p38 and ERK MAPKs, respectively, I sc responses to TG and CCh were significantly greater than those observed with either inhibitor alone. We conclude that Ca2+-dependent agonists stimulate p38 MAPK in T84 cells by a mechanism involving intracellular Ca2+, Src family kinases, and the EGFR. CCh-stimulated p38 activation constitutes a similar, but distinct and complementary, antisecretory signaling pathway to that of ERK MAPK.

scholarly journals Antithyroperoxidase Antibody-Dependent Cytotoxicity in Autoimmune Thyroid Disease

2008 ◽  
Vol 93 (3) ◽  
pp. 929-934 ◽  
Author(s):  
Sandra A. Rebuffat ◽  
Brigitte Nguyen ◽  
Bruno Robert ◽  
Françoise Castex ◽  
Sylvie Peraldi-Roux
Keyword(s):  
Cell Line ◽  
Thyroid Disease ◽  
Autoimmune Thyroid Disease ◽  
Human Monocyte ◽  
Human Thyroid ◽  
Chromium Release Assay ◽  
Thyroid Cells ◽  
Effector Cells ◽  
Autoimmune Thyroid ◽  
Chromium Release

Abstract Context: Thyroid antibody-dependent cytotoxicity has been reported in autoimmune thyroid disease (AITD). Indeed, the role of thyroperoxidase (TPO) autoantibodies (aAbs) in complement-mediated damage by binding to TPO expressed on the surface of human thyroid cells was demonstrated, whereas their activity in antibody-dependent cell cytotoxicity (ADCC) is not well established. Objective: The aim of this study was to define the partners involved in antibody and complement-dependent cytotoxicity (CDC) in AITD and characterize which effector cells are involved in cytotoxicity mediated by anti-TPO aAbs using a chromium release assay. Results: The relative capability of anti-TPO aAbs to mediate ADCC using human thyroid cells in culture varies from 11 to 74.5%, depending on the effectors cells used. The human monocyte cell line HL60 gives a better lysis than the THP-1 cell line as effector cells. It seems obvious that the mechanism of ADCC is mediated quite exclusively by FcγRI. Indeed, the two effector cell lines differ by the level of the FcγRI expression (91.83% for HL-60 cells and 22.55%t for the THP-1). In addition to ADCC, the anti-TPO aAbs mediate the destruction of thyrocytes by CDC (56%). Conclusions: These results demonstrate that anti-TPO aAbs can damage cultured thyroid cells by ADCC and CDC mechanisms. The monocytes, via their FcγRI, are important effector cells in ADCC mediated by anti-TPO aAbs and may contribute with T cells to the destruction of thyroid gland in AITD.

Conditional immortalization of human thyroid epithelial cells: a tool for analysis of oncogene action

1990 ◽  
Vol 10 (10) ◽  
pp. 5365-5377
Author(s):  
D Wynford-Thomas ◽  
J A Bond ◽  
F S Wyllie ◽  
J S Burns ◽  
E D Williams ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Simian Virus 40 ◽  
Cell Types ◽  
Human Thyroid ◽  
Restrictive Temperature ◽  
Permissive Temperature ◽  
Thyroid Cells ◽  
Early Region ◽  
Sv40 Early Region

To overcome the difficulty of assessing oncogene action in human epithelial cell types, such as thyroid, which have limited proliferative potential in culture, we have explored the use of temperature-sensitive (ts) mutants of simian virus 40 (SV40) early region to create conditionally immortalized epithelial cell lines. Normal primary cultures of human thyroid follicular cells were transfected with a plasmid containing the SV40 early region from mutant tsA58. Expanding epithelial colonies were observed after 2 to 3 months, all of which grew to greater than 200 population doublings without crisis. All showed tight temperature dependence for growth. After switch-up to the restrictive temperature (40.5 degrees C), no further increase in cell number was seen after 1 to 2 days. However, DNA synthesis declined much more slowly; the dissociation from cell division led to marked polyploidy. Viability was maintained for up to 2 weeks. Introduction of an inducible mutant ras gene into ts thyroid cells led, as expected, to morphological transformation at the permissive temperature when ras was induced. Interestingly, this was associated with a marked reduction in net growth rate. At the restrictive temperature, induction of mutant ras caused rapid cell death. These results demonstrate the utility of a ts SV40 mutant to permit the study of oncogene action in an otherwise nonproliferative target cell and reveal important differences in the interaction between ras and SV40 T in these epithelial cells compared with previously studied cell types.

Historical Review

2020 ◽  
pp. 1-4
Keyword(s):  
Needle Biopsy ◽  
Historical Review ◽  
Hashimoto Thyroiditis ◽  
Thyroid Disorders ◽  
Historical Overview ◽  
Autoimmune Thyroid Diseases ◽  
Thyroid Cells ◽  
Autoimmune Thyroid ◽  
Japanese Surgeon ◽  
Over Time

Hashimoto thyroiditis (HT) is part of the spectrum of autoimmune thyroid diseases characterized by the destruction of thyroid cells by various cell- and antibody-mediated immune processes. It was first described by the Japanese surgeon Hakaru Hashimoto (1981-1934). It was not until 1956 when a link between antibodies to thyroid cells present in the serum of patients and HT was made. Over time, our understanding of the immunologic pathways involved in HT has evolved. We now recognize the association of this disease with other autoimmune diseases and thyroid cancer. The increasing use of the needle biopsy and serologic tests for antibodies have led to much more frequent recognition, and there is reason to believe that it may be increasing in frequency. It is now one of the most common thyroid disorders. This chapter gives a historical overview of Hashimoto's disease.

Antibodies to membrane antigens in autoimmune thyroid disease

1987 ◽  
Vol 116 (1) ◽  
pp. 13-20 ◽  
Author(s):  
J. Furmaniak ◽  
J. Bradbury ◽  
B. Rees Smith
Keyword(s):  
Solid Phase ◽  
Protein A ◽  
Human Thyroid ◽  
Intermediate Filament Protein ◽  
Autoimmune Thyroid Diseases ◽  
Autoimmune Thyroid ◽  
Microsomal Antibody ◽  
Membrane Antigens ◽  
Solid Phase Assay ◽  
Microsomal Antigen

Abstract. The possibility that sera from patients with autoimmune thyroid diseases contain autoantibodies to thyroid membrane proteins distinct from microsomal antigen and the TSH receptor has been investigated using (a) solid phase assay system based on human thyroid membranes and 125I-labelled protein A and (b) immunoprecipitation of detergent solubilized 125I-labelled thyroid membranes followed by gel electrophoresis and autoradiography. In the solid phase assay binding to membranes showed a highly significant correlation with binding to microsomes (r = 0.82; P < 0.001; N = 82) indicating that the interaction between the serum and the membranes was due principally to microsomal antibody binding to microsomal antigen contaminating the membrane preparations. However, there were some discrepancies suggesting that an additional antigen-antibody system was involved. This possibility was then investigated using immunoprecipitation of 125I-labelled thyroid membranes. A labelled protein with mol wt 54 K was specifically immunoprecipitated (relative to normal pool serum) by 3 out of 4 sera from patients with Graves' disease who showed high binding to thyroid membranes. A further 4 sera from such patients with low membrane binding affinity failed to immunoprecipitate the 54 K protein. Sera from some patients with Hashimoto's disease and some patients with rheumatoid arthritis and one patient with Addison's disease also immunoprecipitated the 54 K protein from solubilized thyroid membranes. These studies suggested that antibodies interacting with the 54 K protein contributed to the discrepancies between thyroid membrane and microsome binding. However, the 54 K protein was also immunoprecipitated from detergent solubilized membranes prepared from human placenta, skeletal muscle and adrenal tissue. Immunoprecipitation studies with antisera to cytoskeleton proteins suggested that the 54 K band was the intermediate filament protein desmin. Consequently, thyroid specific antibody-antigen systems distinct from those involving microsomal antibody (or thyroglobulin antibody) could not be detected in thyroid membranes by immunoprecipitation.

A role for insulin-like growth factor-I in the regulation of human thyroid cell growth by thyrotrophin

1989 ◽  
Vol 123 (3) ◽  
pp. 495-NP ◽  
Author(s):  
C. A. Ollis ◽  
D. J. Hill ◽  
D. S. Munro
Keyword(s):  
Monoclonal Antibody ◽  
Growth Factor ◽  
Epithelial Cells ◽  
Human Thyroid ◽  
Immunocytochemical Staining ◽  
Thymidine Uptake ◽  
Thyroid Cells ◽  
Factor I ◽  
Igf I ◽  
Growth Factor I

ABSTRACT Human thyroid epithelial cells in monolayer culture were found to release radioimmunoassayable insulinlike growth factor-I (IGF-I) over a 48-h culture period in serum-free medium. In the presence of supraphysiological concentrations of TSH (1–100 mU/ml) known to be inhibitory to DNA synthesis by human thyroid cells, the release of IGF-I was found to be inhibited in six thyroid cultures studied. In only one out of the six was IGF-I release increased in the presence of physiological mitogenic concentrations of TSH (0·1–100 μU/ml). Human thyroid fibroblasts, established by long-term culture of thyroid epithelial cells under fibroblast-selective conditions, also secreted IGF-I which was unaffected by the presence of TSH at both low and high concentrations. Using a monoclonal antibody against human IGF-I, monolayer cultures of both human thyroid epithelial cells and human thyroid fibroblasts showed positive immunocytochemical staining for IGF peptide. However, fixed sections of intact thyroid tissue only showed positive staining for IGF peptide associated with the fibrous layers surrounding the thyroid follicle, with no staining of the follicular epithelial cells. The growth of human thyroid epithelial cells was also found to be increased by IGF-I (25–100 ng/ml) added in medium plus 1 % fetal calf serum as assessed by the incorporation of [3H]thymidine into DNA. In the presence of a monoclonal antibody to IGF-I the increase in [3H]thymidine uptake in response to IGF-I was abolished as was that seen in response to TSH. This study indicates a possible paracrine/autocrine role of IGF-I in the regulation of human thyroid epithelial cell proliferation by interaction with TSH. Journal of Endocrinology (1989) 123, 495–500

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