Name: generation of a mouse spontaneous autoimmune thyroiditis model
Uploaded: 2023-03-17T14:05:00
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Mouse monoclonal antibodies to human TPO (used as positive controls) were provided by Dr. P. Carayon and Dr. J. Ruf (Marseille, France). The authors thank all the participants in this study and the members of our research team. This work was in part supported by grants from the Postdoctoral Sustentation Fund of West China Hospital, Sichuan University, China (2020HXBH057) and the Sichuan Province Science and Technology Support Program (Project No. 2021YFS0166)

The protocol described below was approved by the care and use guidelines established by the Institutional Animal Care and Use Committee of Sichuan University.
1. Preparation
<ol>
	<li>House all mice in specific pathogen-free conditions under 12 h light-dark cycles (beginning at 07:00 a.m. and 07:00 p.m., respectively). Maintain the room temperature at 22 &#176;C. Change the bedding materials every week. Provide adequate quantities of standard rodent chow and water.</li>
	<li...

<ol>
	<li>Ralli, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hashimoto's+thyroiditis:+An+update+on+pathogenic+mechanisms,+diagnostic+protocols,+therapeutic+strategies,+and+potential+malignant+transformation.">Hashimoto's thyroiditis: An update on pathogenic mechanisms, diagnostic protocols, therapeutic strategies, and potential malignant transformation.</a> Autoimmunity Reviews. 19 (10), 102649 (2020).</li><li>Zhang, Q. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymphocyte+infiltration+and+thyrocyte+destruction+are+driven+by+stromal+and+immune+cell+components+in+Hashimoto's+thyroiditis.">Lymphocyte infiltration and thyrocyte destruction are driven by stromal and immune cell components in Hashimoto's thyroiditis.</a> Nature Communications. 13 (1), 775 (2022).</li><li>Ruggeri, R. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoimmune+comorbidities+in+Hashimoto's+thyroiditis:+different+patterns+of+association+in+adulthood+and+childhood/adolescence.">Autoimmune comorbidities in Hashimoto's thyroiditis: different patterns of association in adulthood and childhood/adolescence.</a> European Journal of Endocrinology. 176 (2), 133-141 (2017).</li><li>Resende de Paiva, C., Gr&#248;nh&#248;j, C., Feldt-Rasmussen, U., von Buchwald, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+between+Hashimoto's+thyroiditis+and+thyroid+cancer+in+64,628+patients.">Association between Hashimoto's thyroiditis and thyroid cancer in 64,628 patients.</a> Frontiers in Oncology. 7, 53 (2017).</li><li>Ehlers, M., Schott, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hashimoto's+thyroiditis+and+papillary+thyroid+cancer:+are+they+immunologically+linked.">Hashimoto's thyroiditis and papillary thyroid cancer: are they immunologically linked.</a> Trends in Endocrinology and Metabolism. 25 (12), 656-664 (2014).</li><li>Medenica, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+cell+and+gene+therapies+in+the+treatment+of+infertility+in+patients+with+thyroid+autoimmunity.">The role of cell and gene therapies in the treatment of infertility in patients with thyroid autoimmunity.</a> International Journal of Endocrinology. 2022, 4842316 (2022).</li><li>Rose, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+autoimmune+thyroiditis:+the+first+decade.">The genetics of autoimmune thyroiditis: the first decade.</a> Journal of Autoimmunity. 37 (2), 88-94 (2011).</li><li>Kolypetri, P., King, J., Larijani, M., Carayanniotis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genes+and+environment+as+predisposing+factors+in+autoimmunity:+acceleration+of+spontaneous+thyroiditis+by+dietary+iodide+in+NOD.H2(h4)+mice.">Genes and environment as predisposing factors in autoimmunity: acceleration of spontaneous thyroiditis by dietary iodide in NOD.H2(h4) mice.</a> International Reviews of Immunology. 34 (6), 542-556 (2015).</li><li>Terplan, K. L., Witebsky, E., Rose, N. R., Paine, J. R., Egan, R. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+mechanisms+of+GABA+receptor+autoimmune+encephalitis.">Structural mechanisms of GABA receptor autoimmune encephalitis.</a> Cell. 185 (14), 2469-2477 (2022).</li><li>Pudifin, D. J., Duursma, J., Brain, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+autoimmune+thyroiditis+in+the+vervet+monkey.">Experimental autoimmune thyroiditis in the vervet monkey.</a> Clinical and Experimental Immunology. 29 (2), 256-260 (1977).</li><li>Esquivel, P. S., Rose, N. R., Kong, Y. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+autoimmunity+in+good+and+poor+responder+mice+with+mouse+thyroglobulin+and+lipopolysaccharide.">Induction of autoimmunity in good and poor responder mice with mouse thyroglobulin and lipopolysaccharide.</a> The Journal of Experimental Medicine. 145 (5), 1250-1263 (1977).</li><li>Kong, Y. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLA-DRB1+polymorphism+determines+susceptibility+to+autoimmune+thyroiditis+in+transgenic+mice:+definitive+association+with+HLA-DRB1*0301+(DR3)+gene.">HLA-DRB1 polymorphism determines susceptibility to autoimmune thyroiditis in transgenic mice: definitive association with HLA-DRB1*0301 (DR3) gene.</a> The Journal of Experimental Medicine. 184 (3), 1167-1172 (1996).</li><li>Kotani, T., Umeki, K., Hirai, K., Ohtakia, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+murine+thyroiditis+induced+by+porcine+thyroid+peroxidase+and+its+transfer+by+the+antigen-specific+T+cell+line.">Experimental murine thyroiditis induced by porcine thyroid peroxidase and its transfer by the antigen-specific T cell line.</a> Clinical and Experimental Immunology. 80 (1), 11-18 (1990).</li><li>Ng, H. P., Banga, J. P., Kung, A. W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+murine+model+of+autoimmune+thyroiditis+induced+with+homologous+mouse+thyroid+peroxidase.">Development of a murine model of autoimmune thyroiditis induced with homologous mouse thyroid peroxidase.</a> Endocrinology. 145 (2), 809-816 (2004).</li><li>Ng, H. P., Kung, A. W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+autoimmune+thyroiditis+and+hypothyroidism+by+immunization+of+immunoactive+T+cell+epitope+of+thyroid+peroxidase.">Induction of autoimmune thyroiditis and hypothyroidism by immunization of immunoactive T cell epitope of thyroid peroxidase.</a> Endocrinology. 147 (6), 3085-3092 (2006).</li><li>Ellis, J. S., Braley-Mullen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+by+which+B+cells+and+regulatory+T+Cells+influence+development+of+murine+organ-specific+autoimmune+diseases.">Mechanisms by which B cells and regulatory T Cells influence development of murine organ-specific autoimmune diseases.</a> Journal of Clinical Medicine. 6 (2), 13 (2017).</li><li>Fang, Y., Yu, S., Braley-Mullen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contrasting+roles+of+IFN-gamma+in+murine+models+of+autoimmune+thyroid+diseases.">Contrasting roles of IFN-gamma in murine models of autoimmune thyroid diseases.</a> Thyroid. 17 (10), 989-994 (2007).</li><li>Fang, Y., Zhao, L., Yan, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemokines+as+novel+therapeutic+targets+in+autoimmune+thyroiditis.">Chemokines as novel therapeutic targets in autoimmune thyroiditis.</a> Recent Patents on DNA &amp; Gene Sequences. 4 (1), 52-57 (2010).</li><li>Chen, C. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibodies+to+thyroid+peroxidase+arise+spontaneously+with+age+in+NOD.H-2h4+mice+and+appear+after+thyroglobulin+antibodies.">Antibodies to thyroid peroxidase arise spontaneously with age in NOD.H-2h4 mice and appear after thyroglobulin antibodies.</a> Endocrinology. 151 (9), 4583-4593 (2010).</li><li>Ruf, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+immunological+structure+and+biochemical+properties+of+human+thyroid+peroxidase.">Relationship between immunological structure and biochemical properties of human thyroid peroxidase.</a> Endocrinology. 125 (3), 1211-1218 (1989).</li><li>McLachlan, S. M., Aliesky, H. A., Chen, C. R., Chong, G., Rapoport, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breaking+tolerance+in+transgenic+mice+expressing+the+human+TSH+receptor+A-subunit:+thyroiditis,+epitope+spreading+and+adjuvant+as+a+'double+edged+sword'.">Breaking tolerance in transgenic mice expressing the human TSH receptor A-subunit: thyroiditis, epitope spreading and adjuvant as a 'double edged sword'.</a> PLoS One. 7 (9), e43517 (2012).</li><li>McLachlan, S. M., Aliesky, H. A., Chen, C. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breaking+tolerance+in+transgenic+mice+expressing+the+human+TSH+receptor+A-subunit:+thyroiditis,+epitope+spreading+and+adjuvant+as+a+'double+edged+sword’[J].">Breaking tolerance in transgenic mice expressing the human TSH receptor A-subunit: thyroiditis, epitope spreading and adjuvant as a 'double edged sword’[J].</a> PLoS One. 7 (9), e43517 (2012).</li><li>Hutchings, P. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Both+CD4(+)+T+cells+and+CD8(+)+T+cells+are+required+for+iodine+accelerated+thyroiditis+in+NOD+mice.">Both CD4(+) T cells and CD8(+) T cells are required for iodine accelerated thyroiditis in NOD mice.</a> Cellular Immunology. 192 (2), 113-121 (1999).</li><li>Xue, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+changes+of+CD4+CD25+++regulatory+T+cells+in+NOD.H-2h4+mice+with+iodine-induced+autoimmune+thyroiditis.">Dynamic changes of CD4+CD25 + regulatory T cells in NOD.H-2h4 mice with iodine-induced autoimmune thyroiditis.</a> Biological Trace Element Research. 143 (1), 292-301 (2011).</li><li>Hou, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+halofuginone+on+the+pathogenesis+of+autoimmune+thyroid+disease+in+different+mice+models.">Effect of halofuginone on the pathogenesis of autoimmune thyroid disease in different mice models.</a> Endocrine, Metabolic &amp; Immune Disorders Drug Targets. 17 (2), 141-148 (2017).</li><li>McLachlan, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissociation+between+iodide-induced+thyroiditis+and+antibody-mediated+hyperthyroidism+in+NOD.H-2h4+mice.">Dissociation between iodide-induced thyroiditis and antibody-mediated hyperthyroidism in NOD.H-2h4 mice.</a> Endocrinology. 146 (1), 294-300 (2005).</li><li>Danailova, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nutritional+management+of+thyroiditis+of+hashimoto.">Nutritional management of thyroiditis of hashimoto.</a> International Journal of Molecular Sciences. 23 (9), 5144 (2022).</li><li>Carayanniotis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+parameters+linking+thyroglobulin+iodination+with+autoimmune+thyroiditis.">Molecular parameters linking thyroglobulin iodination with autoimmune thyroiditis.</a> Hormones. 10 (1), 27-35 (2011).</li><li>Verginis, P., Li, H. S., Carayanniotis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tolerogenic+semimature+dendritic+cells+suppress+experimental+autoimmune+thyroiditis+by+activation+of+thyroglobulin-specific+CD4+CD25++T+cells.">Tolerogenic semimature dendritic cells suppress experimental autoimmune thyroiditis by activation of thyroglobulin-specific CD4+CD25+ T cells.</a> Journal of Immunology. 174 (11), 7433-7439 (2005).</li><li>Flynn, J. 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HT occurs due to an autoimmune system disorder caused by lymphocytes infiltrating the thyroid gland, further impairing thyroid function, while producing thyroid-specific antibodies. Serum TSH, TgAb, and TPOAb levels in HT patients are significantly elevated27. At present, two main kinds of murine models are widely used to study the etiology of autoimmune thyroiditis: EAT and SAT29. EAT mice are mostly immunized using proteins and adjuvants to create an abnormal immune envir...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/64609">Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model</a>. The Protocol section was updated.
Step 3.1.1 of the Protocol was updated from:
After the induction, anesthetize the mice with a volume of 0.01 mL/g anesthetic by intraperitoneal injection. Prepare the anesthetic by mixing midazolam (40 &#181;g/100 &#181;L for sedation), medetomidine (7.5 &#181;g/100 &#181;L for sedation), and butorphanol tartrate (50 &#181;g/100 &#181;L for analgesia) in phosphate-buffered saline (PBS).
to
After the induction, anesthetize the mice with a volume of 0.01 mL/g anesthetic by intraperitoneal injection. Prepare the anesthetic by mixing midazolam (40 &#181;g/100 &#181;L for sedation), medetomidine (7.5 &#181;g/100 &#181;L for sedation), and butorphanol tartrate (50 &#181;g/100 &#181;L for analgesia) in phosphate-buffered saline (PBS). 
NOTE: The specific concentrations of each component in the anesthesia mixture are: midazolam 13.33&#181;g/100&#181;L, medetomidine 2.5&#181;g/100&#181;L, and butorphanol 16.7&#181;g/100&#181;L. For specific dosages used in mice, the doses are: midazolam 4&#181;g/g, medetomidine 0.75&#181;g/g, and butorphanol 1.67&#181;g/g.&#160;Anesthesia depth was confirmed when the mouse&#39;s limb muscles relaxed, the whiskers had no touch response, and there was loss of&#160; pedal reflex.
Step 3.1.2 of the Protocol was updated from:
After the mice are anesthetized, cut off their whiskers with ophthalmic scissors to prevent blood from flowing down the whiskers and causing hemolysis. Fix the mouse with one hand and press the skin of the eye to make the eyeball protrude. Quickly remove the eyeball and draw 1 mL of blood into the microcentrifuge tube via a capillary tube.
to
After the mice are anesthetized, prepare the peripheral blood samples, by fixing the mouse with one hand and pressing the eye skin to protrude the eyeball. Then, insert the capillary tube into the inner corner of the eye and penetrate at a 30-45 degree angle to the plane of the nostril. Apply pressure while gently rotating the capillary tube. Blood will flow into the tube via capillary action.
Step 3.2.1 of the Protocol was updated from:
Dissect the chest wall to expose the heart, cut open the right atrium, and infuse saline into the left ventricle by an intravenous infusion needle attached to a 20 mL syringe until the tissue turns white.
to
Humanely euthanize the animal according to the institutional policies. Then, dissect the chest wall to expose the heart, cut open the right atrium, and infuse saline into the left ventricle by an intravenous infusion needle attached to a 20 mL syringe until the tissue turns white.

An erratum was issued for:&#160;<a href="https://www.jove.com/t/64609">Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model</a>. The Protocol section was updated.

Erratum: Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model

Hashimoto&#39;s thyroiditis (HT), also known as chronic lymphocytic thyroiditis or autoimmune thyroiditis, was first reported in 19121. HT is characterized by lymphocyte infiltration and damage to thyroid follicular tissue. Laboratory tests are mainly manifested as increasing thyroid-specific antibodies, including anti-thyroglobulin antibody (TgAb) and anti-thyroid peroxidase antibody (TPOAb)2. The incidence of HT is in the range of 0.4%-1.5%, accounting for 20%-25% of all thyroid diseases, and this value has increased in recent years3. In addition, a large number of studies have reported that HT is associated with the oncogenesis and recurrence of papillary thyroid carcinoma (PTC)4,5; the potential mechanisms are still controversial. Autoimmune thyroiditis is also an important factor in female infertility6. Therefore, the pathogenesis of HT needs to be clear, for which a stable and simple animal model is essential.
To study the etiology of HT, two main kinds of murine models have been employed, including experimental autoimmune thyroiditis (EAT) and spontaneous autoimmune thyroiditis (SAT) in the present studies7,8. Susceptible mice were immunized with specific thyroid antigens (including the crude thyroid, purified thyroglobulin [TG], thyroid peroxidase [TPO], recombinant TPO ectodomain, and selected TPO peptides) to establish the EAT murine model. In addition, the adjuvants, including lipopolysaccharide (LPS), complete Freund&#39;s adjuvant (CFA), and other unusual adjuvants, are also used during the immunization to break down immune tolerance9,10,11,12,13,14,15,16,17.
The SAT model is an important model to study the spontaneous development of autoimmune thyroiditis, which is based on NOD.H-2h4 mice. The NOD.H-2h4 mouse is a new strain obtained from the cross of NOD and B10.A(4R) mice, followed by multiple backcrosses to NOD, with the autoimmune thyroiditis susceptibility gene IAk18,19. NOD.H-2h4 mice do not develop diabetes, but have a high incidence of autoimmune thyroiditis and Sjogren&#39;s syndrome (SS)19. Studies have found that intracellular adhesion molecule-1 (ICAM-1) is highly expressed in the thyroid tissue of NOD.H-2h4 mice at 3-4 weeks of age. Moreover, with the increase in iodine intake, the immunogenicity of the thyroglobulin molecule is enhanced, which further upregulates the expression of ICAM-1, which plays an important role in the process of monocyte infiltration21. This model simulates the autoimmune process while verifying the relationship between iodine dose and disease severity. The established method is stable, with a high probability of success. The SAT model has been applied to induce autoimmune thyroiditis for many years and continues to be an effective method to study the pathogenesis of autoimmune thyroiditis. However, the current construction method of the EAT model is more complicated and expensive; different laboratories use different immunization methods and injection sites. Furthermore, mice with different genetic backgrounds have different rates of induction, which need further study to reveal the potent mechanism.
However, the development of thyroiditis in the SAT model is associated with sodium iodide, sexual dimorphism, and the rearing conditions. To reveal the appropriate procedure of autoimmune thyroiditis in the SAT model, this article described the method of induction of autoimmune thyroiditis in different conditions. In addition, it allows for the study of the pathogenesis and immunological progress of autoimmune thyroiditis in different stages of this disease.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Butorphanol tartrate</td>
			<td>Supelco</td>
			<td>L-044&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Dexmedetomidine hydrochloride&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>145108-58-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Enzyme-linked immunosorbent assay (ELISA) well</td>
			<td>Sigma-Aldrich</td>
			<td>M9410-1CS</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>macklin</td>
			<td>64-17-5&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Freund&#8217;s Adjuvant, Complete&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>F5881&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Freund&#8217;s Adjuvant, Incomplete&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>F5506</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-Mouse IgG&#160;</td>
			<td>invitrogen</td>
			<td>SA5-10275&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Midazolam solution&#160;</td>
			<td>Supelco</td>
			<td>M-908&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse/rat thyroxine (T4) ELISA</td>
			<td>Calbiotech</td>
			<td>DKO045</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>macklin</td>
			<td>30525-89-4&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Propidium iodide</td>
			<td>Sigma-Aldrich</td>
			<td>P4864</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Iodine</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>7681-82-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Thyroglobulin</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>T1126</td>
			<td></td>
		</tr>
		<tr>
			<td>Thyroglobulin&#160; ELISA Kit</td>
			<td>Thermo Scientific</td>
			<td>EHTGX5</td>
			<td></td>
		</tr>
		<tr>
			<td>TSH ELISA</td>
			<td>Calbiotech</td>
			<td>DKO200</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>macklin</td>
			<td>1330-20-7</td>
			<td></td>
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generation of a mouse spontaneous autoimmune thyroiditis model

In recent years, Hashimoto's thyroiditis (HT) has become the most common autoimmune thyroid disease. It is characterized by lymphocyte infiltration and the detection of specific serum autoantibodies. Although the potential mechanism is still not clear, the risk of Hashimoto's thyroiditis is related to genetic and environmental factors. At present, there are several types of models of autoimmune thyroiditis, including experimental autoimmune thyroiditis (EAT) and spontaneous autoimmune thyroiditis (SAT).
EAT in mice is a common model for HT, which is immunized with lipopolysaccharide (LPS) combined with thyroglobulin (Tg) or supplemented with complete Freund's adjuvant (CFA). The EAT mouse model is widely established in many types of mice. However, the disease progression is more likely associated with the Tg antibody response, which may vary in different experiments.
SAT is also widely used in the study of HT in the NOD.H-2h4 mouse. The NOD.H2h4 mouse is a new strain obtained from the cross of the nonobese diabetic (NOD) mouse with the B10.A(4R), which is significantly induced for HT with or without feeding iodine. During the induction, the NOD.H-2h4 mouse has a high level of TgAb accompanied by lymphocyte infiltration in the thyroid follicular tissue. However, for this type of mouse model, there are few studies to comprehensively evaluate the pathological process during the induction of iodine.
A SAT mouse model for HT research is established in this study, and the pathologic changing process is evaluated after a long period of iodine induction. Through this model, researchers can better understand the pathological development of HT and screen new treatment methods for HT.

The histological changes were strikingly different in female and males, the duration of iodine intake, and the solution of NaI. As shown in Figure 1, ~10% of NOD.H-2h4 mice developed SAT even without iodine induction at the age of 24 weeks, and all the mice eventually developed thyroiditis. When given regular water, there was no significant difference in the histological changes between males and females. The addition of NaI to the drinking water accelerated the development of thyroiditis. I...

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Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model

Several types of animal models of Hashimoto's thyroiditis have been established, as has spontaneous autoimmune thyroiditis in the NOD mouse. H-2h4 mice are a simple and reliable model for HT induction. This article describes this approach and evaluates the pathological process for a better understanding of the SAT murine model.

In recent years, Hashimoto's thyroiditis (HT) has become the most common autoimmune thyroid disease. It is characterized by lymphocyte infiltration and ...

Autoimmune thyroiditis, the outcome of disease that seriously affects human health. Although there are currently no effective treatment, the animal models provide a platform for studying this disease. This model is easy to construct with simple steps and a high success rate.It stimulates the autoimmune process with verifying the relationship between our dying and the disease severity. To begin, fix the mouse on the dissection table with pins. To prepare the thyroid tissue sample, dissect the chest wall and expose the heart.Cut open the right atrium and inject saline intravenously into the left ventricle, using a 20 milliliter syringe until the tissue turns white. Sterilize the neck with 75%ethanol. Using tissue scissors, cut the mouse skin along the neck, midline from the top of the sternum to the mandible, to fully expose the neck tissue.Locate the submandibular glands and anterior trachea muscle and separate them using ophthalmic scissors and forceps. Once the trachea and thyroid cartilage are exposed, remove tissue under the cartilage using curved forceps. After picking up the cartilage and trachea, cut the distal lens of the cartilage and proximal lens of the trachea.Then remove the thyroid gland and the trachea. The HE staining showed pathological changes in the thyroid gland, under iodine intake. All the mice developed thyroiditis, even without iodine induction.However, accelerated development of thyroiditis was observed with increasing sodium iodide concentration. Female mice tend to develop more severe thyroiditis than males under the same condition, but differences in the severity of lymphocytic infiltration were negligible. Thyroglobulin antibody levels were not significantly increased in plain water at 16 weeks of age.Whereas, sodium iodide supplementation exhibited a comparable rise in antibody levels in male and female mice. The thyroid peroxidase antibodies were found at 24 weeks in regular water;and female mice showed higher levels than males at 72 weeks. However, the 0.05%sodium iodide edition showed the highest antibodies at 64 weeks.In addition, TSH levels were higher in male than female NODH2 H4 mice;with or without iodide supplementation. Furthermore, the T4 levels showed decreasing tendency to induction time in iodine water. It should be noted that 0.05%sodium at the most suitable concentration to reduce the salt model.Also, more care should be taken while isolating specimens. Because the mouse cell is relatively small and easily damaged. The establishment of the animal model is helpful for the study of the path genocide and the treatment of autoimmune cell status.And provides a basis for the development of new treatment method.

Research

JoVE Journal

Immunology and Infection

Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis

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Induction of Experimental Autoimmune Hypophysitis in SJL Mice

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Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model

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Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation

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Simple and Efficient Production and Purification of Mouse Myelin Oligodendrocyte Glycoprotein for Experimental Autoimmune Encephalomyelitis Studies

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Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291

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Clostridium difficile is an anaerobic, gram-positive, spore-forming enteric pathogen that is associated with increasing morbidity and mortality and ...

A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue

Dry eye disease is a very common condition that causes morbidity and healthcare burden and decreases the quality of life. There is a need for a suitable ...

Influenza A Virus Studies in a Mouse Model of Infection

Influenza viruses cause over 500,000 deaths worldwide1 and are associated with an annual cost of 12 - 14 billion USD in the United States alone ...

High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model

Type 1 Diabetes (T1D) is characterized by islet-specific autoimmunity leading to beta cell destruction and absolute loss of insulin production. In the ...