Name: generation of a mouse spontaneous autoimmune thyroiditis model
Uploaded: 2023-03-17T14:05:00
Description: Watch this Scientific Journal Video about Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model at JoVE.com

인간 TPO에 대한 마우스 단일클론 항체(양성 대조군으로 사용됨)는 Dr. P. Carayon 및 Dr. J. Ruf(프랑스 마르세유)에 의해 제공되었습니다. 저자는이 연구의 모든 참가자와 우리 연구팀 구성원에게 감사드립니다. 이 작업은 중국 쓰촨 대학교 서중국 병원의 박사후 유지 기금(2020HXBH057) 및 쓰촨성 과학 기술 지원 프로그램(프로젝트 번호 2021YFS0166)의 보조금으로 부분적으로 지원되었습니다

아래에 설명된 프로토콜은 쓰촨 대학교의 기관 동물 관리 및 사용 위원회에서 제정한 관리 및 사용 지침에 의해 승인되었습니다.
1. 준비
<ol><li>모든 마우스를 12시간 명암 주기(각각 오전 07:00 및 오후 07:00에 시작)에서 특정 병원체가 없는 조건에 수용합니다. 실내 온도를 22°C로 유지합니다. 매주 침구 재료를 교체하십시오. 적절한 양의 표준 설치류 차우와 물을...

<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. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+thyroiditis+in+rabbits,+guinea+pigs+and+dogs,+following+immunization+with+thyroid+extracts+of+their+own+and+of+heterologous+species.">Experimental thyroiditis in rabbits, guinea pigs and dogs, following immunization with thyroid extracts of their own and of heterologous species.</a> The American Journal of Pathology. 36 (2), 213-239 (1960).</li><li>Alexopoulos, H., Dalakas, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+immunobiology+of+autoimmune+encephalitides.">The immunobiology of autoimmune encephalitides.</a> Journal of Autoimmunity. 104, 102339 (2019).</li><li>Noviello, C. M., Kreye, J., Teng, J., Pr&#252;ss, H., Hibbs, R. 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. 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=Superiority+of+thyroid+peroxidase+DNA+over+protein+immunization+in+replicating+human+thyroid+autoimmunity+in+HLA-DRB1*0301+(DR3)+transgenic+mice.">Superiority of thyroid peroxidase DNA over protein immunization in replicating human thyroid autoimmunity in HLA-DRB1*0301 (DR3) transgenic mice.</a> Clinical and Experimental Immunology. 137 (3), 503-512 (2004).</li><li>Akeno, N., 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=IFN-&#945;+mediates+the+development+of+autoimmunity+both+by+direct+tissue+toxicity+and+through+immune+cell+recruitment+mechanisms.">IFN-&#945; mediates the development of autoimmunity both by direct tissue toxicity and through immune cell recruitment mechanisms.</a> Journal of Immunology. 186 (8), 4693-4706 (2011).</li></ol>

HT는 갑상선에 침투한 림프구로 인한 자가면역계 장애로 인해 발생하며, 갑상선 기능을 더욱 손상시키면서 갑상선 특이적 항체를 생성합니다. HT 환자의 혈청 TSH, TgAb 및 TPOAb 수치는 유의하게 상승했다27. 현재 자가면역 갑상선염의 병인을 연구하기 위해 두 가지 주요 종류의 쥐 모델이 널리 사용됩니다: EAT와 SAT29. EAT 마우스는 생체 내에서 비정상적인 면?...

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

하시모토 갑상선염(Hashimoto's thyroiditis, HT)은 만성 림프구성 갑상선염 또는 자가면역성 갑상선염으로도 알려져 있으며, 1912년에 처음 보고되었다1. HT는 림프구 침윤과 갑상선 여포 조직 손상을 특징으로 합니다. 실험실 검사는 주로 항갑상선글로불린 항체(TgAb) 및 항갑상선 과산화효소 항체(TPOAb)를 포함한 갑상선 특이적 항체의 증가로 나타납니다.2. HT의 발병률은 0.4%-1.5% 범위로 모든 갑상선 질환의 20%-25%를 차지하며 이 값은 최근 몇 년 동안 증가했습니다3. 또한 많은 연구에서 HT가 유두상 갑상선 암종(PTC)의 종양 발생 및 재발과 관련이 있다고 보고했습니다.4,5; 잠재적 메커니즘은 여전히 논란의 여지가 있습니다. 자가면역성 갑상선염은 여성 불임의 중요한 요인이기도하다 6. 따라서 HT의 발병기전이 명확해야 하며, 이를 위해서는 안정적이고 단순한 동물 모델이 필수적입니다.
HT의 병인을 연구하기 위해 본 연구에서 실험적 자가면역 갑상선염(EAT)과 자발성 자가면역 갑상선염(SAT)을 포함하여 두 가지 주요 종류의 쥐 모델이 사용되었습니다 7,8. 감수성 마우스를 특정 갑상선 항원(조갑상선, 정제된 티로글로불린[TG], 갑상선 과산화효소[TPO], 재조합 TPO 엑토도메인 및 선택된 TPO 펩티드 포함)으로 면역화하여 EAT 쥐 모델을 확립했습니다. 또한, 지질다당류(LPS), 완전 프로인트 보조제(CFA) 및 다른 특이한 보조제를 포함하는 보조제는 또한 면역 관용(9,10,11,12,13,14,15,16,17)을 분해하기 위해 면역화 동안 사용된다.
SAT 모델은 NOD를 기반으로 하는 자가면역성 갑상선염의 자발적 발달을 연구하는 중요한 모델입니다. H-2h4 마우스. 고개를 끄덕입니다. H-2h4 마우스는 NOD와 B10의 교배로부터 얻은 새로운 균주이다. A(4R) 마우스, 자가면역 갑상선염 감수성 유전자 IAk18,19를 사용하여 NOD에 대한 다중 백크로스. 끄덕이다. H-2h4 마우스는 당뇨병이 발생하지 않지만 자가면역 갑상선염과 쇼그렌 증후군(SS) 발병률이 높습니다19. 연구에 따르면 세포 내 접착 분자-1(ICAM-1)은 NOD의 갑상선 조직에서 고도로 발현됩니다. 3-4주령의 H-2H4 마우스. 또한, 요오드 섭취가 증가함에 따라 티로 글로불린 분자의 면역 원성이 향상되어 단핵구 침윤 과정에서 중요한 역할을하는 ICAM-1의 발현을 더욱 상향 조절한다21. 이 모델은 요오드 용량과 질병 중증도 사이의 관계를 확인하면서 자가면역 과정을 시뮬레이션합니다. 확립 된 방법은 안정적이며 성공 확률이 높습니다. SAT 모델은 수년 동안 자가면역성 갑상선염을 유발하는 데 적용되어 왔으며 자가면역성 갑상선염의 발병기전을 연구하는 효과적인 방법이기도 합니다. 그러나 현재 EAT 모델의 구성 방법은 더 복잡하고 비용이 많이 듭니다. 다른 실험실은 다른 예방 접종 방법과 주사 부위를 사용합니다. 또한, 유전적 배경이 다른 마우스는 유도 속도가 다르기 때문에 강력한 메커니즘을 밝히기 위해서는 추가 연구가 필요합니다.
그러나 SAT 모델에서 갑상선염의 발병은 요오드화 나트륨, 성적 이형성 및 양육 조건과 관련이 있습니다. SAT 모델에서자가 면역 갑상선염의 적절한 절차를 밝히기 위해이 기사에서는 다양한 조건에서자가 면역 갑상선염을 유도하는 방법을 설명했습니다. 또한, 이 질환의 여러 단계에서 자가면역 갑상선염의 발병기전 및 면역학적 진행을 연구할 수 있습니다.

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			<td>Freund&#8217;s Adjuvant, Incomplete&#160;</td>
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			<td>Goat anti-Mouse IgG&#160;</td>
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			<td>Midazolam solution&#160;</td>
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			<td>Mouse/rat thyroxine (T4) ELISA</td>
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			<td>Propidium iodide</td>
			<td>Sigma-Aldrich</td>
			<td>P4864</td>
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			<td>Sodium Iodine</td>
			<td>Sigma-Aldrich&#160;</td>
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			<td>Thyroglobulin</td>
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			<td>Thyroglobulin&#160; ELISA Kit</td>
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			<td>EHTGX5</td>
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			<td>Calbiotech</td>
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			<td>Xylene</td>
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			<td>1330-20-7</td>
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generation of a mouse spontaneous autoimmune thyroiditis model

최근 몇 년 동안 하시모토 갑상선염(HT)은 가장 흔한 자가면역 갑상선 질환이 되었습니다. 그것은 림프구 침윤과 특정 혈청자가 항체의 검출을 특징으로합니다. 잠재적인 메커니즘은 아직 명확하지 않지만 하시모토 갑상선염의 위험은 유전적 및 환경적 요인과 관련이 있습니다. 현재 실험적 자가면역 갑상선염(EAT) 및 자발성 자가면역 갑상선염(SAT)을 포함하여 여러 유형의 자가면역 갑상선염 모델이 있습니다.
생쥐의 EAT는 티로글로불린(Tg)과 결합된 지질다당류(LPS)로 면역화되거나 완전 프로인트 보조제(CFA)로 보충되는 HT의 일반적인 모델입니다. EAT 마우스 모델은 많은 유형의 마우스에서 널리 확립되어 있습니다. 그러나, 질환 진행은 Tg 항체 반응과 연관될 가능성이 더 높으며, 이는 상이한 실험에서 다를 수 있다.
SAT는 NOD의 HT 연구에도 널리 사용됩니다. H-2h4 마우스. 고개를 끄덕입니다. H2h4 마우스는 B10과 함께 비만하지 않은 당뇨병(NOD) 마우스의 교배로부터 얻은 새로운 균주이다. A(4R), 이는 요오드 공급 유무에 관계없이 HT에 대해 유의하게 유도됩니다. 유도하는 동안 NOD. H-2h4 마우스는 갑상선 여포 조직에 림프구 침윤을 동반하는 높은 수준의 TgAb를 가지고 있습니다. 그러나 이러한 유형의 마우스 모델의 경우 요오드 유도 동안 병리학적 과정을 종합적으로 평가하는 연구는 거의 없습니다.
본 연구에서는 HT 연구를 위한 SAT 마우스 모델을 확립하고, 장기간의 요오드 유도 후 병리학적 변화 과정을 평가한다. 이 모델을 통해 연구자들은 HT의 병리학적 발달을 더 잘 이해하고 HT에 대한 새로운 치료 방법을 선별할 수 있습니다.

조직학적 변화는 여성과 남성, 요오드 섭취 기간, NaI 용액에서 현저하게 달랐습니다. 그림 1에서 볼 수 있듯이 NOD의 ~10%. H-2h4 마우스는 24주령에 요오드 유도 없이도 SAT가 발생했고, 결국 모든 마우스에서 갑상선염이 발생했다. 일반 물을 투여했을 때 남성과 여성의 조직학적 변화에는 큰 차이가 없었습니다. 식수에 NaI를 추가하면 갑상선염의 발병이 가속화되었습니다. 0.005%, ...

Watch this Scientific Journal Video about Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model at JoVE.com

Generation of a Mouse Spontaneous Autoimmune Thyroiditis Model

하시모토 갑상선염의 여러 유형의 동물 모델이 확립되었으며, NOD 마우스의 자발성 자가면역 갑상선염도 확립되었습니다. H-2h4 마우스는 HT 유도를 위한 간단하고 신뢰할 수 있는 모델입니다. 이 기사에서는 이 접근 방식을 설명하고 SAT 쥐 모델을 더 잘 이해하기 위해 병리학적 과정을 평가합니다.

마우스 자발성 자가면역 갑상선염 모델 생성

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

자가 면역 갑상선염은 인체 건강에 심각한 영향을 미치는 질병의 결과입니다. 현재 효과적인 치료법은 없지만 동물 모델은 이 질병을 연구하기 위한 플랫폼을 제공합니다. 이 모델은 간단한 단계와 높은 성공률로 쉽게 구성할 수 있습니다.그것은 우리의 죽음과 질병의 중증도 사이의 관계를 확인하여자가 면역 과정을 자극합니다. 시작하려면 핀으로 해부 테이블에 마우스를 고정하십시오. 갑상선 조직 샘플을 준비하려면 흉벽을 해부하고 심장을 노출시킵니다.우심방을 잘라내고 조직이 하얗게 변할 때까지 20밀리리터 주사기를 사용하여 좌심실에 식염수를 정맥 주사합니다. 75 % 에탄올로 목을 소독하십시오. 조직 가위를 사용하여 목을 따라 마우스 피부를 흉골 상단에서 하악까지 정중선으로 절단하여 목 조직을 완전히 노출시킵니다.턱밑샘과 전기관근을 찾아 안과용 가위와 집게를 사용하여 분리합니다. 기관과 갑상선 연골이 노출되면 구부러진 집게를 사용하여 연골 아래 조직을 제거합니다. 연골과 기관을 채취한 후 연골의 원위 수정체와 기관의 근위 수정체를 절단합니다.그런 다음 갑상선과 기관을 제거하십시오. HE 염색은 요오드 섭취 하에서 갑상선의 병리학적 변화를 보여주었습니다. 모든 생쥐는 요오드 유도 없이도 갑상선염을 일으켰습니다.그러나 요오드화 나트륨 농도가 증가함에 따라 갑상선염의 가속화 된 발달이 관찰되었습니다. 암컷 마우스는 같은 조건에서 수컷보다 더 심한 갑상선염이 발생하는 경향이 있지만 림프구 침윤의 중증도의 차이는 무시할 수 있었습니다. 티로 글로불린 항체 수치는 16 주령의 일반 물에서 유의하게 증가하지 않았습니다.반면, 요오드화 나트륨 보충제는 수컷과 암컷 마우스에서 항체 수치가 비슷한 상승을 보였습니다. 갑상선 과산화효소 항체는 일반 물에서 24주에 발견되었으며, 암컷 쥐는 72주에 수컷보다 높은 수치를 보였다. 그러나 0.05% 요오드화나트륨 판은 64주에 가장 높은 항체를 보였다.또한, TSH 수치는 요오드화물 보충 유무에 관계없이 암컷 NODH2 H4 마우스보다 수컷에서 더 높았다. 또한, T4 수준은 요오드 물에서 유도 시간이 감소하는 경향을 보였다. 0.05%나트륨은 소금 모델을 감소시키는 데 가장 적합한 농도입니다.또한 표본을 분리하는 동안 더 많은 주의를 기울여야 합니다. 마우스 셀이 상대적으로 작고 쉽게 손상되기 때문입니다. 상기 동물모델의 확립은 자가면역세포 상태의 대량 학살 경로 및 치료 경로에 대한 연구에 도움이 된다.그리고 새로운 치료 방법의 개발을 위한 기초를 제공합니다.

generation-of-a-mouse-spontaneous-autoimmune-thyroiditis-model

Research

JoVE Journal

Immunology and Infection

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

Autoimmune hypophysitis is a chronic inflammation of the pituitary gland caused or accompanied by autoimmunity1. It has traditionally been considered a rare disease but reporting has increased markedly in recent years. Hypophysitis, in fact, develops not uncommonly as a "side effect" in cancer patients treated with antibodies that block inhibitory receptors expressed on T lymphocytes, such as CTLA-42 and PD-1 receptors. Autoimmune hypophysitis can be induced experimentally by injecting mice with pituitary proteins mixed with an adjuvant3. In this video article we demonstrate how to extract proteins from mouse pituitary glands and how to prepare them in a form suitable for inducing autoimmune hypophysitis in SJL mice.

Autoimmune hypophysitis can be reproduced in mice by injecting an extract of mouse pituitary proteins.

실험 면역 Hypophysitis의 유도를위한 마우스 뇌하수체 Immunogen의 준비

Autoimmune hypophysitis is a chronic inflammation of the pituitary gland caused or accompanied by autoimmunity1.  It has traditionally been considered a ...

연구

면역학 및 감염병학

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse models allow us to study how diseases unfold, often providing a good replica of the same processes occurring in humans. For some autoimmune diseases, like type 1A diabetes, there are models (the NOD mouse) that spontaneously develop a disease similar to the human counterpart. For many other autoimmune diseases, however, the model needs to be induced experimentally. A common approach in this regard is to inject the mouse with a dominant antigen derived from the organ being studied. For example, investigators interested in autoimmune thyroiditis inject mice with thyroglobulin2, and those interested in myasthenia gravis inject them with the acetylcholine receptor3. If the autoantigen for a particular autoimmune disease is not known, investigators inject a crude protein extract from the organ targeted by the autoimmune reaction. For autoimmune hypophysitis, the pathogenic autoantigen(s) remain to be identified4, and thus a crude pituitary protein preparation is used. In this video article we demonstrate how to induce experimental autoimmune hypophysitis in SJL mice.

This video shows how to induce autoimmune hypophysitis in SJL mice and how to assess its severity by histopathology.

SJL 마우스의 실험적 면역 Hypophysitis의 유도

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse ...

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

The Trypanosoma cruzi acute infections acquired in infancy and childhood seem asymptomatic, but approximately one third of the chronically infected cases show Chagas disease up to three decades or later. Autoimmunity and parasite persistence are competing theories to explain the pathogenesis of Chagas disease 1, 2. To separate roles played by parasite persistence and autoimmunity in Chagas disease we inoculate the T. cruzi in the air chamber of fertilized eggs. The mature chicken immune system is a tight biological barrier against T. cruzi and the infection is eradicated upon development of its immune system by the end of the first week of growth 3. The chicks are parasite-free at hatching, but they retain integrated parasite mitochondrial kinetoplast DNA (kDNA) minicircle within their genome that are transferred to their progeny. Documentation of the kDNA minicircle integration in the chicken genome was obtained by a targeted prime TAIL-PCR, Southern hybridizations, cloning, and sequencing 3, 4. The kDNA minicircle integrations rupture open reading frames for transcription and immune system factors, phosphatase (GTPase), adenylate cyclase and phosphorylases (PKC, NF-Kappa B activator, PI-3K) associated with cell physiology, growth, and differentiation 3, 5-7, and other gene functions. Severe myocarditis due to rejection of target heart fibers by effectors cytotoxic lymphocytes is seen in the kDNA mutated chickens, showing an inflammatory cardiomyopathy similar to that seen in human Chagas disease. Notably, heart failure and skeletal muscle weakness are present in adult chickens with kDNA rupture of the dystrophin gene in chromosome 1 8. Similar genotipic alterations are associated with tissue destruction carried out by effectors CD45+, CD8&gamma;&delta;+, CD8&alpha; lymphocytes. Thus this protozoan infection can induce genetically driven autoimmune disease.

The inoculation of Trypanosoma cruzi in fertile eggs prior to incubation renders the parasite kDNA minicircle integration in embryo cells genome. Crossbreeding reveals the vertical transfer of the mutations to progeny. The kDNA integrates into coding regions at several chromosomes and the chickens die with an inflammatory autoimmune heart disease.

기생충은 치킨 모델에서 유전 힘입어 면역 Chagas의 심장 질환을 유발

The Trypanosoma cruzi acute infections acquired in infancy and childhood seem asymptomatic, but approximately one third of the chronically infected cases ...

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

Dissociating murine skin into a single cell suspension is essential for downstream cellular analysis such as the characterization of infiltrating immune cells in rodent models of skin inflammation. Here, we describe a protocol for the digestion of mouse skin in a nutrient-rich solution with collagenase D, followed by separation of hematopoietic cells using a discontinuous density gradient. Cells thus obtained can be used for in vitro studies, in vivo transfer, and other downstream cellular and molecular analyses including flow cytometry. This protocol is an effective and economical alternative to expensive mechanical dissociators, specialized separation columns, and harsher trypsin- and dispase-based digestion methods, which may compromise cellular viability or density of surface proteins relevant for phenotypic characterization or cellular function. As shown here in our representative data, this protocol produced highly viable cells, contained specific immune cell subsets, and had no effect on integrity of common surface marker proteins used in flow cytometric analysis.

This protocol describes enzymatic digestion of mouse skin in nutrient-rich medium followed by gradient separation to isolate leukocytes. Cells thus derived can be used for diverse downstream applications. This is an effective, economical, and improved alternative to tissue dissociation machines and harsher trypsin and dispase-based tissue digestion protocols.

효소 다이제스트 및 그라데이션 분리를 사용하여 마우스 피부에서 침투 백혈구의 분리

Dissociating murine skin into a single cell suspension is essential for downstream cellular analysis such as the characterization of infiltrating immune ...

Generation and Identification of GM-CSF Derived Alveolar-like Macrophages and Dendritic Cells From Mouse Bone Marrow

Macrophages and dendritic cells (DCs) are innate immune cells found in tissues and lymphoid organs that play a key role in the defense against pathogens. However, they are difficult to isolate in sufficient numbers to study them in detail, therefore, in vitro models have been developed. In vitro cultures of bone marrow-derived macrophages and dendritic cells are well-established and valuable methods for immunological studies. Here, a method for culturing and identifying both DCs and macrophages from a single culture of primary mouse bone marrow cells using the cytokine granulocyte macrophage colony-stimulating factor (GM-CSF) is described. This protocol is based on the established procedure first developed by Lutz et al. in 1999 for bone marrow-derived DCs. The culture is heterogeneous, and MHCII and fluoresceinated hyaluronan (FL-HA) are used to distinguish macrophages from immature and mature DCs. These GM-CSF derived macrophages provide a convenient source of in vitro derived macrophages that closely resemble alveolar macrophages in both phenotype and function.

Bone marrow cells cultured with granulocyte macrophage colony stimulating factor (GM-CSF) generate a heterogeneous culture containing macrophages and dendritic cells (DCs). This method highlights using MHCII and hyaluronan (HA) binding to differentiate macrophages from the DCs in the GM-CSF culture. Macrophages in this culture have many similarities to alveolar macrophages.

마우스 골수에서 생성 및 GM-CSF의 식별 파생 폐포와 같은 대 식세포와 수지상 세포

Macrophages and dendritic cells (DCs) are innate immune cells found in tissues and lymphoid organs that play a key role in the defense against pathogens. ...

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

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS), thought to occur as a result of autoimmune responses targeting myelin. Experimental autoimmune encephalomyelitis (EAE) is the most common animal model of CNS autoimmune disease, and is typically induced via immunization with short peptides representing immunodominant CD4+ T cell epitopes of myelin proteins. However, B cells recognize unprocessed protein directly, and immunization with short peptide does not activate B cells that recognize the native protein. As recent clinical trials of B cell-depleting therapies in MS have suggested a role for B cells in driving disease in humans, there is an urgent need for animal models that incorporate B cell-recognition of autoantigen. To this end, we have generated a new fusion protein containing the extracellular domain of the mouse version of myelin oligodendrocyte glycoprotein (MOG) as well as N-terminal fusions of a His-tag for purification purposes and the thioredoxin protein to improve solubility (MOGtag). A tobacco etch virus (TEV) protease cleavage site was incorporated to allow the removal of all tag sequences, leaving only the pure MOG1-125 extracellular domain. Here, we describe a simple protocol using only standard laboratory equipment to produce large quantities of pure MOGtag or MOG1-125. This protocol consistently generates over 200 mg of MOGtag protein. Immunization with either MOGtag or MOG1-125 generates an autoimmune response that includes pathogenic B cells that recognize the native mouse MOG.

We describe a simple protocol using only basic lab equipment to generate and purify large quantities of a fusion protein that contains mouse Myelin Oligodendrocyte Glycoprotein. This protein can be used to induce experimental autoimmune encephalomyelitis driven by both T and B cells.

실험자가 면역 뇌척수염 연구 간단하고 효율적인 생산 및 마우스 미엘린 희소 돌기 아교 세포 당 단백질의 정제

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS), thought to occur as a result of autoimmune responses ...

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

Clostridium difficile is an anaerobic, gram-positive, spore-forming enteric pathogen that is associated with increasing morbidity and mortality and consequently poses an urgent threat to public health. Recurrence of a C. difficile infection (CDI) after successful treatment with antibiotics is high, occurring in 20-30% of patients, thus necessitating the discovery of novel therapeutics against this pathogen. Current animal models of CDI result in high mortality rates and thus do not approximate the chronic, insidious disease manifestations seen in humans with CDI. To evaluate therapeutics against C. difficile, a mouse model approximating human disease utilizing a clinically-relevant strain is needed. This protocol outlines the cefoperazone mouse model of CDI using a clinically-relevant and genetically-tractable strain, R20291. Techniques for clinical disease monitoring, C. difficile bacterial enumeration, toxin cytotoxicity, and histopathological changes throughout CDI in a mouse model are detailed in the protocol. Compared to other mouse models of CDI, this model is not uniformly lethal at the dose administered, allowing for the observation of a prolonged clinical course of infection concordant with the human disease. Therefore, this cefoperazone mouse model of CDI proves a valuable experimental platform to assess the effects of novel therapeutics on the amelioration of clinical disease and on the restoration of colonization resistance against C. difficile.

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

This protocol outlines the cefoperazone mouse model of Clostridium difficile infection (CDI) using a clinically-relevant and genetically-tractable strain, R20291. Emphasis on clinical disease monitoring, C. difficile bacterial enumeration, toxin cytotoxicity, and histopathological changes throughout CDI in a mouse model are detailed in the protocol.

의 Cefoperazone 처리 된 마우스 모델 임상 관련<em&gt; 클로스 트리 디움 디피</em&gt; 스트레인 R20291

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 dry eye animal model to test novel therapeutics to treat autoimmune dry eye conditions. This protocol describes a chronic autoimmune dry eye rat model. Lewis rats were immunized with an emulsion containing lacrimal gland extract, ovalbumin, and complete Freund&#39;s adjuvant. A second immunization with the same antigens in incomplete Freund&#39;s adjuvant was administered two weeks later. These immunizations were administered subcutaneously at the base of the tail. To boost the immune response at the ocular surface and lacrimal glands, lacrimal gland extract and ovalbumin were injected into the forniceal subconjunctiva and lacrimal glands 6 weeks after the first immunization. The rats developed dry eye features, including reduced tear production, decreased tear stability, and increased corneal damage. Immune profiling by flow cytometry showed a preponderance of CD3+ effector memory T cells in the eyeball.

This report describes a method to induce chronic experimental autoimmune dry eye in Lewis rats through immunization with an emulsion of rat lacrimal gland extract, ovalbumin, and complete Freund's adjuvant, followed by the injection of lacrimal gland extract and ovalbumin into the forniceal subconjunctiva and lacrimal glands six weeks later.

안구 조직에서 이펙터 기억 T 세포의 증가를 동반 한 만성자가 면역 건식 ​​쥐 모델

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 considering direct medical and hospitalization expenses and work absenteeism2. Animal models are crucial in Influenza A virus (IAV) studies to evaluate viral pathogenesis, host-pathogen interactions, immune responses, and the efficacy of current and/or novel vaccine approaches as well as antivirals. Mice are an advantageous small animal model because their immune system is evolutionarily similar to that found in humans, they are available from commercial vendors as genetically identical subjects, there are multiple strains that can be exploited to evaluate the genetic basis of infections, and they are relatively inexpensive and easy to manipulate. To recapitulate IAV infection in humans via the airways, mice are first anesthetized prior to intranasal inoculation with infectious IAVs under proper biosafety containment. After infection, the pathogenesis of IAVs is determined by monitoring daily the morbidity (body weight loss) and mortality (survival) rate. In addition, viral pathogenesis can also be evaluated by assessing virus replication in the upper (nasal mucosa) or lower (lungs) respiratory tract of infected mice. Humoral responses upon IAV infection can be rapidly evaluated by non-invasive bleeding and secondary antibody detection assays aimed at detecting the presence of total or neutralizing antibodies. Here, we describe the common methods used to infect mice intranasally (i.n) with IAV and evaluate pathogenesis, humoral immune responses and protection efficacy.

Influenza A viruses (IAVs) are important human respiratory pathogens. To understand the pathogenicity of IAVs and to perform preclinical testing of novel vaccine approaches, animal models mimicking human physiology are required. Here, we describe techniques to evaluate IAV pathogenesis, humoral responses and vaccine efficacy using a mouse model of infection.

감염 마우스 모델에서 인플루엔자 A 바이러스 연구

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 ...

A Controlled Mouse Model for Neonatal Polymicrobial Sepsis

Neonatal sepsis remains a global burden. A preclinical model to screen effective prophylactic or therapeutic interventions is needed. Neonatal mouse polymicrobial sepsis can be induced by injecting cecal slurry intraperitoneally into day of life 7 mice and monitoring them for the following week. Presented here are the detailed steps necessary for the implementation of this neonatal sepsis model. This includes making a homogeneous cecal slurry stock, diluting it to a weight- and litter-adjusted dose, an outline of the monitoring schedule, and a definition of observed health categories used to define humane endpoints. The generation of a homogeneous cecal slurry stock from pooled donors allows for the administration into many litters over time, reducing the variation between donors, and preventing the use of potentially toxic glycerol. The monitoring strategy used allows for the anticipation of survival outcome and the identification of mice that would later progress to death, allowing for an earlier identification of the humane endpoint. Two main behavioral features are used to define the health scores, namely, the ability of the neonatal mice to right themselves when placed on their back and their level of mobility. These criteria could potentially be applied to address humane endpoints in other studies of neonatal disease in mice, as long as a pilot study is performed to confirm accuracy. In conclusion, this approach provides a standardized method to model newborn sepsis in mice, while providing resources to assess animal welfare used to define early humane endpoints for challenged animals.

This protocol provides the necessary steps to establish and evaluate neonatal sepsis in 7-day-old mice.

신생아 Polymicrobial 패 혈 증에 대 한 제어 마우스 모델

Neonatal sepsis remains a global burden. A preclinical model to screen effective prophylactic or therapeutic interventions is needed. Neonatal mouse ...