{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:12:50Z","timestamp":1760188370402},"reference-count":52,"publisher":"Oxford University Press (OUP)","issue":"3","license":[{"start":{"date-parts":[[2004,5,4]],"date-time":"2004-05-04T00:00:00Z","timestamp":1083628800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2004,5,4]]},"abstract":"<jats:title>SUMMARY<\/jats:title><jats:p>Intramuscular injection with plasmid DNA encoding the human thyrotropin receptor (TSHR) has been known to elicit symptoms of Graves\u2019 disease (GD) in outbred but not inbred mice. In this study, we have examined, firstly, whether intradermal (i.d.) injection of TSHR DNA can induce hyperthyroidism in BALB\/c mice and, secondly, whether coinjection of TSHR- and cytokine-producing plasmids can influence the outcome of disease. Animals were i.d. challenged at 0, 3 and 6\u2003weeks with TSHR DNA and the immune response was assessed at the end of the 8th or 10th week. In two experiments, a total of 10 (67%) of 15 mice developed TSHR-specific antibodies as assessed by flow cytometry. Of these, 4 (27%) mice had elevated thyroxine (TT4) levels and goitrous thyroids with activated follicular epithelial cells but no evidence of lymphocytic infiltration. At 10\u2003weeks, thyroid-stimulating antibodies (TSAb) were detected in two out of the four hyperthyroid animals. Interestingly, in mice that received a coinjection of TSHR- and IL-2- or IL-4-producing plasmids, there was no production of TSAbs and no evidence of hyperthyroidism. On the other hand, coinjection of DNA plasmids encoding TSHR and IL-12 did not significantly enhance GD development since two out of seven animals became thyrotoxic, but had no goitre. These results demonstrate that i.d. delivery of human TSHR DNA can break tolerance and elicit GD in inbred mice. The data do not support the notion that TSAb production is Th2-dependent in murine GD but they also suggest that codelivery of TSHR and Th1-promoting IL-12 genes may not be sufficient to enhance disease incidence and\/or severity in this model.<\/jats:p>","DOI":"10.1111\/j.1365-2249.2004.02483.x","type":"journal-article","created":{"date-parts":[[2004,5,14]],"date-time":"2004-05-14T14:46:12Z","timestamp":1084545972000},"page":"413-422","source":"Crossref","is-referenced-by-count":14,"title":["Induction of hyperthyroidism in mice by intradermal immunization with DNA encoding the thyrotropin receptor"],"prefix":"10.1093","volume":"136","author":[{"given":"K","family":"BARRETT","sequence":"first","affiliation":[{"name":"Division of Endocrinology, Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E","family":"LIAKATA","sequence":"additional","affiliation":[{"name":"Immunology Laboratory, Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P V","family":"RAO","sequence":"additional","affiliation":[{"name":"Division of Medicine, GKT School of Medicine, London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P F","family":"WATSON","sequence":"additional","affiliation":[{"name":"Clinical Sciences Centre, University of Sheffield, Sheffield, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A P","family":"WEETMAN","sequence":"additional","affiliation":[{"name":"Clinical Sciences Centre, University of Sheffield, Sheffield, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P","family":"LYMBERI","sequence":"additional","affiliation":[{"name":"Immunology Laboratory, Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J P","family":"BANGA","sequence":"additional","affiliation":[{"name":"Division of Medicine, GKT School of Medicine, London, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G","family":"CARAYANNIOTIS","sequence":"additional","affiliation":[{"name":"Division of Endocrinology, Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2004,5,5]]},"reference":[{"key":"2022011223181945100_b1","first-page":"219","volume-title":"Werner & Ingbar's The Thyroid: a fundamental and clinical text","author":"Rapoport","year":"2000"},{"key":"2022011223181945100_b2","first-page":"518","volume-title":"Werner & Ingbar's The Thyroid: a fundamental and clinical text","author":"Davies","year":"2000"},{"key":"2022011223181945100_b3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1530\/eje.0.1420001","article-title":"Animal models of Graves\u2019 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model","volume":"135","author":"Costagliola","year":"1994","journal-title":"Endocrinology"},{"key":"2022011223181945100_b7","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1006\/bbrc.1994.1332","article-title":"Induction of thyrotropin receptor (TSH-R) autoantibodies and thyroiditis in mice immunised with the recombinant TSH-R","volume":"199","author":"Costagliola","year":"1994","journal-title":"Biochem Biophys Res Commun"},{"key":"2022011223181945100_b8","doi-asserted-by":"crossref","first-page":"315","DOI":"10.3109\/08916939309079234","article-title":"Induction of TSH binding inhibitory immunoglobulins with the extracellular domain of human thyrotropin receptor produced using baculovirus expression system","volume":"14","author":"Seetharamaiah","year":"1993","journal-title":"Autoimmunity"},{"key":"2022011223181945100_b9","doi-asserted-by":"crossref","first-page":"103","DOI":"10.3109\/08916939409007983","article-title":"Induction of hyperthyroxinemia in BALB\/C but not in several other strains of mice","volume":"18","author":"Wagle","year":"1994","journal-title":"Autoimmunity"},{"key":"2022011223181945100_b10","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1046\/j.1365-2249.1998.00622.x","article-title":"Characterization of the murine immune response to the murine TSH receptor ectodomain. induction of hypothyroidism and TSH receptor antibodies","volume":"113","author":"Vlase","year":"1998","journal-title":"Clin Exp Immunol"},{"key":"2022011223181945100_b11","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1111\/j.1365-2249.1995.tb05548.x","article-title":"Unaltered thyroid function in mice responding to a highly immunogenic thyrotropin receptor. implications for the establishment of a mouse model for Graves\u2019 disease","volume":"99","author":"Carayanniotis","year":"1995","journal-title":"Clin Exp 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cells","volume":"126","author":"Yewdell","year":"1981","journal-title":"J Immunol"},{"key":"2022011223181945100_b39","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1146\/annurev.immunol.18.1.927","article-title":"DNA vaccines: immunology, application and optimization","volume":"18","author":"Gurunathan","year":"2000","journal-title":"Annu Rev Immunol"},{"key":"2022011223181945100_b40","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1084\/jem.189.1.169","article-title":"DNA vaccination: transfection and activation of dendritic cells as key events for immunity","volume":"189","author":"Akbari","year":"1999","journal-title":"J Exp Med"},{"key":"2022011223181945100_b41","doi-asserted-by":"crossref","first-page":"2388","DOI":"10.4049\/jimmunol.160.5.2388","article-title":"Contribution of cells at the site of DNA vaccination to the generation of antigen-specific immunity and memory","volume":"160","author":"Klinman","year":"1998","journal-title":"J 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