Name: preparation of mouse pituitary immunogen for the induction of experimental autoimmune hypophysitis
Uploaded: 2010-12-17T16:00:00
Description: Watch this Scientific Journal Video about Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis at JoVE.com

Detta arbete stöddes av NIH bidrag DK080351 till PC.

Experimentell Protokoll Det första steget i denna experimentella protokoll är att samla ett stort antal musen hypofys. Det andra steget är att homogenisera de körtlar att förbereda ett protein extrakt. Det tredje steget är att emulgera dessa proteiner med en fet blandning. Steg 1: Insamling och lagring av mus hypofys Musen hypofysen sitter vid basen av skallen i en depression av sphenoid ben kallas Sella turcica, omgiven sidled av t...

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	<li>Caturegli, P., Newschaffer, C., Olivi, A., Pomper, M. G., Burger, P. C., 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=.">.</a> Autoimmune Hypophysitis Endocr Rev. 26, 599-614 (2005).</li><li>Blansfield, J. A., Beck, K. E., Tran, K., Yang, J. C., Hughes, M. S., Kammula, U. S., Royal, R. E., Topalian, S. L., Haworth, L. R., Levy, C., Rosenberg, S. A., Sherry, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytotoxic+T-lymphocyte-associated+antigen-4+blockage+can+induce+autoimmune+hypophysitis+in+patients+with+metastatic+melanoma+and+renal+cancer.">Cytotoxic T-lymphocyte-associated antigen-4 blockage can induce autoimmune hypophysitis in patients with metastatic melanoma and renal cancer.</a> J Immunother. 28, 593-598 (2005).</li><li>Tzou, S. C., Lupi, I., Landek, M., Gutenberg, A., Tzou, Y. M., Kimura, H., Pinna, G., Rose, N. R., Caturegli, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoimmune+Hypophysitis+of+SJL+mice:+Clinical+Insights+from+a+New+Animal+Model.">Autoimmune Hypophysitis of SJL mice: Clinical Insights from a New Animal Model.</a> Endocrinology. 149, 3461-3469 (2008).</li><li>Goudie, R. B., Pinkerton, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anterior+hypophysitis+and+Hashimoto's+disease+in+a+woman.">Anterior hypophysitis and Hashimoto's disease in a woman.</a> J Pathol Bacteriol. 83, 584-585 (1962).</li><li>Caturegli, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Autoimmune+hypophysitis:+an+underestimated+disease+in+search+of+its+autoantigen(s).">Autoimmune hypophysitis: an underestimated disease in search of its autoantigen(s).</a> J Clin Endocrinol Metab. 92, 2038-2040 (2007).</li></ol>

Trots att den första patienten med autoimmuna hypophysitis rapporterades år 1962 fyra, patogena hypofysen autoantigener återstår att identifiera 5. Djurmodeller kan vara mycket användbara för att identifiera dessa autoantigener, medhjälp upptäckten av biomarkörer som är att överföra till patientvård. Denna artikel har visat ett enkelt förfarande för att förbereda hypofysen proteiner i en form som är tillräcklig för att inducera autoimmuna hypophysitis hos försöksdjur.

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<td>15-ml Falcon tubes</td>
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<td>homogenization buffer</td>
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<td>phosphate buffered saline</td>
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<td>protease inhibitor cocktail</td>
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<td>Polytron homogenizer</td>
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<td>BCA assay</td>
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<td>Pierce</td>
<td>23225</td>
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<td>2.5 mL Hamilton gastight syringe</td>
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<td>22-gauge micro-emulsifying needle</td>
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preparation of mouse pituitary immunogen for the induction of experimental autoimmune hypophysitis

Autoimmun hypophysitis är en kronisk inflammation i hypofysen orsakas eller åtföljas av autoimmunitet 1. Det har traditionellt ansetts vara en ovanlig sjukdom, men rapporteringen har ökat markant under senare år. Hypophysitis i själva verket utvecklas inte sällan som en &quot;bieffekt&quot; hos cancerpatienter som behandlas med antikroppar som blockerar inhibitoriska receptorer uttrycks på T-lymfocyter, som CTLA-4 2 och PD-1-receptorer. Autoimmuna hypophysitis kan induceras experimentellt genom att injicera möss med hypofysen proteiner blandat med ett adjuvans 3. I denna video artikeln visar vi hur du extraherar proteiner från mus hypofys och hur man förbereder dem i en form som lämpar sig för att inducera autoimmuna hypophysitis i SJL möss.

Watch this Scientific Journal Video about Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis at JoVE.com

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

Autoimmuna hypophysitis kan återges i möss genom att injicera ett utdrag ur mus hypofysen proteiner.

Beredning av Mouse Hypofys Immunogen för induktion av Experimentell Autoimmun Hypophysitis

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

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