Bu çalışma P01 A1078894 CAG Grant Ulusal Alerji Enstitüleri ve Enfeksiyon Hastalıkları tarafından desteklenen

<ol>
	<li>Nathan, C., Ding, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonresolving+Inflammation.">Nonresolving Inflammation.</a> Cell. 140 (6), 871-882 (2010).</li><li>Karin, M., Lawrence, T., Nizet, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Innate+immunity+gone+awry:+linking+microbial+infections+to+chronic+inflammation+and.">Innate immunity gone awry: linking microbial infections to chronic inflammation and.</a> 124, 823-835 (2006).</li><li>Connor, S. M., Taylor, C. E., Hughes, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emerging+infectious+determinants+of+chronic+diseases.">Emerging infectious determinants of chronic diseases.</a> Emerging Infectious Diseases. 12 (7), 1051-1057 (2006).</li><li>Barth, K., Remick, D. G., Genco, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+immune+regulation+by+microbial+pathogens+and+resulting+chronic+inflammation.">Disruption of immune regulation by microbial pathogens and resulting chronic inflammation.</a> Journal of Cellular Physiology. , (2012).</li><li>Hayashi, C., Gudino, C. V., Gibson, F. C., Genco, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review:+Pathogen-induced+inflammation+at+sites+distant+from+oral+infection:+bacterial+persistence+and+induction+of+cell-specific+innate+immune+inflammatory+pathways.">Review: Pathogen-induced inflammation at sites distant from oral infection: bacterial persistence and induction of cell-specific innate immune inflammatory pathways.</a> Molecular Oral Microbiology. 25 (5), 305-316 (2010).</li><li>Gibson, F. C., Ukai, T., Genco, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engagement+of+specific+innate+immune+signaling+pathways+during+Porphyromonas+gingivalis+induced+chronic+inflammation+and+atherosclerosis.">Engagement of specific innate immune signaling pathways during Porphyromonas gingivalis induced chronic inflammation and atherosclerosis.</a> Frontiers in Bioscience: a Journal and Virtual Library. 13, 2041-2059 (2008).</li><li>Pihlstrom, B. L., Michalowicz, B. S., Johnson, N. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Periodontal+diseases.">Periodontal diseases.</a> Lancet. 366 (9499), 1809-1820 (2005).</li><li>Baker, P. J., Evans, R. T., Roopenian, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+infection+with+Porphyromonas+gingivalis+and+induced+alveolar+bone+loss+in+immunocompetent+and+severe+combined+immunodeficient+mice.">Oral infection with Porphyromonas gingivalis and induced alveolar bone loss in immunocompetent and severe combined immunodeficient mice.</a> Archives of Oral Biology. 39 (12), 1035-1040 (1994).</li><li>Baker, P. J., Carter, S., Dixon, M., Evans, R. T., Roopenian, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serum+antibody+response+to+oral+infection+precedes+but+does+not+prevent+Porphyromonas+gingivalis-induced+alveolar+bone+loss+in+mice.">Serum antibody response to oral infection precedes but does not prevent Porphyromonas gingivalis-induced alveolar bone loss in mice.</a> Oral Microbiology and Immunology. 14 (3), 194-196 (1999).</li><li>Gibson, F. C., Hong, 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=Innate+immune+recognition+of+invasive+bacteria+accelerates+atherosclerosis+in+apolipoprotein+E-deficient+mice.">Innate immune recognition of invasive bacteria accelerates atherosclerosis in apolipoprotein E-deficient mice.</a> Circulation. 109 (22), 2801-2806 (2004).</li><li>Hayashi, C., Viereck, 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=Porphyromonas+gingivalis+accelerates+inflammatory+atherosclerosis+in+the+innominate+artery+of+ApoE+deficient+mice.">Porphyromonas gingivalis accelerates inflammatory atherosclerosis in the innominate artery of ApoE deficient mice.</a> Atherosclerosis. 215 (1), 52-59 (2011).</li><li>Hayashi, C., Madrigal, A. G., 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=Pathogen-mediated+inflammatory+atherosclerosis+is+mediated+in+part+via+Toll-like+receptor+2-induced+inflammatory+responses.">Pathogen-mediated inflammatory atherosclerosis is mediated in part via Toll-like receptor 2-induced inflammatory responses.</a> Journal of Innate Immunity. 2 (4), 334-343 (2010).</li><li>Hayashi, C., Papadopoulos, G., 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=Protective+role+for+TLR4+signaling+in+atherosclerosis+progression+as+revealed+by+infection+with+a+common+oral+pathogen.">Protective role for TLR4 signaling in atherosclerosis progression as revealed by infection with a common oral pathogen.</a> Journal of Immunology (Baltimore, Md.: 1950). 189 (7), 3681-3688 (2012).</li><li>Papadopoulos, G., Weinberg, E. O., 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=Macrophage-Specific+TLR2+Signaling+Mediates+Pathogen-Induced+TNF-Dependent+Inflammatory+Oral+Bone+Loss.">Macrophage-Specific TLR2 Signaling Mediates Pathogen-Induced TNF-Dependent Inflammatory Oral Bone Loss.</a> The Journal of Immunology. , (2012).</li><li>Gibson, F. C., Hong , 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=Innate+immune+recognition+of+invasive+bacteria+accelerates+atherosclerosis+in+apolipoprotein+E-deficient+mice.">Innate immune recognition of invasive bacteria accelerates atherosclerosis in apolipoprotein E-deficient mice.</a> Circulation. 109 (22), 2801-2806 (2004).</li><li>Miyamoto, T., Yumoto, H., Takahashi, Y., Davey, M., Gibson, F. C., Genco, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogen-accelerated+atherosclerosis+occurs+early+after+exposure+and+can+be+prevented+via+immunization.">Pathogen-accelerated atherosclerosis occurs early after exposure and can be prevented via immunization.</a> Infection and Immunity. 74 (2), 1376-1380 (2006).</li><li>Baker, P. J., Dixon, M., Roopenian, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+control+of+susceptibility+to+Porphyromonas+gingivalis-induced+alveolar+bone+loss+in+mice.">Genetic control of susceptibility to Porphyromonas gingivalis-induced alveolar bone loss in mice.</a> Infection and Immunity. 68 (10), 5864-5868 (2000).</li><li>Baker, P. J., Dixon, M., Evans, R. T., Roopenian, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+of+Porphyromonas+gingivalis+strains+in+the+induction+of+alveolar+bone+loss+in+mice.">Heterogeneity of Porphyromonas gingivalis strains in the induction of alveolar bone loss in mice.</a> Oral Microbiology and Immunology. 15 (1), 27-32 (2000).</li><li>Daugherty, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+atherosclerosis.">Mouse models of atherosclerosis.</a> The American Journal of the Medical Sciences. 323 (1), 3-10 (2002).</li><li>Weinreb, D. B., Aguinaldo, J. G. S., Feig, J. E., Fisher, E. A., Fayad, Z. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+MRI+of+mouse+models+of+atherosclerosis.">Non-invasive MRI of mouse models of atherosclerosis.</a> NMR in Biomedicine. 20 (3), 256-264 (2007).</li></ol>

Yazarlar hiçbir rakip mali çıkarlarının olmadığını beyan ederim.

S. gingivalis ağızdan enfeksiyon modeli lokal ve sistemik siteleri patojen kaynaklı kronik enflamasyon çalışma için değerli bir araç sağlar. Bu benzersiz model, kronik inflamasyon ve İmmünopatolojiye katkıda hem ev sahibi ve patojen spesifik mekanizmaların karakterizasyonu izin verir. Buna ek olarak, bağışıklık kazandırma ve model farmakolojik müdahale dahil, yeni tedavi stratejileri, taranması için de kullanılabilir. Bu protokolde belirtilen adımlar P. başlamasını, ilerleme...

Kronik inflamasyon patolojik doku hasarı ve insanlarda birçok kronik hastalıkların bir birleştirici özelliği önemli bir sürücüdür. Bu hastalıklar neoplastik, otoimmün ve kronik enflamatuar hastalıklar 1 içerir. Birçok kronik hastalığın etyolojisi bilinmemektedir ama genetik yatkınlık ve çevresel faktörlerin de tanışmasını içeren, karmaşık ve çok faktörlü olduğu anlaşılmaktadır. Enflamasyon ettiricileri zor olmaya devam ederken immün aktivasyon hücresel ve moleküler profilleri patojenlere karşı ev sahibi tepkilerin 2 gözlenen desenleri ile önemli ölçüde üst üste gelir. Montaj kanıt mikrobiyal gelişme patojenler ve kronik inflamasyon ilerlemesi ve onun çeşitli klinik belirtiler 2,3 ile enfeksiyon implicates. Patojenler neden ve kalıcı enfeksiyonlar konak bağışıklık sistemini yıkarak ve kurarak kronik enflamasyonu doğrudan sürdürmek <sup&gt; 4. Mikrobiyal kalıcılık yokluğunda, enfeksiyon, self-antijenlere moleküler taklidin tetiklenen oto-bağışıklık tepkilerinin, imünojenik hale getirmek, self-antijenlere ya da daha önce maskeli ana antijenleri açıklamaları zarar değişikliklerden kaynaklanan kronik enflamasyonu hızlandırabilir. Nadiren, ancak spesifik patojenler özel bir kronik hastalık evrensel nedeni olarak tanımlanmıştır. Aksine, mevcut verilerin çoğunluğu patojenler genetik olarak yatkın konak 3 klinik belirtileri ve hastalık sonuçları geniş bir yelpazede ile kronik inflamasyon ortaya çıkarmak için farklı mekanizmaları kullandığını iddia. Böylece, spesifik patojenler önemli bir halk sağlığı etkileri yanı sıra, pek çok kronik hastalıkların tedavisi ve önlenmesi olabilir kronik enflamasyona neden olan mekanizmaların ayrıntılı bir anlayış. Kronik inflamasyon indüksiyon ve bakım katkıda konak ve patojen spesifik mekanizmalar olmasına rağmenkötü patojen kaynaklı kronik inflamasyon modellemede gelişmeler bu süreçlerin anlayışımızı ilerletmek için başladı, anladım. S. gingivalis oral enfeksiyon modeli konak ve patojen lokal (oral kemik kaybı) kronik enflamasyon katkıda özgü mekanizmaları ve sistemik siteleri (ateroskleroz) 5,6 analizi izin verir patojen kaynaklı kronik inflamasyon eşsiz, iyi karakterize fare modelidir. S. gingivalis insan periodontal hastalık, doku 7 nolu destekleme dişin tahribatı ile karakterize edilen bir enfeksiyon kaynaklı kronik enflamatuar hastalıkta rol oynayan bir gram-negatif anaerob ağızdan patojendir. Enfeksiyonun başlangıç ​​yerinde patoloji ek olarak, artan kanıtlar P. implicates gingivalis aterosklerozun gelişiminde ve 5 dahil sistemik hastalıkların ilerlemesinde kronik inflamasyon -sebepli, bir hastalık kronik iltihabın özelliği, arter damar duvarının: n. P. olan spesifik patojensiz farelerde ağızdan enfeksiyonu gingivalis alveoler kemik 8 destek dişin tahribatıyla sonuçlanan bir inflamatuar yanıtı tetikler. P. gingivalis 42 gün Enfeksiyondan 8 ve farenin patogene-özgü serum antikor titrelerinin yüksek seviyelerini 9 geliştirmek için, enfekte edilmiş farelerin ağızlarından elde edilebilir. - / - Apo-E kullanılarak aterosklerozun yerleşmiş fare modelinde farenin (Apo - / -), P. ile ağızdan enfeksiyondan gingivalis aort sinüs 10 ve innominat arter 11 inflamatuar plak birikimini sürücüler kronik enflamasyona neden olur. P. innominat arter içinde ilerici inflamasyon gingivalis farenin in vivo MRG kullanılarak canlı hayvanlarda izlenebilir ğında. Histolojik olarak, P., arter lezyonlar gingivalis farenin lipidler accompa artmış birikimini sergilemek ğındavasküler endotel aktivasyonu, artmış bir immün hücre infiltrasyonu ve iltihap aracılarının 12 artmış ekspresyonu ile reddedilmiş. Nakavt farelerde, bu modelin kullanımı, ana sinyal parçalan ve iltihaplanma aracılan rolü, hem de hücre S. sürücü spesifik etkileşimler açıklamıştır 14 - gingivalis immünopatoloji 12 -kaynaklı. Buna ek olarak, tarif bakteriyel mutantlar kullanılarak deneyler önemli S. belirledik gingivalis lokal ve sistemik sitelerinde 15 kronik enflamasyon katkıda bulunan faktörleri virülansa. Bu makale P. değerlendirilmesi için yöntemler detayları gingivalis lokal ve sistemik sitelerinde kronik inflamasyon -kaynaklı. Bu Witt yazılımı kullanılarak mikroBT alveol kemiği kaybı analizi için ayrıntılı bir protokol sağlar. Ayrıca, biz ilerici değerlendirmesinde vivo, canlı hayvan MRG serial yararını tanımlamakinnominat arter içinde inflamasyon. Biz görselleştirme ve arteriyel lezyonlarda inflamatuar plak ölçümü için yöntemler içerir ve histolojik karakterizasyonu açıklar. Transgenik farelerden ve tanımlı bakteriyel mutantlarının kullanımı ana ve hastalığın başlaması, ilerlemesi ve sonuç olarak ilgili mikrobiyal faktörlerini tanımlamak için bu model özellikle uygun hale getirir. Buna ek olarak, model, aşı ve farmakolojik müdahale dahil, yeni tedavi stratejileri, taranması için de kullanılabilir.

<table border="0" cellpadding="0" cellspacing="0" width="924">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:508px;">Name of the Material/Eqiupment</td>
			<td style="width:196px;">Company</td>
			<td style="width:132px;">Catalogue Number</td>
			<td style="width:88px;">Comments&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Amira analysis software</td>
			<td>Visualization Sciences Group</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anaerobic chamber DW Scientific Model MG500&#160;</td>
			<td>Microbiology International</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">BHI&#160;</td>
			<td>Becton-Dickinson&#160;</td>
			<td>211059</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hemin&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>51280-5G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Menadione (Vitamin K)</td>
			<td>Sigma-Aldrich&#160;&#160;</td>
			<td>M5625-25G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Yeast Extract</td>
			<td>Becton-Dickinson&#160;</td>
			<td>212750</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Carboxymethyl cellulose (medium viscocity)&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>C-4888</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sulfamethoxazole and Trimethoprim Oral Suspension 200 mg/40 mg per 5ml</td>
			<td>Hi-Tech Pharmacal</td>
			<td>NDC 50383-823-16</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#956;CT 40&#160;</td>
			<td>Scanco</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HistoChoice Tissue Fixative</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>H2904</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sudan IV</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>S4261-25G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">vertical-bore 11.7T Avance spectrometer&#160;</td>
			<td>Bruker</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paravision</td>
			<td>Paravision</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ImageJ</td>
			<td>NIH</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">rat anti-mouse F4/80</td>
			<td>Serotec</td>
			<td>MCA497R</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">rat anti-mouse TLR2</td>
			<td>eBioscience</td>
			<td>13-9021-80</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Leica S4 Dissecting Scope</td>
			<td>Leica</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microm HM 550 Cryostat</td>
			<td>Microm</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

a mouse model for pathogen-induced chronic inflammation at local and systemic sites

Kronik inflamasyon patolojik doku hasarı ve neoplastik, otoimmün ve kronik iltihaplı hastalıklar, insanlarda pek çok kronik hastalıkların bir birleştirici özelliği önemli bir sürücüdür. Delil gelişen klinik belirtilerin çok çeşitli geliştirme ve kronik hastalıkların ilerlemesinde patojen kaynaklı kronik inflamasyon implicates. Kronik hastalığı karmaşık ve çok faktörlü etiyolojisi, nedensellik kanıtı ve mekanik bağlantıların kurulması için deney tasarlama insanlarda neredeyse imkansız. Hayvan modelleri kullanmanın bir avantajı özel bir hastalığın seyrini etkilediği, her ikisi de, genetik ve çevresel faktörler kontrol edilebilir olmasıdır. Böylece, enfeksiyon ilgili hayvan modelleri tasarlarken kronik inflamasyona katkıda konak ve patojen spesifik mekanizmaları belirlemede önemli bir adımı temsil etmektedir. Burada patojen kaynaklı kronik Serma bir fare modeli tarifOral patojen Porphyromonas gingivalis, çok insanlarda periodontal hastalık ile bağlantılı bir bakteri ile enfeksiyondan sonra, lokal ve sistemik sitelerde mmation. Özgül patojen içermeyen farelere ağızdan enfeksiyonu alveoler kemik, periodontal hastalık bir işaretidir destek dişin tahribatı ile sonuçlanan bir inflamatuar yanıtı tetikler. P. ile kurulmuş bir fare ateroskleroz modelinde, enfeksiyon gingivalis, vasküler endotel aktivasyonu, artmış bir immün hücre infiltrasyonu ve lezyonlarında iltihap aracılarının yükselen sentezlenmesi ile birlikte aort sinüs ve iliyak arteri içindeki inflamatuar plak birikimini hızlandırır. Biz lokal ve sistemik sitelerinde inflamasyon değerlendirilmesi için ayrıntılı metodolojiler. Transgenik farelerden ve tanımlı bakteriyel mutantlarının kullanımı ana ve hastalığın başlaması, ilerlemesi ve sonuç olarak ilgili mikrobiyal faktörlerini tanımlamak için bu model özellikle uygun hale getirir. Additionally modeli aşı ve farmakolojik müdahale dahil, yeni tedavi stratejileri, taranması için de kullanılabilir.

Uygun fare modeli ve oral enfeksiyon rejimi, S. kullanma gingivalis kronik inflamasyon ve Immunopathology yerel (oral kavite) seviyesinde ve sistemik the (atardamar) (Şekil 1) indükler. Farelerde, P. ile ağızdan enfeksiyondan gingivalis alveoler kemik 8 destek dişin yok edilmesini sağlayan bir inflamatuar yanıtı tetikler. P. gingivalis farenin ağırlıklı IgG izo 9 vardır, bu organizma için, serum...

Watch this Scientific Journal Video about A Mouse Model for Pathogen-induced Chronic Inflammation at Local and Systemic Sites at JoVE.com

A Mouse Model for Pathogen-induced Chronic Inflammation at Local and Systemic Sites

Hayvan modelleri kronik inflamasyon gelişimine katkıda konak ve patojen spesifik mekanizmaları tanımlarken paha biçilmez araçlardır olduğu kanıtlanmıştır. Burada, lokal ve sistemik inflamasyon bölgelerinde ilerlemesini değerlendirmek için insan patojeni Porphyromonas gingivalis ve detay metodolojileri ile ağızdan enfeksiyondan bir fare modeli tarif eder.

Lokal ve sistemik Yer de patojen kaynaklı kronik enflamasyon Fare Modeli

Chronic inflammation is a major driver of pathological tissue damage and a unifying characteristic of many chronic diseases in humans including ...

a-mouse-model-for-pathogen-induced-chronic-inflammation-at-local

Research

JoVE Journal

Immunology and Infection

17.4K Görüntüleme.  Boston University School of Medicine.  Hayvan modelleri kronik inflamasyon gelişimine katkıda konak ve patojen spesifik mekanizmaları tanımlarken paha biçilmez araçlardır olduğu kanıtlanmıştır. Burada, lokal ve sistemik inflamasyon bölgelerinde ilerlemesini değerlendirmek için insan patojeni Porphyromonas gingivalis ve detay metodolojileri ile ağızdan enfeksiyondan bir fare modeli tarif eder. 

Video: A Mouse Model for Pathogen-induced Chronic Inflammation at Local and Systemic Sites

Trichuris muris Infection: A Model of Type 2 Immunity and Inflammation in the Gut

Trichuris muris is a natural pathogen of mice and is biologically and antigenically similar to species of Trichuris that 
infect humans and livestock1. Infective eggs are given by oral gavage, hatch in the distal small intestine, invade the intestinal epithelial cells (IECs) 
that line the crypts of the cecum and proximal colon and upon maturation the worms release eggs into the environment1. This model is a powerful tool to 
examine factors that control CD4+ T helper (Th) cell activation as well as changes in the intestinal epithelium. The immune response that occurs in 
resistant inbred strains, such as C57BL/6 and BALB/c, is characterized by Th2 polarized cytokines (IL-4, IL-5 and IL-13) and expulsion of worms while Th1-associated 
cytokines (IL-12, IL-18, IFN-&gamma;) promote chronic infections in genetically susceptible AKR/J mice2-6. Th2 cytokines promote physiological changes in 
the intestinal microenvironment including rapid turnover of IECs, goblet cell differentiation, recruitment and changes in epithelial permeability and smooth muscle 
contraction, all of which have been implicated in worm expulsion7-15. Here we detail a protocol for propagating Trichuris muris eggs which can 
be used in subsequent experiments. We also provide a sample experimental harvest with suggestions for post-infection analysis. Overall, this protocol will provide 
researchers with the basic tools to perform a Trichuris muris mouse infection model which can be used to address questions pertaining to Th proclivity in 
the gastrointestinal tract as well as immune effector functions of IECs.

Trichuris muris Infection: A Model of Type 2 Immunity and Inflammation in the Gut

Trichuris muris infection is an intestinal model of Th2 immunity where resistant mice generate a protective Th2 response and susceptible mice generate a pathological Th1 response.

Trichuris muris Enfeksiyon: Gut Tip 2 Bağışıklık ve İnflamasyon Bir Model

Trichuris muris is a natural pathogen of mice and is biologically and antigenically similar to species of Trichuris that 
infect humans and livestock1. ...

İmmünoloji ve Enfeksiyon

Using the BLT Humanized Mouse as a Stem Cell based Gene Therapy Tumor Model

Small animal models such as mice have been extensively used to study human disease and to develop new therapeutic interventions. Despite the wealth of information gained from these studies, the unique characteristics of mouse immunity as well as the species specificity of viral diseases such as human immunodeficiency virus (HIV) infection led to the development of humanized mouse models. The earlier models involved the use of C. B 17 scid/scid mice and the transplantation of human fetal thymus and fetal liver termed thy/liv (SCID-hu) 1, 2 or the adoptive transfer of human peripheral blood leukocytes (SCID-huPBL) 3. Both models were mainly utilized for the study of HIV infection.
One of the main limitations of both of these models was the lack of stable reconstitution of human immune cells in the periphery to make them a more physiologically relevant model to study HIV disease. To this end, the BLT humanized mouse model was developed. BLT stands for bone marrow/liver/thymus. In this model, 6 to 8 week old NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) immunocompromised mice receive the thy/liv implant as in the SCID-hu mouse model only to be followed by a second human hematopoietic stem cell transplant 4. The advantage of this system is the full reconstitution of the human immune system in the periphery. This model has been used to study HIV infection and latency 5-8.
We have generated a modified version of this model in which we use genetically modified human hematopoietic stem cells (hHSC) to construct the thy/liv implant followed by injection of transduced autologous hHSC 7, 9. This approach results in the generation of genetically modified lineages. More importantly, we adapted this system to examine the potential of generating functional cytotoxic T cells (CTL) expressing a melanoma specific T cell receptor. Using this model we were able to assess the functionality of our transgenic CTL utilizing live positron emission tomography (PET) imaging to determine tumor regression (9).
The goal of this protocol is to describe the process of generating these transgenic mice and assessing in vivo efficacy using live PET imaging. As a note, since we use human tissues and lentiviral vectors, our facilities conform to CDC NIH guidelines for Biosafety Level 2 (BSL2) with special precautions (BSL2+). In addition, the NSG mice are severely immunocompromised thus, their housing and maintenance must conform to the highest health standards (<a href="http://jaxmice.jax.org/research/immunology/005557-housing.html" target="_blank">http://jaxmice.jax.org/research/immunology/005557-housing.html</a>).

The generation and characterization of tumor specific T cells using humanized mice is described here. Human thymic tissue and genetically modified human hematopoietic stem cells are transplanted into immunocompromised mice. This results in the reconstitution of an engineered human immune system allowing for in vivo examination of anti-tumor immune responses.

Bir Kök Hücre tabanlı Gen Terapisi Tümör Modeli olarak BLT Insanlaşmış Fare Kullanma

Small animal models such as mice have been extensively used to study human  disease and to develop new therapeutic interventions. Despite the wealth of  ...

In vivo Imaging Method to Distinguish Acute and Chronic Inflammation

Inflammation is a fundamental aspect of many human diseases. In this video report, we demonstrate non-invasive bioluminescence imaging techniques that distinguish acute and chronic inflammation in mouse models. With tissue damage or pathogen invasion, neutrophils are the first line of defense, playing a major role in mediating the acute inflammatory response. As the inflammatory reaction progresses, circulating monocytes gradually migrate into the site of injury and differentiate into mature macrophages, which mediate chronic inflammation and promote tissue repair by removing tissue debris and producing anti-inflammatory cytokines. Intraperitoneal injection of luminol (5-amino-2,3-dihydro-1,4-phthalazinedione, sodium salt) enables detection of acute inflammation largely mediated by tissue-infiltrating neutrophils. Luminol specifically reacts with the superoxide generated within the phagosomes of neutrophils since bioluminescence results from a myeloperoxidase (MPO) mediated reaction. Lucigenin (bis-N-methylacridinium nitrate) also reacts with superoxide in order to generate bioluminescence. However, lucigenin bioluminescence is independent of MPO and it solely relies on phagocyte NADPH oxidase (Phox) in macrophages during chronic inflammation. Together, luminol and lucigenin allow non-invasive visualization and longitudinal assessment of different phagocyte populations across both acute and chronic inflammatory phases. Given the important role of inflammation in a variety of human diseases, we believe this non-invasive imaging method can help investigate the differential roles of neutrophils and macrophages in a variety of pathological conditions.

In vivo Imaging Method to Distinguish Acute and Chronic Inflammation

We describe a non-invasive imaging method for distinguishing inflammatory stages. Systemic delivery of luminol reveals areas of acute inflammation dependent upon MPO activity in neutrophils. In contrast, injection of lucigenin allows for visualization of chronic inflammation dependent upon Phox activity in macrophages.

In vivo Görüntüleme Yöntemi Akut ve Kronik İltihap ayırt

Inflammation is a fundamental aspect of many human diseases. In this video report, we demonstrate non-invasive bioluminescence imaging techniques that ...

In vitro Coculture Assay to Assess Pathogen Induced Neutrophil Trans-epithelial Migration

Mucosal surfaces serve as protective barriers against pathogenic organisms.&#160;Innate immune responses are activated upon sensing pathogen leading to the infiltration of tissues with migrating inflammatory cells, primarily neutrophils.&#160;This process has the potential to be destructive to tissues if excessive or held in an unresolved state.&#160; Cocultured in vitro models can be utilized to study the unique molecular mechanisms involved in pathogen induced neutrophil trans-epithelial migration.&#160;This type of model provides versatility in experimental design with opportunity for controlled manipulation of the pathogen, epithelial barrier, or neutrophil.&#160;Pathogenic infection of the apical surface of polarized epithelial monolayers grown on permeable transwell filters instigates physiologically relevant basolateral to apical trans-epithelial migration of neutrophils applied to the basolateral surface.&#160;The in vitro model described herein demonstrates the multiple steps necessary for demonstrating neutrophil migration across a polarized lung epithelial monolayer that has been infected with pathogenic P. aeruginosa (PAO1).&#160;Seeding and culturing of permeable transwells with human derived lung epithelial cells is described, along with isolation of neutrophils from whole human blood and culturing of PAO1 and nonpathogenic K12 E. coli (MC1000).&#160; The emigrational process and quantitative analysis of successfully migrated neutrophils that have been mobilized in response to pathogenic infection is shown with representative data, including positive and negative controls.&#160;This in vitro model system can be manipulated and applied to other mucosal surfaces.&#160;Inflammatory responses that involve excessive neutrophil infiltration can be destructive to host tissues and can occur in the absence of pathogenic infections.&#160;A better understanding of the molecular mechanisms that promote neutrophil trans-epithelial migration through experimental manipulation of the in vitro coculture assay system described herein has significant potential to identify novel therapeutic targets for a range of mucosal infectious as well as inflammatory diseases.

In vitro Coculture Assay to Assess Pathogen Induced Neutrophil Trans-epithelial Migration

Neutrophil trans-epithelial migration in response to mucosal bacterial infection contributes to epithelial injury and clinical disease.&#160;An in vitro model has been developed that combines pathogen, human neutrophils, and polarized human epithelial cell layers grown on transwell filters to facilitate investigations towards unraveling the molecular mechanisms orchestrating this phenomenon.

Tüp bebek Patojen Kaynaklı Nötrofil Trans-epitel Göçünü Değerlendirmek için Coculture Tahlili

Mucosal surfaces serve as protective barriers against pathogenic organisms.&#160;Innate immune responses are activated upon sensing pathogen leading to ...

Systemic Injection of Neural Stem/Progenitor Cells in Mice with Chronic EAE

Neural stem/precursor cells (NPCs) are a promising stem cell source for transplantation approaches aiming at brain repair or restoration in regenerative neurology. This directive has arisen from the extensive evidence that brain repair is achieved after focal or systemic NPC transplantation in several preclinical models of neurological diseases.
These experimental data have identified the cell delivery route as one of the main hurdles of restorative stem cell therapies for brain diseases that requires urgent assessment. Intraparenchymal stem cell grafting represents a logical approach to those pathologies characterized by isolated and accessible brain lesions such as spinal cord injuries and Parkinson's disease. Unfortunately, this principle is poorly applicable to conditions characterized by a multifocal, inflammatory and disseminated (both in time and space) nature, including multiple sclerosis (MS). As such, brain targeting by systemic NPC delivery has become a low invasive and therapeutically efficacious protocol to deliver cells to the brain and spinal cord of rodents and nonhuman primates affected by experimental chronic inflammatory damage of the central nervous system (CNS).
This alternative method of cell delivery relies on the NPC pathotropism, specifically their innate capacity to (i) sense the environment via functional cell adhesion molecules and inflammatory cytokine and chemokine receptors; (ii) cross the leaking anatomical barriers after intravenous (i.v.) or intracerebroventricular (i.c.v.) injection; (iii) accumulate at the level of multiple perivascular site(s) of inflammatory brain and spinal cord damage; and (i.v.) exert remarkable tissue trophic and immune regulatory effects onto different host target cells in vivo.
Here we describe the methods that we have developed for the i.v. and i.c.v. delivery of syngeneic NPCs in mice with experimental autoimmune encephalomyelitis (EAE), as model of chronic CNS inflammatory demyelination, and envisage the systemic stem cell delivery as a valuable technique for the selective targeting of the inflamed brain in regenerative neurology.

The transplantation of neural stem/progenitor cells (NPCs) holds great promises in regenerative neurology. The systemic delivery of NPCs has turned into effective, low invasive, and therapeutically very efficacious protocol to deliver stem cells in the brain and spinal cord of rodents and nonhuman primates affected by experimental chronic inflammatory damage of the central nervous system.

Kronik EAE ile Fare nöral kök / progenitör hücrelerinin sistemik enjeksiyonundan

Neural stem/precursor cells (NPCs) are a promising stem cell source for transplantation approaches aiming at brain repair or restoration in regenerative ...

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages and disadvantages. We describe here an experimental colitis model that is initiated by adoptive transfer of syngeneic splenic CD4+CD45RBhigh T cells into T and B cell deficient recipient mice. The CD4+CD45RBhigh T cell population that largely consists of na&#239;ve effector cells is capable of inducing chronic intestinal inflammation, closely resembling key aspects of human IBD. This method can be manipulated to study aspects of disease onset and progression. Additionally it can be used to study the function of innate, adaptive, and regulatory immune cell populations, and the role of environmental exposures, i.e., the microbiota, in intestinal inflammation. In this article we illustrate the methodology for inducing colitis with a step-by-step protocol. This includes a video demonstration of key technical aspects required to successfully develop this murine model of experimental colitis for research purposes.

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Here, we present a protocol to induce colonic inflammation in mice by adoptive transfer of syngeneic CD4+CD45RBhigh T cells into T and B cell deficient recipients. Clinical and histopathological features mimic human inflammatory bowel diseases. This method allows the study of the initiation of colonic inflammation and progression of disease.

Efektör CD4 Evlatlık transferi ile Kemirgen Bağırsak Enflamasyon indüksiyonu<sup&gt; +</sup&gt; CD45RB<sup&gt; Yüksek</sup&gt; Bağışık yetmezliği olan farelere içine T Hücreleri

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages ...

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Studying the interaction between proteins is key in understanding their function(s). A very powerful method that is frequently used to study interactions of proteins with other macromolecules in a complex sample is called co-immunoprecipitation. The described co-immunoprecipitation protocol allows to demonstrate and further investigate the interaction between the antiviral myxovirus resistance protein 1 (Mx1) and one of its viral targets, the influenza A virus nucleoprotein (NP). The protocol starts with transfected mammalian cells, but it is also possible to use influenza A virus infected cells as starting material. After cell lysis, the viral NP protein is pulled-down with a specific antibody and the resulting immune-complexes are precipitated with protein G beads. The successful pull-down of NP and the co-immunoprecipitation of the antiviral Mx1 protein are subsequently revealed by western blotting. A prerequisite for successful co-immunoprecipitation of Mx1 with NP is the presence of N-ethylmaleimide (NEM) in the cell lysis buffer. NEM alkylates free thiol groups. Presumably this reaction stabilizes the weak and/or transient NP&#8211;Mx1 interaction by preserving a specific conformation of Mx1, its viral target or an unknown third component. An important limitation of co-immunoprecipitation experiments is the inadvertent pull-down of contaminating proteins, caused by nonspecific binding of proteins to the protein G beads or antibodies. Therefore, it is very important to include control settings to exclude false positive results. The described co-immunoprecipitation protocol can be used to study the interaction of Mx proteins from different vertebrate species with viral proteins, any pair of proteins, or of a protein with other macromolecules. The beneficial role of NEM to stabilize weak and/or transient interactions needs to be tested for each interaction pair individually.

This co-immunoprecipitation protocol allows to study the interaction between the influenza A virus nucleoprotein and the antiviral Mx1 protein in human cells. The protocol emphasizes the importance of N-ethylmaleimide for successful co-immunoprecipitation of Mx1 and influenza A virus nucleoprotein.

İnfluenza A virüsü nükleoproteini Fare MX1 Protein Co-immunopresipitasyonu

Studying the interaction between proteins is key in understanding their function(s). A very powerful method that is frequently used to study interactions ...

An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen

The ability to rapidly secrete cytokines upon stimulation is a functional characteristic of the invariant natural killer T (iNKT) cell lineage. iNKT cells are therefore characterized as an innate T cell population capable of activating and steering adaptive immune responses. The development of improved techniques for the culture and expansion of murine iNKT cells facilitates the study of iNKT cell biology in in vitro and in vivo model systems. Here we describe an optimized procedure for the isolation and expansion of murine splenic iNKT cells.
Spleens from C57Bl/6 mice are removed, dissected and strained and the resulting cellular suspension is layered over density gradient media. Following centrifugation, splenic mononuclear cells (MNCs) are collected and CD5-positive (CD5+) lymphocytes are enriched for using magnetic beads. iNKT cells within the CD5+ fraction are subsequently stained with &#945;GalCer-loaded CD1d tetramer and purified by fluorescence activated cell sorting (FACS). FACS sorted iNKT cells are then initially cultured in vitro using a combination of recombinant murine cytokines and plate-bound T cell receptor (TCR) stimuli before being expanded in the presence of murine recombinant IL-7. Using this technique, approximately 108 iNKT cells can be generated within 18-20 days of culture, after which they can be used for functional assays in vitro, or for in vivo transfer experiments in mice.

Here we present an adapted protocol that can be used to generate a large number of murine invariant natural killer T cells from mouse spleen. The protocol outlines an approach by which splenic iNKT cells can be enriched for, isolated and expanded in vitro using a limited number of animals and reagents.

Fare Dalak Değişmeyen Doğal Öldürücü T Hücreleri İzolasyon ve Yaygınlaştırılması için optimize Yöntemi

The ability to rapidly secrete cytokines upon stimulation is a functional characteristic of the invariant natural killer T (iNKT) cell lineage. iNKT cells ...

Intra-tracheal Administration of Haemophilus influenzae in Mouse Models to Study Airway Inflammation

Here, we describe a detailed procedure to efficiently and directly deliver Haemophilus influenzae into the lower respiratory tracts of mice. We demonstrate the procedure for preparing H. influenzae inoculum, intra-tracheal instillation of H. influenzae into the lung, collection of broncho-alveolar lavage fluid (BALF), analysis of immune cells in the BALF, and RNA isolation for differential gene expression analysis. This procedure can be used to study the lung inflammatory response to any bacteria, virus or fungi. Direct tracheal instillation is mostly preferred over intranasal or aerosol inhalation procedures because it more efficiently delivers the bacterial inoculum into the lower respiratory tract with less ambiguity.

Intra-tracheal Administration of Haemophilus influenzae in Mouse Models to Study Airway Inflammation

We demonstrate the procedure for intra-tracheal inoculation of Haemophilus influenzae into the lower respiratory tracts of mice. This is a very useful tool to study signaling pathways that regulate airway inflammation in mouse models.

intra-trakeal İdaresi<em&gt; Haemophilus influenza</em&gt; Fare Modelleri Havayolu inflamasyonu Eğitim için

Here, we describe a detailed procedure to efficiently and directly deliver Haemophilus influenzae into the lower respiratory tracts of mice. We ...

Mouse Body Temperature Measurement Using Infrared Thermometer During Passive Systemic Anaphylaxis and Food Allergy Evaluation

Mouse body temperature measurement is of paramount importance for investigating allergies and anaphylactic symptoms. Rectal probes for temperature readings is common, and they have been proven to be accurate and invaluable in this regard. However, this method of temperature measurement requires the mice to be anesthetized in order to insert the probe without injury to the animal. This limits the ability to observe other phenotypes of the mouse simultaneously. In order to investigate other phenotypes while measuring temperatures, rectal probes are not ideal, and another method is desired. Here, we introduce a noninvasive method of temperature measurement that foregoes the requirement for mouse anesthesia while maintaining equal reliability to rectal probes in measuring body temperature. We use an infrared thermometer that detects body surface temperatures at ranges between 2 and 150 mm. This method of body temperature measurement is successful in reliably replicating temperature change trends during passive system anaphylaxis experiments in mice. We show that body surface temperatures are about 2.0 &#176;C lower than rectal probe measurements, but the degree of temperature drop follows the same trend. Furthermore, we use the same technique to observe mice in a food allergy model to evaluate temperature and activity levels simultaneously.

Here we present a new method to accurately measure body temperature differences in passive systemic anaphylaxis (PSA) and food allergy mouse models using an infrared thermometer. This procedure has been accurately duplicated in previous PSA results.