Name: using eggs from schistosoma mansoni as an in vivo model of helminth-induced lung inflammation
Uploaded: 2012-06-05T13:00:00
Description: Watch this Scientific Journal Video about Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation at JoVE.com

Dette arbejde blev støttet af National Institutes of Health (NIH) AI091759 og Crohns og Colitis Foundation of America &#39;s William og Shelby Modell Family Foundation Research Award (til MN). Schistosome-inficerede mus blev leveret af NIAID Schistosome Resource Center (NIAID Kontrakt N01 A130026).

1. Oprensning Schistosoma mansoni-æg <ol><li> Infect schweizisk-Webster-mus med schistosome cercariae, som er infektiøse stadium i Schistosoma livscyklus 8. Alternativt, få schistosome-inficerede mus fra NIAID Schistosomiasis Resource Center ( <a href="http://www.schisto-resource.org/" target="_blank">http://www.schisto-resource.org/~~V</a> ). Efterlade mus 6-7 uger efter infektion, på hvilket tidspunkt-punkt æg er til stede i leveren og kan udvindes som beskrevet nedenfor (se <strong...

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	<li>World Health Organization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+and+control+of+schistosomiasis+and+soil-transmitted+helminthiasis:+report+of+a+WHO+expert+committee.">Prevention and control of schistosomiasis and soil-transmitted helminthiasis: report of a WHO expert committee.</a> WHO Technical Report Series 912. , (2002).</li><li>Pearce, E. J., MacDonald, A. S. <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+schistosomiasis.">The immunobiology of schistosomiasis.</a> Nat. Rev. Immunol. 2, 499-511 (2002).</li><li>Sandler, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+gene+expression+profiles+during+acute+pathogen-induced+pulmonary+inflammation+reveal+divergent+roles+for+Th1+and+th2+responses+in+tissue+repair.">Global gene expression profiles during acute pathogen-induced pulmonary inflammation reveal divergent roles for Th1 and th2 responses in tissue repair.</a> J. Immunol. 171, 3655-3667 (2003).</li><li>Perrigoue, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-31-IL-31R+interactions+negatively+regulate+type+2+inflammation+in+the+lung.">IL-31-IL-31R interactions negatively regulate type 2 inflammation in the lung.</a> J. Exp. Med. 204, 481-487 (2007).</li><li>Sandor, M., Weinstock, J. V., Wynn, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Granulomas+in+schistosome+and+mycobacterial+infections:+a+model+of+local+immune+responses.">Granulomas in schistosome and mycobacterial infections: a model of local immune responses.</a> Trends Immunol. 24, 44-52 (2003).</li><li>Nair, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alternatively+activated+macrophage-derived+RELM-{alpha}+is+a+negative+regulator+of+type+2+inflammation+in+the+lung.">Alternatively activated macrophage-derived RELM-{alpha} is a negative regulator of type 2 inflammation in the lung.</a> J. Exp. Med. 206, 937-952 (2009).</li><li>Dalton, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+the+isolation+of+schistosome+eggs+and+miracidia+free+of+contaminating+host+tissues.">A method for the isolation of schistosome eggs and miracidia free of contaminating host tissues.</a> Parasitology. 115, 29-32 (1997).</li><li>Lewis, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+laboratory+maintenance+of+Schistosoma+mansoni,+with+observations+on+three+schistosome/snail+host+combinations.">Large-scale laboratory maintenance of Schistosoma mansoni, with observations on three schistosome/snail host combinations.</a> J. Parasitol. 72, 813-829 (1986).</li><li>Tilney, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+lymphatic+drainage+in+the+adult+laboratory+rat.">Patterns of lymphatic drainage in the adult laboratory rat.</a> J. Anat. 109 (Pt. 3), 369-383 (1971).</li><li>Lewis, F., Coligan, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Schistosomiasis.">Schistosomiasis.</a> Current protocols in immunology. Chapter 19, Unit 19 (2001).</li><li>Kaplan, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Th2+cells+are+required+for+the+Schistosoma+mansoni+egg-induced+granulomatous+response.">Th2 cells are required for the Schistosoma mansoni egg-induced granulomatous response.</a> J. Immunol. 160, 1850-1856 (1998).</li><li>Wynn, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibrotic+disease+and+the+T(H)1/T(H)2+paradigm.">Fibrotic disease and the T(H)1/T(H)2 paradigm.</a> Nat. Rev. Immunol. 4, 583-594 (2004).</li><li>Cheever, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variation+of+hepatic+fibrosis+and+granuloma+size+among+mouse+strains+infected+with+Schistosoma+mansoni.">Variation of hepatic fibrosis and granuloma size among mouse strains infected with Schistosoma mansoni.</a> Am. J. Trop. Med. Hyg. 37, 85-97 (1987).</li></ol>

Her er en model anvender Schistosoma mansoni-æg at inducere type 2 lungeinflammation beskrevet. Karakteristiske træk ved denne model omfatter potente Th2-immunrespons, luftvejsinflammation og pulmonær granuloma-dannelse. Eftersom disse parametre afhænger CD4 Th2-celle-responser 11, er en nyttig model til at undersøge virkningerne af protein-specifikke eller slægt-specifikke deletioner i Th2-celle-responser. Her udlæsninger omfatter kvantificering af antigen-specifikke Th2-cytokiner, analyse af...

<table border="1"><tbody><tr><td> Reagens </td><td> Størrelse / Beskrivelse </td><td> Firma </td><td> Katalog # </td></tr><tr><td> Collagenase / dispase </td><td> 500 mg </td><td> Roche Applied Science </td><td> 11 097 113 001 </td></tr><tr><td> Diff Quik Hema 3 Stain </td><td> Pak </td><td> Fisher Scientific Co LLC </td><td> 122-911 </td></tr><tr><td> DMEM </td><td> Flydende </td><td> Gibco </td><td> 11965 </td></tr><tr><td> Falcon 50mL </td><td> Pak </td><td> Falcon </td><td> 352070 </td></tr><tr><td> Intramedic PE slange </td><td> 0,58 mm </td><td> Becton Dickinson og Co </td><td> 427410 </td></tr><tr><td> Isofluran </td><td> 250 ml </td><td> Abbot Dyresundhed </td><td> 52600405 </td></tr><tr><td> Ketamin </td><td> 100 mg / ml </td><td> Fort Dodge Animal Health </td&gt; <td> N / A </td></tr><tr><td> Mannose-receptor-antistof </td><td> Biotin </td><td> ABD Serotec </td><td> MCA2235B </td></tr><tr><td> Metallic Strainer </td><td> Standard køkken si </td><td> Mål </td><td> Laurentiis </td></tr><tr><td> Paraformaldehyd </td><td> 16% </td><td> Electron Microsc. Sciences </td><td> 15710 </td></tr><tr><td> Penicillin / streptomycin </td><td> 100x </td><td> Gibco </td><td> 15070 </td></tr><tr><td> Percoll </td><td> 1 Liter </td><td> GE Healthcare Biosciences </td><td> 17 0891 01 </td></tr><tr><td> RELMα antistof </td><td> 50 ug </td><td> Peprotech </td><td> 500 P214 </td></tr><tr><td> Kirurgisk Tråd </td><td> 2,0 USP 100 yds </td><td> Kirurgiske specialer Corp </td><td> SP118 </td></tr><tr><td> Kanyle </td><td> 23G, 3/4 inch </td><td> BD Vacutainer Lab. Med. </td><td>305143 </td></tr><tr><td> USA standardtestprogrammet Sieve </td><td> 150 um, 0,0059 &quot; </td><td> WS Tyler, Inc. </td><td> ASTME-11, Spec. Nr. 100 </td></tr><tr><td> Xylazin </td><td> 20 mg / ml </td><td> Butler </td><td> N / A </td></tr></tbody></table>

using eggs from schistosoma mansoni as an in vivo model of helminth-induced lung inflammation

Schistosoma parasitter er blod ikter, der inficerer skønsmæssigt 200 millioner mennesker verden over 1.. Ved kronisk infektion med Schistosoma, den alvorlige patologi, herunder leverfibrose og splenomegali, er forårsaget af immunrespons parasitæg stedet parasitten selve 2. Parasitæg inducere et Th2-respons er kendetegnet ved produktion af IL-4, IL-5 og IL-13, den alternative aktivering af makrofager og rekrutteringen af ​​eosinofiler. Her beskriver vi injektion af Schistosoma mansoni-æg som en model til at undersøge parasit-specifikke Th2 cytokin responser i lungen og drænende lymfeknuder, dannelsen af pulmonale granulomer omgiver ægget og luftvejsbetændelse. Efter intraperitoneal sensibilisering og intravenøs udfordring, S. mansoni-æg transporteres til lungen via den pulmonale arterier, hvor de bliver fanget i lungeparenkymved granulomer sammensat af lymfocytter, eosinofiler og alternativt aktiverede makrofager 3-6. Forbundet med granuloma-dannelse, inflammation i broncho-alveolare rum, ekspansion af de drænende lymfeknuder og CD4 T-celleaktivering kan observeres. Her har vi detaljeret protokol for at isolere Schistosoma mansoni-æg fra inficerede levere (ændret fra 7), sensibiliserende og udfordrende mus, og udvinding af de organer (broncho-alveolær lavage (BAL), lunge-og drænende lymfeknuder) til analyse. Vi har også repræsentant histologisk og immunologiske data og forslag til yderligere immunologiske analyse. Samlet Denne fremgangsmåde tilvejebringer en in vivo model til undersøgelse af helminth-inducerede immunologiske responser i lungen, som er bredt anvendelig til undersøgelse af Th2 inflammatoriske sygdomme, herunder helmint infektion, fibrotiske sygdomme, allergisk inflammation og astma. Fordele ved denne model til studiet af type to inflammation i lungen omfatter reproducerbarhed en potent Th2 inflammatorisk respons i lungerne og drænende lymfeknuder, den lette evaluering af inflammation ved histologisk undersøgelse af granulomer omkring ægget, og potentialet for langtidslagring af parasitten æg.

Watch this Scientific Journal Video about Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation at JoVE.com

Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation

Schistosoma mansoni-æg er potente stimulatorer af T-hjælpercelle type 2 (Th2) immunrespons, der er karakteristisk for parasit-infektion, astma og allergisk inflammation. Denne protokol anvender S. mansoni æg injektion til frembringelse af en CD4 Th2 cytokin-induceret inflammatorisk respons i lungerne, kendetegnet ved lunge granulomdannelse omkring ægget, eosinofili og makrofag alternativ aktivering.

Brug Æg fra<em&gt; Schistosoma mansoni</em&gt; Som en<em&gt; In vivo</em&gt; Model for helminth-induceret lungeinflammation

Schistosoma parasites are blood flukes that infect an estimated 200 million people worldwide 1. In chronic infection with Schistosoma, the severe ...

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Immunology and Infection

Using Eggs from Schistosoma mansoni as an In vivo Model of Helminth-induced Lung Inflammation

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