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 arbeidet ble støttet av National Institutes of Health (NIH) AI091759 og Crohns og kolitt Foundation of America William og Shelby Modell Family Foundation Research Award (til MN). Schistosome-infiserte mus ble levert av NIAID Schistosome Resource Center (NIAID Kontrakt N01 A130026).

1. Rensing Schistosoma mansoni Eggs <ol><li> Infisere sveitsisk-Webster mus med schistosome cercariae, som er smittsom scenen i Schistosoma livssyklus 8. Alternativt få schistosome-infiserte mus fra NIAID Schistosomiasis Resource Center ( <a href="http://www.schisto-resource.org/" target="_blank">http://www.schisto-resource.org/~~V</a> ). La mus for 6-7 uker etter smitte, og da-punkt, egg finnes i leveren og kan utvinnes som beskrevet under (se fig. 1). </li><li> Avlive m...

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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 modell ansette Schistosoma mansoni egg til å indusere type 2 lunge betennelse beskrevet. Karakteristiske trekk ved denne modellen inkluderer den potente Th2 immunforsvaret, luftveier betennelse og pulmonal svulstdannelser. Siden disse parametrene er avhengige av CD4 Th2 celle responser 11, er dette en nyttig modell for å undersøke effektene av protein-spesifikke eller avstamning-spesifikk slettinger på Th2 celle responser. Her lettleselig ville inkludere kvantifisering av antigen-spesif...

<table border="1"><tbody><tr><td> Reagens </td><td> Størrelse / beskrivelse </td><td> Firma </td><td> Catalog # </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> Pakk </td><td> Fisher Scientific Co LLC </td><td> 122-911 </td></tr><tr><td> DMEM </td><td> Flytende </td><td> Gibco </td><td> 11965 </td></tr><tr><td> Falcon 50ml </td><td> Pakk </td><td> Falcon </td><td> 352070 </td></tr><tr><td> Intramedic PE Tubing </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 Animal Health </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 reseptor antistoff </td><td> Biotin </td><td> Abd Serotec </td><td> MCA2235B </td></tr><tr><td> Metallic Sil </td><td> Standard kjøkken sil </td><td> Target </td><td> Laurentiis </td></tr><tr><td> Paraformaldehyde </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 biovitenskap </td><td> 17 0891 01 </td></tr><tr><td> RELMα antistoff </td><td> 50 mikrogram </td><td> Peprotech </td><td> 500-P214 </td></tr><tr><td> Kirurgisk Tråd </td><td> 2.0 USP 100 yds </td><td> Kirurgiske spesialiteter Corp </td><td> SP118 </td></tr><tr><td> Sprøytespissen </td><td> 23G, 3/4 tommers </td><td> BD Vacutainer Lab. Med. </td><td>305143 </td></tr><tr><td> USA Standard Testing Sieve </td><td> 150 mikrometer og 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 flukes som infiserer anslagsvis 200 millioner mennesker verden over 1. I kronisk infeksjon med Schistosoma, den alvorlige patologi, herunder leverfibrose og splenomegali, er forårsaket av immunrespons mot parasitten eggene heller enn parasitten selv to. Parasitt egg indusere en Th2 respons preget av produksjonen av IL-4, IL-5 og IL-13, alternativ aktivering av makrofager og rekruttering av eosinofile. Her beskriver vi injeksjon av Schistosoma mansoni egg som en modell for å undersøke parasitt-spesifikke Th2 cytokin respons i lungene og drenering lymfeknuter, dannelsen av pulmonale granulomer rundt egget og luftveier betennelser. Etter intraperitoneal sensibilisering og intravenøs utfordring, S. mansoni egg transporteres til lungene via lungene arteriene hvor de er fanget i lungene parenchymaav granulomer består av lymfocytter, eosinofile og alternativt aktiveres makrofager 3-6. Assosiert med svulstdannelser, betennelse i de broncho-alveolene mellomrom, utvidelse av drenering lymfeknuter og CD4 T-celleaktivering kan observeres. Her har vi detalj protokollen for å isolere Schistosoma mansoni egg fra infiserte leveren (modifisert fra 7), allergifremkallende og utfordrende mus, og utvinne organene (broncho-alveolar lavage (BAL), lunge og drenering lymfeknuter) for analyse. Vi inkluderer også representant histologisk og immunologiske data og forslag til ytterligere immunologisk analyse. Samlet gir denne metoden en in vivo modell for å undersøke helminth-indusert immunologiske reaksjoner i lungene, som er bredt gjelder for studiet av Th2-inflammatoriske sykdommer, inkludert helminth infeksjon, fibrotiske sykdommer, allergisk betennelse og astma. Fordelene med denne modellen for studiet av type to betennelse i lungene inkluderer reproduserbarhet av en potent Th2 betennelsesreaksjon i lungene og drenering lymfeknuter, den enkle vurdering av betennelse ved histologisk undersøkelse av granulomer rundt egget, og potensialet for langsiktig lagring av parasitten egg.

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 egg er potente stimulators av T hjelperen type 2 (Th2) immunrespons, karakteristisk for parasitt infeksjon, astma og allergisk betennelse. Denne protokollen benytter S. mansoni egg injeksjon for å generere et CD4 Th2 cytokin-indusert betennelsesreaksjon i lungene, preget av lunge svulstdannelser rundt egg, eosinofili og macrophage alternativ aktivering.

Bruke Egg fra<em&gt; Schistosoma mansoni</em&gt; Som en<em&gt; In vivo</em&gt; Modell av Helminth-indusert lungebetennelse

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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