Name: trichuris muris infection: a model of type 2 immunity and inflammation in the gut
Uploaded: 2011-05-24T12:00:00
Description: Watch this Scientific Journal Video about Trichuris muris Infection: A Model of Type 2 Immunity and Inflammation in the Gut at JoVE.com

Detta arbete stöddes av den kanadensiska Institutes of Health Research (MSH-95.368, MOP-89.773 och MOP-106.623 till CZ) och Kanada Stiftelsen för innovation. SCM är mottagare av en CIHR / kanadensiska Association of Gastroenterology postdoktorsstipendium. CZ är ett CIHR ny utredare.

1. Uppförökningsmaterial Trichuris Muris ägg <ol><li> Att generera nya partier av Trichuris Muris ägg, infektera 20-30 immundefekta möss (t.ex. NOD.Cg-Prkdc Scid Il2rg tm1Wjl / SzJ (NSG) eller 129S6/SvEvTac-Rag2 tm1Fwa (RAG2 - / -)) eller genetiskt känsliga möss ( t ex. AKR / J) som är 6-8 veckor gamla med ca 300 Trichuris Muris ägg via oral sondmatning. </li><li> Efter 32-35 dagar offra de möss med CO 2 kvävning. </li><li> Ex...

<ol>
	<li>Cliffe, L. J., Grencis, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Trichuris+muris+system:+a+paradigm+of+resistance+and+susceptibility+to+intestinal+nematode+infection.">The Trichuris muris system: a paradigm of resistance and susceptibility to intestinal nematode infection.</a> Adv. Parasitol. 57, 255-307 (2004).</li><li>Else, K. J., Finkelman, F. D., Maliszewski, C. R., Grencis, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytokine-mediated+regulation+of+chronic+intestinal+helminth+infection.">Cytokine-mediated regulation of chronic intestinal helminth infection.</a> J. Exp. Med. 179, 347-351 (1994).</li><li>Bancroft, A. J., Grencis, R. 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Med. 203, 843-849 (2006).</li><li>Artis, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RELM&#946;/FIZZ2+is+a+goblet+cell-specific+immune-effector+molecule+in+the+gastrointestinal+tract.">RELM&#946;/FIZZ2 is a goblet cell-specific immune-effector molecule in the gastrointestinal tract.</a> Proc. Natl. Acad. Sci. U. S. A. 101, 13596-13600 (2004).</li><li>Datta, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+novel+genes+in+intestinal+tissue+that+are+regulated+after+infection+with+an+intestinal+nematode+parasite.">Identification of novel genes in intestinal tissue that are regulated after infection with an intestinal nematode parasite.</a> Infect. Immun. 73, 4025-4033 (2005).</li><li>Cliffe, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+intestinal+epithelial+cell+turnover:+a+new+mechanism+of+parasite+expulsion.">Accelerated intestinal epithelial cell turnover: a new mechanism of parasite expulsion.</a> Science. 308, 1463-1465 (2005).</li><li>Artis, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+weapons+in+the+war+on+worms:+identification+of+putative+mechanisms+of+immune-mediated+expulsion+of+gastrointestinal+nematodes.">New weapons in the war on worms: identification of putative mechanisms of immune-mediated expulsion of gastrointestinal nematodes.</a> Int. J. Parasitol. 36, 723-733 (2006).</li><li>Finkelman, F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytokine+regulation+of+host+defense+against+parasitic+gastrointestinal+nematodes:+lessons+from+studies+with+rodent+models.">Cytokine regulation of host defense against parasitic gastrointestinal nematodes: lessons from studies with rodent models.</a> Annu. Rev. Immunol. 15, 505-533 (1997).</li><li>Grencis, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enteric+helminth+infection:+immunopathology+and+resistance+during+intestinal+nematode+infection.">Enteric helminth infection: immunopathology and resistance during intestinal nematode infection.</a> Chem. Immunol. 66, 41-61 (1997).</li><li>Khan, W. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+intestinal+muscle+contraction+by+interleukin-9+(IL-9)+or+IL-9+neutralization:+correlation+with+worm+expulsion+in+murine+nematode+infections.">Modulation of intestinal muscle contraction by interleukin-9 (IL-9) or IL-9 neutralization: correlation with worm expulsion in murine nematode infections.</a> Infect. Immun. 71, 2430-2438 (2003).</li><li>Khan, W. I., Blennerhasset, P., Ma, C., Matthaei, K. I., Collins, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stat6+dependent+goblet+cell+hyperplasia+during+intestinal+nematode+infection.">Stat6 dependent goblet cell hyperplasia during intestinal nematode infection.</a> Parasite Immunol. 23, 39-42 (2001).</li><li>Akiho, H., Blennerhassett, P., Deng, Y., Collins, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+IL-4,+IL-13,+and+STAT6+in+inflammation-induced+hypercontractility+of+murine+smooth+muscle+cells.">Role of IL-4, IL-13, and STAT6 in inflammation-induced hypercontractility of murine smooth muscle cells.</a> Am. J. Physiol. Gastrointest. Liver Physiol. 282, 226-2232 (2002).</li><li>Kopper, J. J., Mansfield, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+improved+methods+for+delivery+of+Trichuris+muris+to+the+laboratory+mouse.">Development of improved methods for delivery of Trichuris muris to the laboratory mouse.</a> Parasitol. Res. 107, 1103-1113 (2010).</li><li>Bancroft, A. J., Else, K. J., Grencis, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-level+infection+with+Trichuris+muris+significantly+affects+the+polarization+of+the+CD4+response.">Low-level infection with Trichuris muris significantly affects the polarization of the CD4 response.</a> Eur. J. Immunol. 24, 3113-3118 (1994).</li></ol>

Detta protokoll detaljer en standard hög dos akut Trichuris Muris infektion som kan ändras efter behov av prövaren. Till exempel kan mössen offras och vävnader skördas på olika dagar. Att fastställa att de möss med framgång har etablerat fullt mask börda de kan offras på dag 14, då alla möss bör göra en börda på cirka 200 maskar. Möss kan också smittas i 32 dagar där någon mask upptäcks har nått mognad och skulle stanna hos värd under hela dess liv. Dessutom kan protokollet anpassas til...

<table border="1"><tbody><tr><th> Reagensets namn </th><th> Företag </th><th> Katalognummer </th><th> Kommentarer </th></tr><tr><td> Utfodring av djur Nålar (18 x 1 ½ &quot;) </td><td> Popper </td><td> 7912 </td><td></td></tr><tr><td> Jämnt svängt Tång </td><td> Roboz </td><td> RS-5047 </td><td></td></tr><tr><td> DMEM </td><td> Gibco </td><td> 11.965 </td><td></td></tr><tr><td> NOD.Cg-Prkdc Scid Il2r gtm1Wjl / SzJ (NSG) </td><td> Jackson Laboratories </td><td> 005.557 </td><td> Dessa är de möss vi använde dock bör alla immunbrist möss eller känsliga stam arbete. </td></tr><tr><td> RNAlater </td><td> Qiagen </td><td> 76.104 </td><td></td></tr><tr><td> 2 ml rör </td><td> Axygen </td><td> MCT-200-C </td><td></td></tr><tr><td> 15 ml rör </td><td> Falcon </td><td> 352.096 </td><td></td></tr><tr><td> 6 brunnar </td><td> Falcon </td><td> 353.046 </td><td></td></tr><tr><td> Paraformaldehyd </td><td> Elektronmikroskopi Vetenskap </td><td> 15.710 </td><td></td></tr><tr><td> α-RELMβ antikropp </td><td> PeproTech Inc </td><td> 0694270Rb </td><td></td></tr></tbody></table>

trichuris muris infection: a model of type 2 immunity and inflammation in the gut

Trichuris Muris är en naturlig patogen av möss och är biologiskt och antigena liknande arter av Trichuris som infekterar människor och djur 1. Infektiösa ägg ges via oral sondmatning, lucka i distala tunntarmen, invadera tarmcellerna (IECS) som kantar kryptor i blindtarmen och proximala kolon och på mognad maskarna frigöra ägg i miljön 1. Denna modell är ett kraftfullt verktyg för att undersöka faktorer som styr CD4 + T-hjälparceller (Th) celler aktivering samt förändringar i intestinal epitel. Det immunsvar som sker i resistenta inavlade stammar, såsom C57BL / 6 och BALB / c, kännetecknas av Th2 polariserad cytokiner (IL-4, IL-5 och IL-13) och utvisning av maskar medan Th1-associerade cytokiner (IL -12, IL-18, IFN-γ) främja kroniska infektioner hos genetiskt känsliga AKR / möss J 2-6. Th2 cytokiner främja fysiologiska förändringar i intestinal mikromiljön bland annat snabb omsättning av IECS, bägare celldifferentiering, rekrytering och förändringar i epitelial permeabilitet och smidig muskel kontraktion, som alla har varit inblandade i masken utvisning 7-15. Här har vi närmare ett protokoll för att sprida Trichuris Muris ägg som kan användas i kommande experiment. Vi tillhandahåller också en skörd prov experimentell med förslag för post-infektion analys. Sammantaget kommer detta protokoll att ge forskare med grundläggande verktyg för att utföra en Trichuris Muris modell musen infektion som kan användas för att behandla frågor som rör Th benägenhet i mag-tarmkanalen samt immunsystemets effektorsteg av IECS.

Watch this Scientific Journal Video about Trichuris muris Infection: A Model of Type 2 Immunity and Inflammation in the Gut at JoVE.com

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

Trichuris Muris infektion är en intestinal modell av Th2 immuniteten, om resistenta möss bildar ett skyddande Th2 respons och mottagliga möss generera en patologisk Th1 svar.

Trichuris Muris Infektion: En modell av typ 2 Immunitet och inflammation i tarmen

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

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Immunology and Infection

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

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