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

This work was supported by the Canadian Institutes of Health Research (MSH-95368, MOP-89773 and MOP-106623 to C.Z.) and the Canada Foundation for Innovation. S.C.M. is the recipient of a CIHR/Canadian Association of Gastroenterology postdoctoral fellowship. C.Z. is a CIHR New Investigator.

1. Propagating Trichuris muris eggs
<ol>
<li>To generate new batches of Trichuris muris eggs, infect 20-30 immunodeficient mice (eg. NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ 
(NSG) or 129S6/SvEvTac-Rag2tm1Fwa (RAG2-/-)) or genetically susceptible mice (eg. AKR/J) that are 6-8 weeks old with approximately 300 Trichuris 
muris eggs by oral gavage.</li> 
<li>After 32&#x2013;35 days sacrifice the mice by CO2 asphyxiation.</li>
<li>Expose the ventral ...

<ol>
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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. 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This protocol details a standard high dose acute Trichuris muris infection which can be modified as required by the investigator. For example, mice can be sacrificed and tissues harvested on different days. To determine that the mice have successfully established full worm burden they can be sacrificed on day 14, at which point all mice should carry a burden of approximately 200 worms. Mice can also be infected for 32 days where any worm detected will have reached maturity and would remain with host for the d...

<table border="1">
	<tbody>
 <tr>
 	<th>Name of the reagent</th>
 <th>Company</th>
 <th>Catalogue number</th>
 <th>Comments</th>
 </tr>
 <tr>
 	<td>Animal Feeding Needles (18 x 1&frac12;")</td>
 <td>Popper</td>
 <td>7912</td>
 <td>&nbsp;</td>
 </tr>
		<tr>
 	<td>Smooth curved Forceps</td>
 <td>Roboz</td>
 <td>RS-5047</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 	<td>DMEM</td>
 <td>Gibco</td>
 <td>11965</td>
 <td>&nbsp;</td>
 </tr>
		<tr>
 	<td>NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG)</td>
 <td>Jackson Laboratories</td>
 <td>005557</td>
 <td>These are the mice we used, however, any immunodeficient mice or susceptible strain should work.</td>
 </tr>
 <tr>
 	<td>RNAlater</td>
 <td>Qiagen</td>
 <td>76104</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 	<td>2 ml tubes</td>
 <td>Axygen</td>
 <td>MCT-200-C</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 	<td>15 ml tubes</td>
 <td>Falcon</td>
 <td>352096</td>
 <td>&nbsp;</td>
 </tr><tr>
 	<td>6 well plates</td>
 <td>Falcon</td>
 <td>353046</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 	<td>Paraformaldehyde</td>
 <td>Electron Microscopy Science</td>
 <td>15710</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 	<td>&alpha;-RELM&beta; antibody</td>
 <td>PeproTech Inc</td>
 <td>0694270Rb</td>
 <td>&nbsp;</td>
 </tr>
 </tbody>
</table>

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.

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 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 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. ...

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