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>
	<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. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Th1+and+Th2+cells+and+immunity+to+intestinal+helminths.">Th1 and Th2 cells and immunity to intestinal helminths.</a> Chem. Immunol. 71, 192-208 (1998).</li><li>Bancroft, A. J., McKenzie, A. N., 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=A+critical+role+for+IL-13+in+resistance+to+intestinal+nematode+infection.">A critical role for IL-13 in resistance to intestinal nematode infection.</a> J. Immunol. 160, 3453-3461 (1998).</li><li>Helmby, H., Takeda, K., Akira, S., 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=Interleukin+(IL)-18+promotes+the+development+of+chronic+gastrointestinal+helminth+infection+by+downregulating+IL-13.">Interleukin (IL)-18 promotes the development of chronic gastrointestinal helminth infection by downregulating IL-13.</a> J. Exp. Med. 194, 355-364 (2001).</li><li>Owyang, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interleukin+25+regulates+type+2+cytokine-dependent+immunity+and+limits+chronic+inflammation+in+the+gastrointestinal+tract.">Interleukin 25 regulates type 2 cytokine-dependent immunity and limits chronic inflammation in the gastrointestinal tract.</a> J. Exp. 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>

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. ...

trichuris-muris-infection-model-type-2-immunity-inflammation

Research

JoVE Journal

Immunology and Infection

Capsular Serotyping of Streptococcus pneumoniae by Latex Agglutination

Latex agglutination reagents are widely used in microbial diagnosis, identification and serotyping. Streptococcus pneumoniae (the pneumococcus) is a major cause of morbidity and mortality world-wide. Current vaccines target the pneumococcal capsule, and there are over 90 capsular serotypes. Serotyping pneumococcal isolates is therefore important for assessing the impact of vaccination programs and for epidemiological purposes. The World Health Organization has recommended latex agglutination as an alternative method to the &#8216;gold standard&#8217; Quellung test for serotyping pneumococci. Latex agglutination is a relatively simple, quick and inexpensive method; and is therefore suitable for resource-poor settings as well as laboratories with high-volume workloads. Latex agglutination reagents can be prepared in-house utilizing commercially-sourced antibodies that are passively attached to latex particles. This manuscript describes a method of production and quality control of latex agglutination reagents, and details a sequential testing approach which is time- and cost-effective. 
This method of production and quality control may also be suitable for other testing purposes.

Capsular Serotyping of Streptococcus pneumoniae by Latex Agglutination

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We describe a method for generating Precision-cut Lung Slices (PCLS) and immunostaining them to visualize the localization of various immune cell types in the lung. Our protocol can be extended to visualize the location and function of many different cell types under a variety of conditions.

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Effects of Taste Signaling Protein Abolishment on Gut Inflammation in an Inflammatory Bowel Disease Mouse Model

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Here we present a protocol to investigate the effect of the nullification of gustation-related genes on immune responses in a dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) mouse model.

Inflammatory bowel disease (IBD) is one of the immune-related gastrointestinal disorders, including ulcerative colitis and Crohn's disease, that affects ...

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Virus spreading is a major cause of epidemic diseases. Thus, understanding the interaction between the virus and the host is very important to extend our knowledge of prevention and treatment of viral infection. The fruit fly Drosophila melanogaster has proven to be one of the most efficient and productive model organisms to screen for antiviral factors and investigate virus-host interaction, due to powerful genetic tools and highly conserved innate immune signaling pathways. The procedure described here demonstrates a nano-injection method to establish viral infection and induce systemic antiviral responses in adult flies. The precise control of the viral injection dose in this method enables high experimental reproducibility. Protocols described in this study include the preparation of flies and the virus, the injection method, survival rate analysis, the virus load measurement, and an antiviral pathway assessment. The influence effects of viral infection by the flies' background were mentioned here. This infection method is easy to perform and quantitatively repeatable; it can be applied to screen for host/viral factors involved in virus-host interaction and to dissect the crosstalk between innate immune signaling and other biological pathways in response to viral infection.

Establishment of Viral Infection and Analysis of Host-Virus Interaction in Drosophila Melanogaster

This protocol describes how to establish viral infection in vivo in Drosophila melanogaster using the nano-injection method and basic techniques to analyze virus-host interaction.

Virus spreading is a major cause of epidemic diseases. Thus, understanding the interaction between the virus and the host is very important to extend our ...

The Galleria mellonella Waxworm Infection Model for Disseminated Candidiasis

Candida species are common fungal commensals of humans colonizing the skin, mucosal surfaces, and gastrointestinal tract. Under certain conditions, Candida can overgrow their natural niches resulting in debilitating mucosal infections as well as life-threatening systemic infections, which are a major focus of investigation due to their associated high mortality rates. Animal models of disseminated infection exist for studying disease progression and dissecting the characteristics of Candida pathogenicity. Of these, the Galleria mellonella waxworm infection model provides a cost-effective experimental tool for high-throughput investigations of systemic virulence. Many other bacterial and eukaryotic infectious agents have been effectively studied in G. mellonella to understand pathogenicity, making it a widely accepted model system. Yet, variation in the method used to infect G. mellonella can alter phenotypic outcomes and complicate interpretation of the results. Here, we outline the benefits and drawbacks of the waxworm model to study systemic Candida pathogenesis and detail an approach to improve reproducibility. Our results highlight the range of mortality kinetics in G. mellonella and describe the variables which can modulate these kinetics. Ultimately, this method stands as an ethical, rapid, and cost-effective approach to study virulence in a model of disseminated candidiasis.

The Galleria mellonella Waxworm Infection Model for Disseminated Candidiasis

Galleria mellonella serves as an invertebrate model for disseminated candidiasis. Here, we detail the infection protocol and provide supporting data for the model&#39;s effectiveness.

Candida species are common fungal commensals of humans colonizing the skin, mucosal surfaces, and gastrointestinal tract. Under certain conditions, ...