Rapid dendritic cell mobilization to the large intestinal epithelium is associated with resistance to Trichuris muris infection - PubMed (original) (raw)
Comparative Study
. 2009 Mar 1;182(5):3055-62.
doi: 10.4049/jimmunol.0802749.
Matthew L Deschoolmeester, Marcus Svensson, Gareth Howell, Aikaterini Bazakou, Larisa Logunova, Matthew C Little, Nicholas English, Matthias Mack, Richard K Grencis, Kathryn J Else, Simon R Carding
Affiliations
- PMID: 19234202
- PMCID: PMC2671799
- DOI: 10.4049/jimmunol.0802749
Comparative Study
Rapid dendritic cell mobilization to the large intestinal epithelium is associated with resistance to Trichuris muris infection
Sheena M Cruickshank et al. J Immunol. 2009.
Abstract
The large intestine is a major site of infection and disease, yet little is known about how immunity is initiated within this site and the role of dendritic cells (DCs) in this process. We used the well-established model of Trichuris muris infection to investigate the innate response of colonic DCs in mice that are inherently resistant or susceptible to infection. One day postinfection, there was a significant increase in the number of immature colonic DCs in resistant but not susceptible mice. This increase was sustained at day 7 postinfection in resistant mice when the majority of the DCs were mature. There was no increase in DC numbers in susceptible mice until day 13 postinfection. In resistant mice, most colonic DCs were located in or adjacent to the epithelium postinfection. There were also marked differences in the expression of colonic epithelial chemokines in resistant mice and susceptible mice. Resistant mice had significantly increased levels of epithelium-derived CCL2, CCL3, CCL5, and CCL20 compared with susceptible mice. Furthermore, administering neutralizing CCL5 and CCL20 Abs to resistant mice prevented DC recruitment. This study provides clear evidence of differences in the kinetics of DC responses in hosts inherently resistant and susceptible to infection. DC responses in the colon correlate with resistance to infection. Differences in the production of DC chemotactic chemokines by colonic epithelial cells in response to infection in resistant vs susceptible mice may explain the different kinetics of the DC response.
Figures
Figure 1. Rapid Recruitment of Colonic DCs in T. muris infected Resistant Mice
Colonic lamina propria cells were identified by flow cytometry using CD45, CD11c and MHC II. Data shows representative profile of CD11c and CD45 expression (A). The region drawn around CD45+CD11c+ cells was used to determine MHCII expression (B). (C) shows the number of DCs in BALB/c and AKR mice pre (0) and post-infection with 150 (HD BALB/c; AKR) or 20 embryonated T.muris eggs (LD BALB/c). (D) shows the subtypes of colonic DCs in AKR and BALB/c mice. The data is from 4-9 experiments. *p<0.01, **p<0.001 comparing resistant versus susceptible mice at the same time point.
Figure 2. Altered distribution of Colonic DCs in Resistant but not Susceptible Mice
Colon from resistant BALB/c mice (HD BALB/c: A-D), susceptible BALB/c mice (LD BALB/c; E, F) susceptible AKR (G-K) at D0 (A and G) and D1 (B, E and H), D7 (C, F and I) and D13 post-infection (J) were stained for DCs (CD11c, red) and epithelial cells (cytokeratin, green) and counterstained with TOPRO3 or DAPI (blue). Isotype controls for BALB/c (D) and AKR (K) tissue are shown. Magnification x 400. (L) The numbers of DCs/crypt associated with the epithelium of AKR and BALB/c mice pre- and post-infection was quantified.
Figure 3. DC -Epithelial Cell Interactions in Infected Resistant Mice
Immunofluorescent confocal images (A, B, C, E and F) and electron micrograph images (D) of colon from resistant BALB/c mice at D1 (A, B, C, E and F) and D7 (D) post-infection (BALB/c HD) showing DCs within the colonic epithelium. (A-C) Tissue was stained with claudin 3 (red) and CD11c (green) and counterstained with TOPRO3 (Blue). (A) identifies a large DC aggregate underneath the colonic epithelium with two DCs in the epithelial layer (boxed area), (B) is a 3D render of a Z-stacked image of the boxed area in (A) showing the X, Y and Z dimensions and identifies a dendrite extending through the epithelium and into the lumen (arrowed). The points of co-localisation between the DC and claudin 3 are highlighted in white in (C) and are arrowed. (D) Electron microscope photomicrographs of T. muris infected BALB/c colon showing a DC localized to epithelial tight junctions between adjacent epithelial cells (EC) and extending dendrites (arrowed) into the lumen. In (D) a DC appears to be taking up cellular antigen (red arrow), possibly apoptotic cell material (Mag. x 6900). (E and F) Confocal images of colon from resistant BALB/c mice stained with cytokeratin (green, epithelial cells), CD11c (red, DCs) and counterstained with TOPRO3 (blue). (E) is the original unmanipulated confocal image in (E) the inset box shows the DC in isolation with the green and blue fluorescence subtracted out to reveal DC morphology. (F) is an orthogonal view of a full thickness (Z-stack) image of the (right hand panel) showing the DC touching adjacent epithelial cells (Grid size 5 μm2).
Figure 4. In Response to Infection DCs Cross the Epithelial Basement Membrane
Immunofluorescent images of sections of colon from resistant BALB/c mice before (A, B) and 1 day post-infection (C-E) with a high antigen dose of T. muris (BALB/c HD). Sections of colon were stained for DC (CD11c, green), basement membrane (laminin, red) and counterstained with DAPI (blue) (A-D). In (A and C) are the original images over-exposed to visualise autofluorescent epithelial cells (red). (B and D) are higher magnification of the areas boxed in A,C showing DCs beneath the epithelial basement membrane prior to infection (B) but after infection they are breaching the basement membrane and are in the epithelial layer (D, E). _L_= lumen, e-epithelium. (E) shows iso-surface volume rendering of the confocal z-sections from (C). Grid squares = 5μm2. Iso-surface volume rendering was carried out with the Surpass module within Imaris v5.7 (Bitplane AG, Switzerland).
Figure 5. Distinctive Colonic DCs in infected Resistant and Susceptible Mice
MHCII (A), CD86 (B), TLR2 (C), TLR4 (D), CCR7 (E) expression and FITC Dextran uptake (F) was analysed by flow cytometry by colonic DCs from BALB/c (filled bars) and AKR (open bars) mice pre (0) and post-infection with 150 or 20 embryonated T. muris eggs (hatched bars). (G) The proportion of CD11c+MHCII+ DCs were analysed in the MLNs of BALB/c and AKR mice during the time course of infection. The data is from 4-9 experiments. *p<0.05; **p<0.01;***p<0.001 P values comparing resistant versus susceptible mice at the same timepoint and in E and F comparing naïve mice and D7 post-infection.
Figure 6. Increased Production of DC chemoattractants by Colonic Epithelial Cells in Resistant Mice Promote DC recruitment
Epithelial cells (n=4) from the proximal colons of resistant BALB/c, susceptible AKR and susceptible BALB/c mice were analysed for expression of CCL2, CCL3, CCL5, CCL20 and TSLP mRNA 0, and 24 hours (A) and 7 days post-infection (B) by q-PCR. The data is shown as the relative expression of mRNA compared to samples from naïve non-infected animals. (C) In order to assess whether CCL5 and CCL20 were necessary for DC recruitment in resistant BALB/c mice, BALB/c mice were infected with T.muris and given intravenous neutralising antibodies to CCL5 and CCL20 (left hand column) or control rat IgG (right hand column). After 24 hours the DCs were enumerated by flow cytometry. In antibody treated mice there was a complete ablation of DC recruitment (***p=0.0001, n=8).
References
- Rakoff-Nahoum S, Paglino J, Eslami-Varzaneh F, Edberg S, Medzhitov R. Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis. Cell. 2004;118:229–241. - PubMed
- Singh JC, Cruickshank SM, Newton DJ, Wakenshaw L, Graham A, Lan J, Lodge JP, Felsburg PJ, Carding SR. Toll-like receptor-mediated responses of primary intestinal epithelial cells during the development of colitis. American Journal of Physiology - Gastrointestinal & Liver Physiology. 2005;288:G514–524. - PubMed
Publication types
MeSH terms
LinkOut - more resources
Full Text Sources
Other Literature Sources