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Proinflammatory microenvironments within the intestine regulate the differentiation of tissue-resident CD8⁺ T cells responding to infection.

Bergsbaken T, Bevan MJ - Nat. Immunol. (2015)

Bottom Line: CD103(-) T cells did not require transforming growth factor-β (TGF-β) signaling but were true resident memory cells.Unlike CD103(+)CD8(+) T cells, which were TGF-β dependent and were scattered in the tissue, CD103(-)CD8(+) T cells clustered with CD4(+) T cells and CX3CR1(+) macrophages and/or dendritic cells around areas of bacterial infection.CXCR3-dependent recruitment of cells to inflamed areas was critical for development of the CD103(-) population and pathogen clearance.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Howard Hughes Medical Institute, University of Washington, Seattle, Washington, USA.

ABSTRACT
We report that oral infection with Yersinia pseudotuberculosis results in the development of two distinct populations of pathogen-specific CD8(+) tissue-resident memory T cells (TRM cells) in the lamina propria. CD103(-) T cells did not require transforming growth factor-β (TGF-β) signaling but were true resident memory cells. Unlike CD103(+)CD8(+) T cells, which were TGF-β dependent and were scattered in the tissue, CD103(-)CD8(+) T cells clustered with CD4(+) T cells and CX3CR1(+) macrophages and/or dendritic cells around areas of bacterial infection. CXCR3-dependent recruitment of cells to inflamed areas was critical for development of the CD103(-) population and pathogen clearance. Our studies have identified the 'preferential' development of CD103(-) TRM cells in inflammatory microenvironments within the lamina propria and suggest that this subset has a critical role in controlling infection.

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Yptb-OVA infection stimulates formation of CD103– CD8 T cell clusters in the LP. C57BL/6 mice received 104 GFP OT-I T cells and were orally infected with Yptb-OVA the following day. The terminal ileum was isolated at 9 days post infection and tissue sections were analyzed by immunohistochemistry. (a) Distribution of OT-I T cells (green) in the ileum. Arrows indicate LP clusters near the crypts and open arrowheads indicate those in the upper part of the villi. Epithelial cells are stained with anti-EpCam antibody (red). Scale bar equals 100 μm. Representative image from 5 mice. (b) Tissue sections were stained with anti-CD103 antibody (red) to determine expression on OT-I T cells (green). Nuclei are stained with DAPI (blue). Open arrowheads indicate CD103+ OT-I T cells. Images of villous (left) and clustered (right) OT-I T cells. (c) OT-I clusters form around areas of Yptb-OVA infection (anti-Yptb-red, OT-I-green, nuclei-blue). (b-c) Representative images from 3 mice. Scale bars equal 25 μm.
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Figure 4: Yptb-OVA infection stimulates formation of CD103– CD8 T cell clusters in the LP. C57BL/6 mice received 104 GFP OT-I T cells and were orally infected with Yptb-OVA the following day. The terminal ileum was isolated at 9 days post infection and tissue sections were analyzed by immunohistochemistry. (a) Distribution of OT-I T cells (green) in the ileum. Arrows indicate LP clusters near the crypts and open arrowheads indicate those in the upper part of the villi. Epithelial cells are stained with anti-EpCam antibody (red). Scale bar equals 100 μm. Representative image from 5 mice. (b) Tissue sections were stained with anti-CD103 antibody (red) to determine expression on OT-I T cells (green). Nuclei are stained with DAPI (blue). Open arrowheads indicate CD103+ OT-I T cells. Images of villous (left) and clustered (right) OT-I T cells. (c) OT-I clusters form around areas of Yptb-OVA infection (anti-Yptb-red, OT-I-green, nuclei-blue). (b-c) Representative images from 3 mice. Scale bars equal 25 μm.

Mentions: Little is known about the distribution of antigen specific CD8 T cells upon colonization with an intestinal pathogen, and we hypothesized the phenotypic differences we observed may be due to different microenvironments created by Yptb infection within the intestine. We examined the localization of GFP OT-I T cells in the intestine of Yptb-OVA infected mice at 9 days post infection. The OT-I cells in the ileum of the small intestine could be divided into two groups based on their distribution: those that were present singly and evenly distributed throughout the tissue, and those that formed clusters (Fig. 4a). These CD8 T cell clusters were found primarily underlying the crypts; however, they could occasionally be found in the villus LP (Fig. 4a). We then examined the surface expression of CD103 on OT-I T cells within the intestine and found that while the majority of the uniformly distributed OT-I cells in IEL and LP expressed CD103 (Fig. 4b left), OT-I T cells in clusters near the crypts were mostly CD103– (Fig. 4b right, Supplementary Fig. 4a). Tissue sections were also stained with Yptb-specific antibodies, indicating OT-I T cells clustered around areas of bacterial infection (Fig. 4c). Similar CD8 T cell clusters were also found in the cecum and colon but not the duodenum (Supplementary Fig. 4b-d). This is consistent with CD8 T cell clusters indicating areas of LP infection, as Yptb infects the distal part of the small intestine, cecum, and colon, but not the proximal small intestine (Supplementary Fig. 4e). These data suggest recruitment to these intestinal aggregates during infection affects the phenotype and function of LP CD8 T cells, with clustered cells expressing the resident memory markers CD69 and granzyme B but not CD103.


Proinflammatory microenvironments within the intestine regulate the differentiation of tissue-resident CD8⁺ T cells responding to infection.

Bergsbaken T, Bevan MJ - Nat. Immunol. (2015)

Yptb-OVA infection stimulates formation of CD103– CD8 T cell clusters in the LP. C57BL/6 mice received 104 GFP OT-I T cells and were orally infected with Yptb-OVA the following day. The terminal ileum was isolated at 9 days post infection and tissue sections were analyzed by immunohistochemistry. (a) Distribution of OT-I T cells (green) in the ileum. Arrows indicate LP clusters near the crypts and open arrowheads indicate those in the upper part of the villi. Epithelial cells are stained with anti-EpCam antibody (red). Scale bar equals 100 μm. Representative image from 5 mice. (b) Tissue sections were stained with anti-CD103 antibody (red) to determine expression on OT-I T cells (green). Nuclei are stained with DAPI (blue). Open arrowheads indicate CD103+ OT-I T cells. Images of villous (left) and clustered (right) OT-I T cells. (c) OT-I clusters form around areas of Yptb-OVA infection (anti-Yptb-red, OT-I-green, nuclei-blue). (b-c) Representative images from 3 mice. Scale bars equal 25 μm.
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Figure 4: Yptb-OVA infection stimulates formation of CD103– CD8 T cell clusters in the LP. C57BL/6 mice received 104 GFP OT-I T cells and were orally infected with Yptb-OVA the following day. The terminal ileum was isolated at 9 days post infection and tissue sections were analyzed by immunohistochemistry. (a) Distribution of OT-I T cells (green) in the ileum. Arrows indicate LP clusters near the crypts and open arrowheads indicate those in the upper part of the villi. Epithelial cells are stained with anti-EpCam antibody (red). Scale bar equals 100 μm. Representative image from 5 mice. (b) Tissue sections were stained with anti-CD103 antibody (red) to determine expression on OT-I T cells (green). Nuclei are stained with DAPI (blue). Open arrowheads indicate CD103+ OT-I T cells. Images of villous (left) and clustered (right) OT-I T cells. (c) OT-I clusters form around areas of Yptb-OVA infection (anti-Yptb-red, OT-I-green, nuclei-blue). (b-c) Representative images from 3 mice. Scale bars equal 25 μm.
Mentions: Little is known about the distribution of antigen specific CD8 T cells upon colonization with an intestinal pathogen, and we hypothesized the phenotypic differences we observed may be due to different microenvironments created by Yptb infection within the intestine. We examined the localization of GFP OT-I T cells in the intestine of Yptb-OVA infected mice at 9 days post infection. The OT-I cells in the ileum of the small intestine could be divided into two groups based on their distribution: those that were present singly and evenly distributed throughout the tissue, and those that formed clusters (Fig. 4a). These CD8 T cell clusters were found primarily underlying the crypts; however, they could occasionally be found in the villus LP (Fig. 4a). We then examined the surface expression of CD103 on OT-I T cells within the intestine and found that while the majority of the uniformly distributed OT-I cells in IEL and LP expressed CD103 (Fig. 4b left), OT-I T cells in clusters near the crypts were mostly CD103– (Fig. 4b right, Supplementary Fig. 4a). Tissue sections were also stained with Yptb-specific antibodies, indicating OT-I T cells clustered around areas of bacterial infection (Fig. 4c). Similar CD8 T cell clusters were also found in the cecum and colon but not the duodenum (Supplementary Fig. 4b-d). This is consistent with CD8 T cell clusters indicating areas of LP infection, as Yptb infects the distal part of the small intestine, cecum, and colon, but not the proximal small intestine (Supplementary Fig. 4e). These data suggest recruitment to these intestinal aggregates during infection affects the phenotype and function of LP CD8 T cells, with clustered cells expressing the resident memory markers CD69 and granzyme B but not CD103.

Bottom Line: CD103(-) T cells did not require transforming growth factor-β (TGF-β) signaling but were true resident memory cells.Unlike CD103(+)CD8(+) T cells, which were TGF-β dependent and were scattered in the tissue, CD103(-)CD8(+) T cells clustered with CD4(+) T cells and CX3CR1(+) macrophages and/or dendritic cells around areas of bacterial infection.CXCR3-dependent recruitment of cells to inflamed areas was critical for development of the CD103(-) population and pathogen clearance.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Howard Hughes Medical Institute, University of Washington, Seattle, Washington, USA.

ABSTRACT
We report that oral infection with Yersinia pseudotuberculosis results in the development of two distinct populations of pathogen-specific CD8(+) tissue-resident memory T cells (TRM cells) in the lamina propria. CD103(-) T cells did not require transforming growth factor-β (TGF-β) signaling but were true resident memory cells. Unlike CD103(+)CD8(+) T cells, which were TGF-β dependent and were scattered in the tissue, CD103(-)CD8(+) T cells clustered with CD4(+) T cells and CX3CR1(+) macrophages and/or dendritic cells around areas of bacterial infection. CXCR3-dependent recruitment of cells to inflamed areas was critical for development of the CD103(-) population and pathogen clearance. Our studies have identified the 'preferential' development of CD103(-) TRM cells in inflammatory microenvironments within the lamina propria and suggest that this subset has a critical role in controlling infection.

Show MeSH
Related in: MedlinePlus