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Local induction of immunosuppressive CD8+ T cells in the gut-associated lymphoid tissues.

Fleissner D, Hansen W, Geffers R, Buer J, Westendorf AM - PLoS ONE (2010)

Bottom Line: HA-specific CD8(+) T cells were isolated from gut-associated tissues and phenotypically and functionally characterized for the expression of Foxp3(+) and their suppressive capacity.Finally, gut-associated dendritic cells were able to induce HA-specific CD8(+)Foxp3(+) T cells.We demonstrate that gut specific antigen presentation is sufficient to induce CD8(+) T(regs)in vivo which may maintain intestinal homeostasis by down-modulating effector functions of T cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Medical Microbiology, University Hospital Essen, University Duisburg-Essen, Essen, Germany.

ABSTRACT

Background: In contrast to intestinal CD4(+) regulatory T cells (T(regs)), the generation and function of immunomodulatory intestinal CD8(+) T cells is less well defined. To dissect the immunologic mechanisms of CD8(+) T cell function in the mucosa, reactivity against hemagglutinin (HA) expressed in intestinal epithelial cells of mice bearing a MHC class-I-restricted T-cell-receptor specific for HA was studied.

Methodology and principal findings: HA-specific CD8(+) T cells were isolated from gut-associated tissues and phenotypically and functionally characterized for the expression of Foxp3(+) and their suppressive capacity. We demonstrate that intestinal HA expression led to peripheral induction of HA-specific CD8(+)Foxp3(+) T cells. Antigen-experienced CD8(+) T cells in this transgenic mouse model suppressed the proliferation of CD8(+) and CD4(+) T cells in vitro. Gene expression analysis of suppressive HA-specific CD8(+) T cells revealed a specific up-regulation of CD103, Nrp1, Tnfrsf9 and Pdcd1, molecules also expressed on CD4(+) T(reg) subsets. Finally, gut-associated dendritic cells were able to induce HA-specific CD8(+)Foxp3(+) T cells.

Conclusion and significance: We demonstrate that gut specific antigen presentation is sufficient to induce CD8(+) T(regs)in vivo which may maintain intestinal homeostasis by down-modulating effector functions of T cells.

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HA-specific CD8+ T cells from VILLIN-HA/CL4-TCR exhibit suppressive capacity.HA-specific CD8+ T cells were isolated from the MLN of CL4-TCR and VILLIN-HA/CL4-TCR transgenic mice by cell sorting and co-cultured with CFSE-labeled HA-specific CD4+ or CD8+ responder cells in the presence of the cognate peptide. Proliferation of responder cells was measured by loss of CFSE dye. Data shown are representative of three independent experiments.
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pone-0015373-g002: HA-specific CD8+ T cells from VILLIN-HA/CL4-TCR exhibit suppressive capacity.HA-specific CD8+ T cells were isolated from the MLN of CL4-TCR and VILLIN-HA/CL4-TCR transgenic mice by cell sorting and co-cultured with CFSE-labeled HA-specific CD4+ or CD8+ responder cells in the presence of the cognate peptide. Proliferation of responder cells was measured by loss of CFSE dye. Data shown are representative of three independent experiments.

Mentions: We recently demonstrated that in VILLIN-HA/CL4-TCR transgenic mice chronic intestinal antigen exposure leads to the infiltration of HA-specific CD8+ T cells into the intestinal mucosa but without development of severe intestinal inflammation [10]. In the present study we further analyzed the infiltrating CD8+ T cells and demonstrated a partial induction of CD8+Foxp3+ T cells which was restricted to the periphery (Sp and MLN) and not detectable in the thymus (Figure 1). Characterization of HA-specific CD8+ T cells isolated from the MLN of VILLIN-HA/CL4-TCR transgenic mice reflected a reduced secretion of IFN-γ, TNF-α, and IL-6 after in vitro stimulation (Table 1). To analyze these antigen experienced HA-specific CD8+ T cells of VILLIN-HA/CL4-TCR transgenic mice in more detail the suppressive capacity in vitro was measured. For this purpose HA-specific CD8+ T cells were FACS-sorted by pentamer-staining (H-2KdIYSTVASSL+) from the MLN of control CL4-TCR and VILLIN-HA/CL4-TCR transgenic mice and co-cultured with naïve CFSE-labeled HA-specific CD8+ or CD4+ responder T cells in the presence of APCs and cognate HA peptide. Interestingly, HA-specific CD8+ T cells isolated from VILLIN-HA/CL4-TCR transgenic mice were able to suppress both CD8+ and CD4+ T cell proliferation in vitro (Figure 2).


Local induction of immunosuppressive CD8+ T cells in the gut-associated lymphoid tissues.

Fleissner D, Hansen W, Geffers R, Buer J, Westendorf AM - PLoS ONE (2010)

HA-specific CD8+ T cells from VILLIN-HA/CL4-TCR exhibit suppressive capacity.HA-specific CD8+ T cells were isolated from the MLN of CL4-TCR and VILLIN-HA/CL4-TCR transgenic mice by cell sorting and co-cultured with CFSE-labeled HA-specific CD4+ or CD8+ responder cells in the presence of the cognate peptide. Proliferation of responder cells was measured by loss of CFSE dye. Data shown are representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2958146&req=5

pone-0015373-g002: HA-specific CD8+ T cells from VILLIN-HA/CL4-TCR exhibit suppressive capacity.HA-specific CD8+ T cells were isolated from the MLN of CL4-TCR and VILLIN-HA/CL4-TCR transgenic mice by cell sorting and co-cultured with CFSE-labeled HA-specific CD4+ or CD8+ responder cells in the presence of the cognate peptide. Proliferation of responder cells was measured by loss of CFSE dye. Data shown are representative of three independent experiments.
Mentions: We recently demonstrated that in VILLIN-HA/CL4-TCR transgenic mice chronic intestinal antigen exposure leads to the infiltration of HA-specific CD8+ T cells into the intestinal mucosa but without development of severe intestinal inflammation [10]. In the present study we further analyzed the infiltrating CD8+ T cells and demonstrated a partial induction of CD8+Foxp3+ T cells which was restricted to the periphery (Sp and MLN) and not detectable in the thymus (Figure 1). Characterization of HA-specific CD8+ T cells isolated from the MLN of VILLIN-HA/CL4-TCR transgenic mice reflected a reduced secretion of IFN-γ, TNF-α, and IL-6 after in vitro stimulation (Table 1). To analyze these antigen experienced HA-specific CD8+ T cells of VILLIN-HA/CL4-TCR transgenic mice in more detail the suppressive capacity in vitro was measured. For this purpose HA-specific CD8+ T cells were FACS-sorted by pentamer-staining (H-2KdIYSTVASSL+) from the MLN of control CL4-TCR and VILLIN-HA/CL4-TCR transgenic mice and co-cultured with naïve CFSE-labeled HA-specific CD8+ or CD4+ responder T cells in the presence of APCs and cognate HA peptide. Interestingly, HA-specific CD8+ T cells isolated from VILLIN-HA/CL4-TCR transgenic mice were able to suppress both CD8+ and CD4+ T cell proliferation in vitro (Figure 2).

Bottom Line: HA-specific CD8(+) T cells were isolated from gut-associated tissues and phenotypically and functionally characterized for the expression of Foxp3(+) and their suppressive capacity.Finally, gut-associated dendritic cells were able to induce HA-specific CD8(+)Foxp3(+) T cells.We demonstrate that gut specific antigen presentation is sufficient to induce CD8(+) T(regs)in vivo which may maintain intestinal homeostasis by down-modulating effector functions of T cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Medical Microbiology, University Hospital Essen, University Duisburg-Essen, Essen, Germany.

ABSTRACT

Background: In contrast to intestinal CD4(+) regulatory T cells (T(regs)), the generation and function of immunomodulatory intestinal CD8(+) T cells is less well defined. To dissect the immunologic mechanisms of CD8(+) T cell function in the mucosa, reactivity against hemagglutinin (HA) expressed in intestinal epithelial cells of mice bearing a MHC class-I-restricted T-cell-receptor specific for HA was studied.

Methodology and principal findings: HA-specific CD8(+) T cells were isolated from gut-associated tissues and phenotypically and functionally characterized for the expression of Foxp3(+) and their suppressive capacity. We demonstrate that intestinal HA expression led to peripheral induction of HA-specific CD8(+)Foxp3(+) T cells. Antigen-experienced CD8(+) T cells in this transgenic mouse model suppressed the proliferation of CD8(+) and CD4(+) T cells in vitro. Gene expression analysis of suppressive HA-specific CD8(+) T cells revealed a specific up-regulation of CD103, Nrp1, Tnfrsf9 and Pdcd1, molecules also expressed on CD4(+) T(reg) subsets. Finally, gut-associated dendritic cells were able to induce HA-specific CD8(+)Foxp3(+) T cells.

Conclusion and significance: We demonstrate that gut specific antigen presentation is sufficient to induce CD8(+) T(regs)in vivo which may maintain intestinal homeostasis by down-modulating effector functions of T cells.

Show MeSH