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T cell-intrinsic role of Nod2 in promoting type 1 immunity to Toxoplasma gondii.

Shaw MH, Reimer T, Sánchez-Valdepeñas C, Warner N, Kim YG, Fresno M, Nuñez G - Nat. Immunol. (2009)

Bottom Line: Nod2 deficiency results in an impaired immune response to bacterial pathogens.Nod2(-/-) CD4(+) T cells had poor helper T cell differentiation, which was associated with impaired production of interleukin 2 (IL-2) and nuclear accumulation of the transcription factor subunit c-Rel.Our data demonstrate a T cell-intrinsic role for Nod2 signaling that is critical for host defense against T. gondii.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Comprehensive Cancer Center, University of Michigan Medical School, Ann Arbor, Michigan, USA.

ABSTRACT
Nod2 belongs to the nucleotide-binding oligomerization domain receptor (NLR) family of proteins, which function as intracellular pathogen sensors in innate immune cells. Nod2 deficiency results in an impaired immune response to bacterial pathogens. However, how this protein promotes host defense against intracellular parasites is unknown. Here we found that Nod2(-/-) mice had less clearance of Toxoplasma gondii and lower interferon-gamma (IFN-gamma) production. Reconstitution of T cell-deficient mice with Nod2(-/-) T cells followed by T. gondii infection demonstrated a T cell-intrinsic defect. Nod2(-/-) CD4(+) T cells had poor helper T cell differentiation, which was associated with impaired production of interleukin 2 (IL-2) and nuclear accumulation of the transcription factor subunit c-Rel. Our data demonstrate a T cell-intrinsic role for Nod2 signaling that is critical for host defense against T. gondii.

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Nod2−/− CD4 cells have an intrinsic defect in IFN-γ production. (a) Left, MACS-sorted splenic CD4+ cells from wild-type (Thy-1.1) and Nod2−/− (Thy-1.1) mice were injected intravenously into wild-type Tcrb−/− (Thy-1.2) or Nod2−/−Tcrb−/− (Thy-1.2) recipients, which were then challenged with irradiated CPS parasites. On day 9 post challenge, PECs were harvested from reconstituted mice and IFN-γ production was determined by intracellular cytokine staining following in vitro stimulation with live parasite. Each dot plots is representative of 4–8 mice per group and is gated on CD4+TCR-β+Thy-1.1+ cells. Right, the data were pooled; each dot represents one mouse and horizontal bars represent the mean. (b) PECs of reconstituted mice as described in (a) were co-cultured with live parasite for 24 h and IFN-γ in the supernatants was determined by ELISA.
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Figure 5: Nod2−/− CD4 cells have an intrinsic defect in IFN-γ production. (a) Left, MACS-sorted splenic CD4+ cells from wild-type (Thy-1.1) and Nod2−/− (Thy-1.1) mice were injected intravenously into wild-type Tcrb−/− (Thy-1.2) or Nod2−/−Tcrb−/− (Thy-1.2) recipients, which were then challenged with irradiated CPS parasites. On day 9 post challenge, PECs were harvested from reconstituted mice and IFN-γ production was determined by intracellular cytokine staining following in vitro stimulation with live parasite. Each dot plots is representative of 4–8 mice per group and is gated on CD4+TCR-β+Thy-1.1+ cells. Right, the data were pooled; each dot represents one mouse and horizontal bars represent the mean. (b) PECs of reconstituted mice as described in (a) were co-cultured with live parasite for 24 h and IFN-γ in the supernatants was determined by ELISA.

Mentions: To directly assess the functional role of Nod2 in T cells, wild-type and Nod2−/− CD4+ T cells were adoptively transferred into Tcrb−/− or Tcrb−/−Nod2−/− recipients. The reconstituted mice were then immunized with CPS and the frequency of effector T cells was assessed 9 days later. Tcrb−/− and Tcrb−/−Nod2−/− recipient mice reconstituted with wild-type CD4+ T cells exhibit high frequencies of IFN-γ+ CD4+ T lymphocytes following parasite challenge (Figure 5a). However, there was a significant reduction in the percentage of IFN-γ+ CD4+ T lymphocytes in mice reconstituted with Nod2-deficient CD4+ T cells regardless of the genotype of the recipient mice (Figure 5a). The impaired CD4+ T cell response is consistent with the lower IFN-γ production by PECs harvested from Tcrb−/− and Tcrb−/−Nod2−/− mice reconstituted with Nod2−/− CD4+ T cells (Figure 5b). Lastly, the resistance of Nod2−/− mice following T. gondii infection was enhanced by the presence of wild-type T cells (Supplementary Figure 4). Taken together, these findings indicate that the impaired T cell response observed in Nod2−/− mice during infection is associated with an intrinsic defect in Nod2-deficient T cells.


T cell-intrinsic role of Nod2 in promoting type 1 immunity to Toxoplasma gondii.

Shaw MH, Reimer T, Sánchez-Valdepeñas C, Warner N, Kim YG, Fresno M, Nuñez G - Nat. Immunol. (2009)

Nod2−/− CD4 cells have an intrinsic defect in IFN-γ production. (a) Left, MACS-sorted splenic CD4+ cells from wild-type (Thy-1.1) and Nod2−/− (Thy-1.1) mice were injected intravenously into wild-type Tcrb−/− (Thy-1.2) or Nod2−/−Tcrb−/− (Thy-1.2) recipients, which were then challenged with irradiated CPS parasites. On day 9 post challenge, PECs were harvested from reconstituted mice and IFN-γ production was determined by intracellular cytokine staining following in vitro stimulation with live parasite. Each dot plots is representative of 4–8 mice per group and is gated on CD4+TCR-β+Thy-1.1+ cells. Right, the data were pooled; each dot represents one mouse and horizontal bars represent the mean. (b) PECs of reconstituted mice as described in (a) were co-cultured with live parasite for 24 h and IFN-γ in the supernatants was determined by ELISA.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2803073&req=5

Figure 5: Nod2−/− CD4 cells have an intrinsic defect in IFN-γ production. (a) Left, MACS-sorted splenic CD4+ cells from wild-type (Thy-1.1) and Nod2−/− (Thy-1.1) mice were injected intravenously into wild-type Tcrb−/− (Thy-1.2) or Nod2−/−Tcrb−/− (Thy-1.2) recipients, which were then challenged with irradiated CPS parasites. On day 9 post challenge, PECs were harvested from reconstituted mice and IFN-γ production was determined by intracellular cytokine staining following in vitro stimulation with live parasite. Each dot plots is representative of 4–8 mice per group and is gated on CD4+TCR-β+Thy-1.1+ cells. Right, the data were pooled; each dot represents one mouse and horizontal bars represent the mean. (b) PECs of reconstituted mice as described in (a) were co-cultured with live parasite for 24 h and IFN-γ in the supernatants was determined by ELISA.
Mentions: To directly assess the functional role of Nod2 in T cells, wild-type and Nod2−/− CD4+ T cells were adoptively transferred into Tcrb−/− or Tcrb−/−Nod2−/− recipients. The reconstituted mice were then immunized with CPS and the frequency of effector T cells was assessed 9 days later. Tcrb−/− and Tcrb−/−Nod2−/− recipient mice reconstituted with wild-type CD4+ T cells exhibit high frequencies of IFN-γ+ CD4+ T lymphocytes following parasite challenge (Figure 5a). However, there was a significant reduction in the percentage of IFN-γ+ CD4+ T lymphocytes in mice reconstituted with Nod2-deficient CD4+ T cells regardless of the genotype of the recipient mice (Figure 5a). The impaired CD4+ T cell response is consistent with the lower IFN-γ production by PECs harvested from Tcrb−/− and Tcrb−/−Nod2−/− mice reconstituted with Nod2−/− CD4+ T cells (Figure 5b). Lastly, the resistance of Nod2−/− mice following T. gondii infection was enhanced by the presence of wild-type T cells (Supplementary Figure 4). Taken together, these findings indicate that the impaired T cell response observed in Nod2−/− mice during infection is associated with an intrinsic defect in Nod2-deficient T cells.

Bottom Line: Nod2 deficiency results in an impaired immune response to bacterial pathogens.Nod2(-/-) CD4(+) T cells had poor helper T cell differentiation, which was associated with impaired production of interleukin 2 (IL-2) and nuclear accumulation of the transcription factor subunit c-Rel.Our data demonstrate a T cell-intrinsic role for Nod2 signaling that is critical for host defense against T. gondii.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Comprehensive Cancer Center, University of Michigan Medical School, Ann Arbor, Michigan, USA.

ABSTRACT
Nod2 belongs to the nucleotide-binding oligomerization domain receptor (NLR) family of proteins, which function as intracellular pathogen sensors in innate immune cells. Nod2 deficiency results in an impaired immune response to bacterial pathogens. However, how this protein promotes host defense against intracellular parasites is unknown. Here we found that Nod2(-/-) mice had less clearance of Toxoplasma gondii and lower interferon-gamma (IFN-gamma) production. Reconstitution of T cell-deficient mice with Nod2(-/-) T cells followed by T. gondii infection demonstrated a T cell-intrinsic defect. Nod2(-/-) CD4(+) T cells had poor helper T cell differentiation, which was associated with impaired production of interleukin 2 (IL-2) and nuclear accumulation of the transcription factor subunit c-Rel. Our data demonstrate a T cell-intrinsic role for Nod2 signaling that is critical for host defense against T. gondii.

Show MeSH
Related in: MedlinePlus