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Natural killer T (NKT) cells accelerate Shiga toxin type 2 (Stx2) pathology in mice.

Obata F, Subrahmanyam PB, Vozenilek AE, Hippler LM, Jeffers T, Tongsuk M, Tiper I, Saha P, Jandhyala DM, Kolling GL, Latinovic O, Webb TJ - Front Microbiol (2015)

Bottom Line: NKT cell-associated cytokines such as IL-2, IL-4, IFN-γ, and IL-17 were detected in kidney lysates of Stx2-injected WT mice with the peak around 36 h after Stx2 injection.In CD1KO, there was a delay in the kinetics, and increases in these cytokines were observed 60 h post Stx2 injection.We found that murine glomerular endothelial cells and podocytes express functional CD1d molecules and can present exogenous antigen to NKT cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Maryland School of Medicine Baltimore, MD, USA ; Department of Molecular Pathology, University of Yamanashi Graduate School of Medicine Chuo, Japan.

ABSTRACT
Shiga toxin-producing Escherichia coli (STEC) is a leading cause of childhood renal disease Hemolytic Uremic Syndrome (HUS). The involvement of renal cytokines and chemokines is suspected to play a critical role in disease progression. In current article, we tested the hypothesis that NKT cells are involved in Stx2-induced pathology in vivo. To address this hypothesis we compared Stx2 toxicity in WT and CD1 knockout (KO) mice. In CD1KO mice, which lack natural killer T (NKT) cells, Stx2-induced pathologies such as weight loss, renal failure, and death were delayed. In WT mice, Stx2-specific selective increase in urinary albumin occurs in later time points, and this was also delayed in NKT cell deficient mice. NKT cell-associated cytokines such as IL-2, IL-4, IFN-γ, and IL-17 were detected in kidney lysates of Stx2-injected WT mice with the peak around 36 h after Stx2 injection. In CD1KO, there was a delay in the kinetics, and increases in these cytokines were observed 60 h post Stx2 injection. These data suggest that NKT cells accelerate Stx2-induced pathology in mouse kidneys. To determine the mechanism by which NKT cells promote Stx2-associated disease, in vitro studies were performed using murine renal cells. We found that murine glomerular endothelial cells and podocytes express functional CD1d molecules and can present exogenous antigen to NKT cells. Moreover, we observed the direct interaction between Stx2 and the receptor Gb3 on the surface of mouse renal cells by 3D STORM-TIRF which provides single molecule imaging. Collectively, these data suggest that Stx2 binds to Gb3 on renal cells and leads to aberrant CD1d-mediated NKT cell activation. Therefore, strategies targeting NKT cells could have a significant impact on Stx2-associated renal pathology in STEC disease.

No MeSH data available.


Related in: MedlinePlus

CD1KO mice show delayed renal cytokine release following treatment with Stx2. (A–D) WT (n = 3 per time point) or CD1KO (n = 3 per time point) mice were treated with Stx2 for the indicated time periods. Kidney lysates from both treatment groups at the indicated time points were analyzed by ELISA and (A) IL-2, (B) IL-4, (C) IFN-γ and (D) IL-17 concentrations were determined. Results were normalized to total protein and have been represented as pg of cytokine per mg total protein. One-Way ANOVA followed by Tukey test, *p < 0.001 vs. WT 0 h, $p < 0.001 vs. WT 36 h, @p < 0.05 vs. WT 36 h and #p < 0.001 vs. CD1KO 0 h.
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Figure 2: CD1KO mice show delayed renal cytokine release following treatment with Stx2. (A–D) WT (n = 3 per time point) or CD1KO (n = 3 per time point) mice were treated with Stx2 for the indicated time periods. Kidney lysates from both treatment groups at the indicated time points were analyzed by ELISA and (A) IL-2, (B) IL-4, (C) IFN-γ and (D) IL-17 concentrations were determined. Results were normalized to total protein and have been represented as pg of cytokine per mg total protein. One-Way ANOVA followed by Tukey test, *p < 0.001 vs. WT 0 h, $p < 0.001 vs. WT 36 h, @p < 0.05 vs. WT 36 h and #p < 0.001 vs. CD1KO 0 h.

Mentions: Given the delay in Stx2-pathologies in CD1KO mice, we suspected the induction of NKT cell-associated inflammation in kidney. NKT cells produce cytokines such as IL-2, IL-4, IFN-γ, and IL-17. Kidney lysates of Stx2-injected mice were analyzed for these cytokines (Figure 2). In WT mice, IL-2, IL-4, IFN-γ, and IL-17 showed increase after 8 h and peaked at 36–48 h following Stx2 injection. In line with previous markers of Stx2-mediated pathology in CD1KO mice, significant increases in these cytokines were noted later at 60 h. These data suggest that NKT cell activation contributes to accelerate Stx2 renal pathology by promoting inflammation.


Natural killer T (NKT) cells accelerate Shiga toxin type 2 (Stx2) pathology in mice.

Obata F, Subrahmanyam PB, Vozenilek AE, Hippler LM, Jeffers T, Tongsuk M, Tiper I, Saha P, Jandhyala DM, Kolling GL, Latinovic O, Webb TJ - Front Microbiol (2015)

CD1KO mice show delayed renal cytokine release following treatment with Stx2. (A–D) WT (n = 3 per time point) or CD1KO (n = 3 per time point) mice were treated with Stx2 for the indicated time periods. Kidney lysates from both treatment groups at the indicated time points were analyzed by ELISA and (A) IL-2, (B) IL-4, (C) IFN-γ and (D) IL-17 concentrations were determined. Results were normalized to total protein and have been represented as pg of cytokine per mg total protein. One-Way ANOVA followed by Tukey test, *p < 0.001 vs. WT 0 h, $p < 0.001 vs. WT 36 h, @p < 0.05 vs. WT 36 h and #p < 0.001 vs. CD1KO 0 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4389548&req=5

Figure 2: CD1KO mice show delayed renal cytokine release following treatment with Stx2. (A–D) WT (n = 3 per time point) or CD1KO (n = 3 per time point) mice were treated with Stx2 for the indicated time periods. Kidney lysates from both treatment groups at the indicated time points were analyzed by ELISA and (A) IL-2, (B) IL-4, (C) IFN-γ and (D) IL-17 concentrations were determined. Results were normalized to total protein and have been represented as pg of cytokine per mg total protein. One-Way ANOVA followed by Tukey test, *p < 0.001 vs. WT 0 h, $p < 0.001 vs. WT 36 h, @p < 0.05 vs. WT 36 h and #p < 0.001 vs. CD1KO 0 h.
Mentions: Given the delay in Stx2-pathologies in CD1KO mice, we suspected the induction of NKT cell-associated inflammation in kidney. NKT cells produce cytokines such as IL-2, IL-4, IFN-γ, and IL-17. Kidney lysates of Stx2-injected mice were analyzed for these cytokines (Figure 2). In WT mice, IL-2, IL-4, IFN-γ, and IL-17 showed increase after 8 h and peaked at 36–48 h following Stx2 injection. In line with previous markers of Stx2-mediated pathology in CD1KO mice, significant increases in these cytokines were noted later at 60 h. These data suggest that NKT cell activation contributes to accelerate Stx2 renal pathology by promoting inflammation.

Bottom Line: NKT cell-associated cytokines such as IL-2, IL-4, IFN-γ, and IL-17 were detected in kidney lysates of Stx2-injected WT mice with the peak around 36 h after Stx2 injection.In CD1KO, there was a delay in the kinetics, and increases in these cytokines were observed 60 h post Stx2 injection.We found that murine glomerular endothelial cells and podocytes express functional CD1d molecules and can present exogenous antigen to NKT cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Maryland School of Medicine Baltimore, MD, USA ; Department of Molecular Pathology, University of Yamanashi Graduate School of Medicine Chuo, Japan.

ABSTRACT
Shiga toxin-producing Escherichia coli (STEC) is a leading cause of childhood renal disease Hemolytic Uremic Syndrome (HUS). The involvement of renal cytokines and chemokines is suspected to play a critical role in disease progression. In current article, we tested the hypothesis that NKT cells are involved in Stx2-induced pathology in vivo. To address this hypothesis we compared Stx2 toxicity in WT and CD1 knockout (KO) mice. In CD1KO mice, which lack natural killer T (NKT) cells, Stx2-induced pathologies such as weight loss, renal failure, and death were delayed. In WT mice, Stx2-specific selective increase in urinary albumin occurs in later time points, and this was also delayed in NKT cell deficient mice. NKT cell-associated cytokines such as IL-2, IL-4, IFN-γ, and IL-17 were detected in kidney lysates of Stx2-injected WT mice with the peak around 36 h after Stx2 injection. In CD1KO, there was a delay in the kinetics, and increases in these cytokines were observed 60 h post Stx2 injection. These data suggest that NKT cells accelerate Stx2-induced pathology in mouse kidneys. To determine the mechanism by which NKT cells promote Stx2-associated disease, in vitro studies were performed using murine renal cells. We found that murine glomerular endothelial cells and podocytes express functional CD1d molecules and can present exogenous antigen to NKT cells. Moreover, we observed the direct interaction between Stx2 and the receptor Gb3 on the surface of mouse renal cells by 3D STORM-TIRF which provides single molecule imaging. Collectively, these data suggest that Stx2 binds to Gb3 on renal cells and leads to aberrant CD1d-mediated NKT cell activation. Therefore, strategies targeting NKT cells could have a significant impact on Stx2-associated renal pathology in STEC disease.

No MeSH data available.


Related in: MedlinePlus