Journal: PLoS Pathogens

Article Title: Foxp3 + Regulatory T Cells Delay Expulsion of Intestinal Nematodes by Suppression of IL-9-Driven Mast Cell Activation in BALB/c but Not in C57BL/6 Mice

doi: 10.1371/journal.ppat.1003913

Figure Lengend Snippet: BALB/c (white bars), BALB/c DEREG (black bars), C57BL/6 (light grey bars), and C57BL/6 DEREG (dark grey bars) mice were treated with DT on three consecutive days starting one day before S. ratti infection. A : Polyclonal IgE in the serum was quantified at the indicated time points. Shown is one experiment (n = 6 per time point) that is representative for one independent repeat. Asterisks indicate significant difference of the mean analyzed by one-way ANOVA with Bonferroni post test (*** p≤0.001). B: S. ratti -specific IgM was quantified in the serum on day 7 p.i. Shown are the combined results of three independent experiments (n = 9–11). Asterisks indicate significant difference of the mean analyzed by students t test (* p≤0.05). C–G: Mice were sacrificed on day 6 p.i. and splenocytes (2×10 5 ) were stimulated for 72 h with S. ratti lysate (upper panel) or with α-CD3 (lower panel). IL-13 ( C ), IL-4 ( D ), IL-10 ( E ), IL-3 ( F ) and IL-9 ( G ) in the supernatant were quantified by ELISA. Splenocytes that were cultivated in the presence of medium only did not secrete cytokines (data not shown). Shown are the combined results of two independent experiments (n = 8) for C–F and combined results of four independent experiments (n = 14–16) for G . Asterisks indicate significant difference of the mean analyzed by one-way ANOVA with Bonferroni post test (* p≤0.05, ** p≤0.01, *** p≤0.001).

Article Snippet: Samples and recombinant IL-9 standard (Peprotech, Hamburg, Germany) were incubated overnight and detection was performed with an anti-IL-9-biotin AB (BD, Heidelberg, Germany) for 1 h at RT and subsequent Streptavidin-HRP incubation for 20 min before development with 100 µL/well tetramethylbenzidine 0.1 mg/ml, 0.003% H 2 O 2 in 100 mM NaH 2 PO 4 (pH 5.5).

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: PLoS Pathogens

Article Title: Foxp3 + Regulatory T Cells Delay Expulsion of Intestinal Nematodes by Suppression of IL-9-Driven Mast Cell Activation in BALB/c but Not in C57BL/6 Mice

doi: 10.1371/journal.ppat.1003913

Figure Lengend Snippet: A: Experimental setup for IL-9 treatment is shown. B–C: Number of parasitic adults in the small intestine of infected non-treated BALB/c ( B , white) and C57BL/6 ( C , light grey) mice or infected and IL-9 treated BALB/c ( B , black) and C57BL/6 ( C , dark grey) mice on day 6 p.i. Shown are the combined results of three independent experiments (n = 12). D: Experimental setup for α-IL-9 treatment is shown. E–F: Number of parasitic adults in the small intestine of infected isotype treated BALB/c ( D , white) and C57BL/6 ( E , light grey) mice or infected and α-IL-9 treated BALB/c ( D , black) and C57BL/6 ( E , dark grey) mice on day 6 p.i. G–H : Concentration of MMCP-1 in the serum of infected isotype treated or α-IL-9 treated BALB/c ( G ) and C57BL/6 ( H ) mice on day 4 and day 6 p.i. Shown are the combined results of two independent experiments (n = 8). Asterisks indicate significant differences of the mean analyzed by students t test (** p≤0.01, *** p≤0.001).

Article Snippet: Samples and recombinant IL-9 standard (Peprotech, Hamburg, Germany) were incubated overnight and detection was performed with an anti-IL-9-biotin AB (BD, Heidelberg, Germany) for 1 h at RT and subsequent Streptavidin-HRP incubation for 20 min before development with 100 µL/well tetramethylbenzidine 0.1 mg/ml, 0.003% H 2 O 2 in 100 mM NaH 2 PO 4 (pH 5.5).

Techniques: Infection, Concentration Assay

Journal: PLoS Pathogens

Article Title: Foxp3 + Regulatory T Cells Delay Expulsion of Intestinal Nematodes by Suppression of IL-9-Driven Mast Cell Activation in BALB/c but Not in C57BL/6 Mice

doi: 10.1371/journal.ppat.1003913

Figure Lengend Snippet: A: Experimental setup is shown. B: Number of parasitic adults in the small intestine of BALB/c (white), Treg-depleted BALB/c DEREG (black), α-IL-9 treated BALB/c (light blue) and α-IL-9 treated Treg-depleted BALB/c DEREG (dark blue) mice on day 6 p.i. C: Number of parasitic adults in the small intestine of BALB/c (white), Treg-depleted BALB/c DEREG (black), α-IL-13 treated BALB/c (light green) and α-IL-13 treated Treg-depleted BALB/c DEREG (dark green) mice on day 6 p.i. D: Concentration of MMCP-1 in the serum of infected mice at indicated time points. Shown are the combined results of four ( BD ) or two ( C ) independent experiments (BD: n = 14, C: n = 8). Asterisks indicate significant difference of the mean analyzed by ( BC ) students t test or ( D ) one-way ANOVA with Bonferroni post test (*** p≤0.001).

Article Snippet: Samples and recombinant IL-9 standard (Peprotech, Hamburg, Germany) were incubated overnight and detection was performed with an anti-IL-9-biotin AB (BD, Heidelberg, Germany) for 1 h at RT and subsequent Streptavidin-HRP incubation for 20 min before development with 100 µL/well tetramethylbenzidine 0.1 mg/ml, 0.003% H 2 O 2 in 100 mM NaH 2 PO 4 (pH 5.5).

Techniques: Concentration Assay, Infection

Journal: PLoS Pathogens

Article Title: Foxp3 + Regulatory T Cells Delay Expulsion of Intestinal Nematodes by Suppression of IL-9-Driven Mast Cell Activation in BALB/c but Not in C57BL/6 Mice

doi: 10.1371/journal.ppat.1003913

Figure Lengend Snippet: A: Experimental setup for α-IL-9 treatment either early (blue arrows) or late (grey arrows) in S. ratti infection is shown. B: Graph shows number of parasitic adults in the small intestine of BALB/c (white), Treg-depleted, isotype treated BALB/c DEREG (black), Treg-depleted early α-IL-9 treated BALB/c DEREG (dark blue) and Treg-depleted late α-IL-9 treated BALB/c DEREG (dark grey) mice on day 6 p.i. Shown are the combined results of three independent experiments (n = 14). C: Concentrations of MMCP-1 in the serum of infected mice on day 4 and day 6 p.i. are shown as combined results of two independent experiments (n = 9). Asterisks indicate significant difference of the mean analyzed by one-way ANOVA with Bonferroni post test (* p≤0.05, *** p≤0.001).

Article Snippet: Samples and recombinant IL-9 standard (Peprotech, Hamburg, Germany) were incubated overnight and detection was performed with an anti-IL-9-biotin AB (BD, Heidelberg, Germany) for 1 h at RT and subsequent Streptavidin-HRP incubation for 20 min before development with 100 µL/well tetramethylbenzidine 0.1 mg/ml, 0.003% H 2 O 2 in 100 mM NaH 2 PO 4 (pH 5.5).

Techniques: Infection

Journal: PLoS Pathogens

Article Title: Foxp3 + Regulatory T Cells Delay Expulsion of Intestinal Nematodes by Suppression of IL-9-Driven Mast Cell Activation in BALB/c but Not in C57BL/6 Mice

doi: 10.1371/journal.ppat.1003913

Figure Lengend Snippet: AB: BALB/c Cpa3 WT (white bars), BALB/c DEREG Cpa3 WT (black bars), BALB/c Cpa3 CRE (light green bars) and BALB/c DEREG Cpa3 CRE (dark green bars) mice were treated with DT on three consecutive days starting one day before s.c infection with 2000 S. ratti iL3. A: Number of parasitic adults in the small intestine was counted on day 6 p.i. Depicted are the combined results of three independent experiments (n≥12) and error bars show SEM. B: Splenocytes were prepared at day 6 p.i. and cultured in the presence of α-CD3 for 72 h. IL-9 concentration in the supernatant was quantified by ELISA. Unstimulated splenocytes did not secrete detectable IL-9. Shown are the combined results of three independent experiments (n≥12). AB: Asterisks indicate significant difference of the mean analyzed by students t test (*** p≤0.001).

Article Snippet: Samples and recombinant IL-9 standard (Peprotech, Hamburg, Germany) were incubated overnight and detection was performed with an anti-IL-9-biotin AB (BD, Heidelberg, Germany) for 1 h at RT and subsequent Streptavidin-HRP incubation for 20 min before development with 100 µL/well tetramethylbenzidine 0.1 mg/ml, 0.003% H 2 O 2 in 100 mM NaH 2 PO 4 (pH 5.5).

Techniques: Infection, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay