Journal: Cell metabolism

Article Title: Interleukin 17 drives interstitial entrapment of tissue lipoproteins in experimental psoriasis

doi: 10.1016/j.cmet.2018.10.006

Figure Lengend Snippet: (A) Quantification of insoluble collagen of the heart or the artery from mice that were treated with or without IMQ on the ears for 14 days, or 14 days followed by another 14-day chase; n=5–9 hearts per group. Right panels show trichrome staining of glycolmethacrylate-embedded sections of the carotid artery wall. Scale bar, for both images, is 50 μm. (B) Augmentation pressure was dynamically assessed after surgical placement of a catheter in the carotid artery of mice treated or not with IMQ (with or without chase). N=5–10 mice per condition. (C) The right common carotid artery of PGA1KI/+ mice was photoactivated and plasma fluorescence determined 2 h after photoactivation; some PGA1KI/+ mice were treated with IMQ on both ears for 14 days, with or without another 14 days chase prior to photoactivation. Some groups also received anti-IL-17 neutralizing mAb during the experiment. N=8–14 mice per group. (D-E) apoE−/− mice were treated or not (baseline) with IMQ daily for 3 weeks after having received anti-IL17 neutralizing mAb or isotype control mAb (panel D) or BAPN or vehicle control (panel E). Aortic arch disease was assessed by the percent of en face aorta that was oil red O positive. N=5–10 mice per group. For all panels, data are mean ± SEM. (*P< 0.05; **P<0.01, ***P<0.005).

Article Snippet: DATA AND SOFTWARE AVAILABILITY https://data.mendeley.com/datasets/jp56vt7dcc/1 ADDITIONAL RESOURCES REAGENT or RESOURCE SOURCE IDENTIFIER (RRID) Antibodies Anti-Mouse CD3 (17A2) BD Biosciences AB_2732063 Anti-mouse CD4 (RM4–5) BioLegend AB_2629698 Anti-mouse TCRγ/δ (GL3) BioLegend AB_1731824 Anti-mouse CD19 (6D5) BioLegend AB_313654 Anti-mouse TCRβ (H57–597) BioLegend AB_313429 Anti-mouse CD45.2 (104) BioLegend AB_313445 Anti-mouse IL-17A (TC11–18H10.1) BioLegend AB_961384 Anti-mouse RORγt (Q31–378) BD Biosciences AB_2687545 Anti-Mouse CD4 mAb BioXCell AB_1107636 Anti-Mouse IL17 mAb BioXCell AB_10950102 Anti-mouse IL12/IL23p40 BioXCell AB_1107698 Rat lgG2a isotype control BioXCell AB_1107769 Anti-human ApoA-l EMD Millipore AB_93960 Anti-GFP Abeam AB_303395 Mouse IgA-PE Southern Biotech AB_2732062 Mouse IgM-FITC Sigma AB_2732064 Rabbit anti-Rat Collagen I Cedarlane AB_10060024 Goat Anti-Rabbit IgG HRP Conjugate Bio Rad AB_11125142 Western Diet Harlan Teklad CAT# 88137 Bacterial and Virus Strains AAV8-PGA1 (pCMV) This paper N/A AAV8-PGA1 (pALB) This paper N/A AAV8-D377Y-mPCSK9 Vector Biolabs CAT# AAV-268246 Biological Samples Chemicals, Peptides, and Recombinant Proteins Imiquimod Cream 5% Perrigo.

Techniques: Staining, Fluorescence

Journal: Cell metabolism

Article Title: Interleukin 17 drives interstitial entrapment of tissue lipoproteins in experimental psoriasis

doi: 10.1016/j.cmet.2018.10.006

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: DATA AND SOFTWARE AVAILABILITY https://data.mendeley.com/datasets/jp56vt7dcc/1 ADDITIONAL RESOURCES REAGENT or RESOURCE SOURCE IDENTIFIER (RRID) Antibodies Anti-Mouse CD3 (17A2) BD Biosciences AB_2732063 Anti-mouse CD4 (RM4–5) BioLegend AB_2629698 Anti-mouse TCRγ/δ (GL3) BioLegend AB_1731824 Anti-mouse CD19 (6D5) BioLegend AB_313654 Anti-mouse TCRβ (H57–597) BioLegend AB_313429 Anti-mouse CD45.2 (104) BioLegend AB_313445 Anti-mouse IL-17A (TC11–18H10.1) BioLegend AB_961384 Anti-mouse RORγt (Q31–378) BD Biosciences AB_2687545 Anti-Mouse CD4 mAb BioXCell AB_1107636 Anti-Mouse IL17 mAb BioXCell AB_10950102 Anti-mouse IL12/IL23p40 BioXCell AB_1107698 Rat lgG2a isotype control BioXCell AB_1107769 Anti-human ApoA-l EMD Millipore AB_93960 Anti-GFP Abeam AB_303395 Mouse IgA-PE Southern Biotech AB_2732062 Mouse IgM-FITC Sigma AB_2732064 Rabbit anti-Rat Collagen I Cedarlane AB_10060024 Goat Anti-Rabbit IgG HRP Conjugate Bio Rad AB_11125142 Western Diet Harlan Teklad CAT# 88137 Bacterial and Virus Strains AAV8-PGA1 (pCMV) This paper N/A AAV8-PGA1 (pALB) This paper N/A AAV8-D377Y-mPCSK9 Vector Biolabs CAT# AAV-268246 Biological Samples Chemicals, Peptides, and Recombinant Proteins Imiquimod Cream 5% Perrigo.

Techniques: Western Blot, Virus, Plasmid Preparation, Recombinant, Cream, Labeling, Protease Inhibitor, Staining, Molecular Weight, Marker, Enzyme-linked Immunosorbent Assay, Collagen Assay, Enzymatic Assay, Software

Journal: Frontiers in Immunology

Article Title: Biodegradable nanoparticles induce cGAS/STING-dependent reprogramming of myeloid cells to promote tumor immunotherapy

doi: 10.3389/fimmu.2022.887649

Figure Lengend Snippet: ONP-302 treatment alters immune cell populations within the spleens of B16.F10 tumor-bearing mice over time. Naïve female C57BL/6 mice (n = 9 per treatment group) were injected s.c. with B16.F10 tumor cells. When the tumors were 50-100 mm 3 in size, mice were randomized and treated every three days with saline or ONP-302 (1.0 mg/dose in 200 μL of saline) via i.v. injection. On Day 8 (prior to the first treatment), Day 14 (24 hours after the third treatment), and Day 20 (24 hours after the fifth treatment), spleens were collected to determine the percentage of various immune cell populations. The general lineage markers for CD45 hi , CD4 + T cells, CD8 + T cells, NK cells, CD11b + cells, Ly6C - /Ly6G + , Ly6C + /Ly6G + , and IL-15 cells are presented (A, B) . The specific effector phenotypes of the myeloid (C) , NK (D) , and CD8 + T cells (E) were determined by intracellular staining. The data are presented as the mean percentage of cells ± S.E.M. One representative experiment of two is presented. Asterisks (*, **, ***, ****) indicate a statistically significant difference as compared to saline treated mice, p < 0.05, < 0.01, < 0.001, and < 0.0001 respectively.

Article Snippet: Anti-NK1.1 (clone PK136; BioXCell), anti-IL-15 (clone AIO.3; BioXCell), IgG isotype control antibody (clone C1.18.4; BioXCell) were administered, when indicated, via i.p. injections.

Techniques: Injection, Staining

Journal: Frontiers in Immunology

Article Title: Biodegradable nanoparticles induce cGAS/STING-dependent reprogramming of myeloid cells to promote tumor immunotherapy

doi: 10.3389/fimmu.2022.887649

Figure Lengend Snippet: ONP-302 treatment alters immune cell populations within the tumor microenvironment (TME) of B16.F10 tumor-bearing mice over time. Naïve female C57BL/6 mice (n = 9 per treatment group) were injected s.c. with B16.F10 tumor cells. When the tumors were 50-100 mm 3 in size, mice were randomized and treated every three days with saline or ONP-302 (1.0 mg/dose in 200 μL of saline) via i.v. injection. On Day 8 (prior to the first treatment), Day 14 (24 hours after the third treatment), and Day 20 (24 hours after the fifth treatment), tumors were collected to determine the percentage of various immune cell populations present. The general lineage markers for CD45 hi , CD4 + T cells, CD8 + T cells, NK cells, CD11b + cells, Ly6C - /Ly6G + , Ly6C + /Ly6G + , and IL-15 cells are presented (A, B) . The specific effector phenotypes of the myeloid (C) , NK (D) , and CD8 + T cells (E) were determined by intracellular staining. The data are presented as the mean percentage of cells ± S.E.M. One representative experiment of two is presented. Asterisks (*, **, ***) indicate a statistically significant difference as compared to saline treated mice, p < 0.05, < 0.01, and < 0.001 respectively.

Article Snippet: Anti-NK1.1 (clone PK136; BioXCell), anti-IL-15 (clone AIO.3; BioXCell), IgG isotype control antibody (clone C1.18.4; BioXCell) were administered, when indicated, via i.p. injections.

Techniques: Injection, Staining

Journal: Frontiers in Immunology

Article Title: Biodegradable nanoparticles induce cGAS/STING-dependent reprogramming of myeloid cells to promote tumor immunotherapy

doi: 10.3389/fimmu.2022.887649

Figure Lengend Snippet: Effective ONP-302 treatment requires both adaptive immune and NK cells activated by STING and IL-15, and enhances tumor response to anti-PD-1 immunotherapy. Naïve female C57BL/6, Rag-/- (A) , or Sting-/- (D) mice were injected s.c. was B16.F10 tumor cells. When the tumors were 50-100 mm 3 in size, mice were randomized and treated every three days with saline or ONP-302 (1.0 mg/dose in 200 μL of saline) via i.v. injection. To deplete NK cells, mice were treated with a species and isotype matched control antibody or anti-NK1.1 (100 μg/dose) one day prior to each saline of ONP-302 treatment (n = 5 per treatment group) (B) . To block IL-15, mice were treated with a species and isotype matched control antibody or anti-IL-15 (100 μg/dose) one day prior to each saline of ONP-302 treatment (n=5 per treatment group) (C) . After the fifth dose of ONP-302, mice received three doses of either a control antibody or anti-PD-1 (100 μg/dose) given every three days. The tumor volumes were measured on the indicted days, and the data (E) are presented as the mean tumor volume ± S.E.M. On Day 25 of the disease course, tumors were collected and the percentage of CD45 hi cells from the total cells, the percentage of CD8 + T cells from the CD45 hi cells, and the percentage of CD8 + T cells expressing IFN-γ, CD25, CD44, PD-1 and granzyme was assessed (F) . The data are presented as the mean percentage of the respective parent population of cells. One representative experiment of two is presented. Asterisks (*, **, ***, ****) indicate a statistically significant difference as compared to saline treated mice, p < 0.05, < 0.01, < 0.001, and < 0.0001 respectively.

Article Snippet: Anti-NK1.1 (clone PK136; BioXCell), anti-IL-15 (clone AIO.3; BioXCell), IgG isotype control antibody (clone C1.18.4; BioXCell) were administered, when indicated, via i.p. injections.

Techniques: Injection, Blocking Assay, Expressing

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