Journal: Scientific Reports

Article Title: T. gondii infection induces IL-1R dependent chronic cachexia and perivascular fibrosis in the liver and skeletal muscle

doi: 10.1038/s41598-020-72767-0

Figure Lengend Snippet: Chronic Toxoplasma infection causes sustained cachexia in mice. 10–14 week old C57BL/6J mice were intraperitoneally infected with 10 Me49-GFP-luciferase T. gondii cysts (I, red) or mock injected with PBS (UI, black). ( a ) Schematic of weight loss relative to parasite distribution. The acute phase of infection (white) is dominated by Toxoplasma tachyzoites (green crescents) which spread systemically, infecting most tissues in the body. 4–6 weeks post-infection (wpi), systemic infection is largely cleared and parasites are driven to the chronic tissue cyst form (green circles). ( b ) Mice lose up to 20% of their initial body mass in the first 4 wpi and fail to regain weight relative to uninfected controls. N = 35–45 mice pooled from 3 independent experiments. ( c ) Dolichos biflorous positive T. gondii cysts per half brain at 5–9 wpi. N = 19 pooled from 6 independent experiments. ( d ) Daily food intake per cage normalized to pooled weight of the mice in the cage measured every 24 h. N = 7–9 cages per group, pooled from 3 independent experiments. ( e ) Mice were individually housed for 24 h at 10 wpi and food intake over 24 h was determined by weight (left) and caloric content of fecal pellets were determined by bomb calorimetry (right). N = 4 mice per group. ( f ) Echo MRI quantification of fat (left) and lean (right) tissue mass at 2 or 6 wpi. N = 28–45 mice per group, representative of 3 independent experiments. ( g ) Inguinal subcutaneous white adipose tissue (scWAT), epididymal visceral white adipose tissue (vWAT), quadriceps (Quad) and liver weights at 2 wpi or 9 wpi. N = 12–18 mice per group, pooled from 3 experiments. ( h ) Quantity of T. gondii DNA relative to host beta-actin in the tibialis anterior muscle at 9 wpi. N = 4–5 mice per group. n.d. not detectable. ( i ) Serum cytokines measured by Luminex at 1 or 5 wpi. N = 3–4 mice per group, representative of 2 experiments. Error bars are standard error of the mean. *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001 by unpaired Student’s t test.

Article Snippet: 1 × 10 4 cells were seeded overnight onto poly-D-lysine coated glass coverslips in 24-well plates and then stimulated with 10 ng/mL recombinant mouse IL-1α (R&D), 10 ng/mL recombinant mouse TGF-β (R&D), or media alone for 48 h. Coverslips were fixed in 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 15 min, blocked in 1% BSA for 30 min, and then stained overnight at 4 °C with anti α-SMA antibody (Invitrogen, clone IA4).

Techniques: Infection, Luciferase, Injection, Luminex

Journal: Scientific Reports

Article Title: T. gondii infection induces IL-1R dependent chronic cachexia and perivascular fibrosis in the liver and skeletal muscle

doi: 10.1038/s41598-020-72767-0

Figure Lengend Snippet: Cells expressing IL-1⍺ and IL-1R are observed in the fibrotic liver environment. ( a ) Cytokines in tissue lysates from mice at 9 wpi were measured by ELISA in liver. Data are presented as fold change relative to the mean of uninfected levels. N = 11–12 mice per group, pooled from three independent experiments. ( b ) IL-1α levels in the sera at 9 wpi measured by ELISA. N = 9–14 mice per group, pooled from four independent experiments. ( c ) Immunofluorescence labeling of nuclei (DAPI white), IL-1⍺ (green), CD45 (red), and collagen1⍺1 (blue) in the liver of UI or 9 wpi WT mice . Number of cells staining positive for CD45 and/or IL-1⍺, average 2–3 fields of view where immune infiltrate was present from 3 mice per condition are quantified on the right. Error bars are standard deviation. ( d ) Immunofluorescent labeling of nuclei (DAPI white), ⍺-smooth muscle actin (green), IL-1R (red), and collagen1⍺ (blue) in the liver of uninfected or 9 wpi WT. Inset, arrow head represents ⍺-smooth muscle actin/IL-1R co-staining cells (arrow heads). Number of cells staining positive for IL-1R and/or ⍺-SMA, average of 4–8 fields of view from 3 mice are quantified on the right. Error bars are standard deviation. ( c , d ) represent maximum intensity projections of 9–13 μm thick z-stacks. Scale bar represents 50 μm. Error bars are standard error of the mean except where noted otherwise. *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired Student’s t test.

Article Snippet: 1 × 10 4 cells were seeded overnight onto poly-D-lysine coated glass coverslips in 24-well plates and then stimulated with 10 ng/mL recombinant mouse IL-1α (R&D), 10 ng/mL recombinant mouse TGF-β (R&D), or media alone for 48 h. Coverslips were fixed in 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 15 min, blocked in 1% BSA for 30 min, and then stained overnight at 4 °C with anti α-SMA antibody (Invitrogen, clone IA4).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Labeling, Staining, Standard Deviation

Journal: Journal of Veterinary Science

Article Title: Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

doi: 10.4142/jvs.2010.11.3.213

Figure Lengend Snippet: Induction of dermatitis and interleukin (IL)-10 plasmid DNA injection schedule. Dorsal regions of mice were shaved on day 0. The hairless dorsal regions and glabrous ears of mice were sensitized with 200 µL of 1% (w/v) DNCB solution on day 4. Three days after sensitization, the dorsal skin and ears were challenged with 150 µL of 0.2% (w/v) DNCB solution every 3 days. Following in vivo delivery of IL-10 plasmid DNA, mice were intradermally injected with 100 µg of IL-10 plasmid DNA in 100 µL of phosphate-buffered saline (PBS) or PBS alone (control) on days 1 and 8 of experiment.

Article Snippet: Forty-eight hours after passage, the cell culture fluids were collected and analyzed for expression of mouse IL-10 protein by the mouse IL-10 Quantikine ELISA Kit (R&D Systems, USA) according to the manufacturer's instructions.

Techniques: Plasmid Preparation, Injection, In Vivo, Saline

Journal: Journal of Veterinary Science

Article Title: Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

doi: 10.4142/jvs.2010.11.3.213

Figure Lengend Snippet: Clinical skin severity score in the control mice increased gradually with the number of cutaneous applications of DNCB and reached a peak at the end of experiment. A significant improvement in mice injected with IL-10 plasmid DNA had occurred at day 13 ( * p <0.05) and by day 16 was even more pronounced ( ** p <0.01) compared with the control mice (mean ± SD; n = 5).

Article Snippet: Forty-eight hours after passage, the cell culture fluids were collected and analyzed for expression of mouse IL-10 protein by the mouse IL-10 Quantikine ELISA Kit (R&D Systems, USA) according to the manufacturer's instructions.

Techniques: Injection, Plasmid Preparation

Journal: Journal of Veterinary Science

Article Title: Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

doi: 10.4142/jvs.2010.11.3.213

Figure Lengend Snippet: The skin features demonstrated visually a marked reduction in severity of the dermatitis and more rapid hair re-growth with IL-10 plasmid DNA injection. (A) Dermatitis induced and PBS injected mice, (B) IL-10 plasmid DNA injected mice after dermatitis induction.

Article Snippet: Forty-eight hours after passage, the cell culture fluids were collected and analyzed for expression of mouse IL-10 protein by the mouse IL-10 Quantikine ELISA Kit (R&D Systems, USA) according to the manufacturer's instructions.

Techniques: Plasmid Preparation, Injection

Journal: Journal of Veterinary Science

Article Title: Inhibitory effects of interleukin-10 plasmid DNA on the development of atopic dermatitis-like skin lesions in NC/Nga mice

doi: 10.4142/jvs.2010.11.3.213

Figure Lengend Snippet: The inhibitory effects of interleukin-10 plasmid DNA. The concentration of serum IL-10 in mice injected with IL-10 plasmid DNA was significantly higher than in control mice. * p < 0.05 vs . control, ** p < 0.01 vs . control.

Article Snippet: Forty-eight hours after passage, the cell culture fluids were collected and analyzed for expression of mouse IL-10 protein by the mouse IL-10 Quantikine ELISA Kit (R&D Systems, USA) according to the manufacturer's instructions.

Techniques: Plasmid Preparation, Concentration Assay, Injection

Journal: Nutrients

Article Title: Protective Effect of the Total Saponins from Rosa laevigata Michx Fruit against Carbon Tetrachloride-Induced Liver Fibrosis in Rats

doi: 10.3390/nu7064829

Figure Lengend Snippet: Effects of Rosa laevigata Michx fruit (RLTS) on the expressions of TLR4, MyD88, NF-κB, iNOS, COX-2, IL-10, TNF-α, IL-1β and IL-6 ( A–L ). ( II ) Normal control; ( III ) model; ( IV ) RLTS (70 mg/kg) + CCl 4 ; ( V ) RLTS (140 mg/kg) + CCl 4 ; ( VI ) RLTS (210 mg/kg) + CCl 4 . Values are expressed as mean ± SD ( n = 4). * p < 0.05, ** p < 0.01 vs. model group.

Article Snippet: IL-10 , Interleukin-10 , rabbit , 1:400 , Boster Biological Technology, Wuhan, China.

Techniques: