Journal: Frontiers in Immunology

Article Title: The TNFR1 Antagonist Atrosimab Is Therapeutic in Mouse Models of Acute and Chronic Inflammation

doi: 10.3389/fimmu.2021.705485

Figure Lengend Snippet: Atrosimab blocks acute TNF-induced inflammation in vivo. (A–C) C57BL/6 huTNFR1-k/i mice received i.v. injections of TNF (0.3 mg/kg), Atrosimab or FcΔab (30 mg/kg). (A) Body weight was determined and (B, C) blood samples were collected before treatment (0 h) and 1 h, 6 h, 24 h and 72 h after the injections. (B) IL-6 and (C) CRP levels in the serum were determined by ELISA. (D–F) C57BL/6 huTNFR1-k/i mice received i.v. injections of PBS, Atrosimab (30 mg/kg body weight) or FcΔab (30 mg/kg). TNF (0.3 mg/kg) was injected 30 minutes thereafter to induce acute inflammatory responses. (D) Body weight was determined and (E, F) blood samples were collected before treatment (0 h) and 1 h, 6 h, 24 h and 72 h after TNF injection. (E) IL-6 and (F) CRP levels in the serum were determined by ELISA. Individual values were excluded using the ROUT method of outlier identification or because they were below the detection limit (Q = 1%) or because they were below the detection limit of the ELISA. Mean ± SD, n=6. (A, C) : **p < 0.01; (D) : *p < 0.05 – TNF + PBS vs TNF + Atrosimab, # p < 0.05 – TNF + PBS vs TNF + FcΔab; (E, F) : ns, not significant, **p < 0.01.

Article Snippet: At the end of the study, serum samples were analyzed for IL-6 (IL-6 Duo-Set, R&D, DY406) according to the manufacturer’s instructions.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Injection

Journal: Frontiers in Immunology

Article Title: Short-Term High-Fat Diet Consumption Reduces Hypothalamic Expression of the Nicotinic Acetylcholine Receptor α7 Subunit (α7nAChR) and Affects the Anti-inflammatory Response in a Mouse Model of Sepsis

doi: 10.3389/fimmu.2019.00565

Figure Lengend Snippet: Cytokine levels in the hypothalamus and peripheral tissues following i.p. LPS challenge and i.p. PNU-282987 stimulation. (A) mRNA expression levels of TNFα (SC, n = 5 and HFD, n = 5); (B) mRNA expression levels of IL-1β (SC, n = 5 and HFD, n = 5); (C) mRNA expression levels of IL-6 (SC, n = 5 and HFD, n = 5); in the hypothalamus; (D) mRNA levels of TNFα (SC, n = 5 and HFD, n = 5); (E) mRNA expression levels of IL-1β (SC, n = 5 and HFD, n = 5); (F) mRNA expression levels of IL-6 (SC, n = 5 and HFD, n = 5) in the spleen; (G) mRNA expression levels of TNFα (SC, n = 5 and HFD, n = 5); (H) mRNA expression levels of IL-1β (SC, n = 5 and HFD, n = 5); (I) mRNA expression levels of IL-6 (SC, n = 5 and HFD, n = 5) in the liver; of mice fed SC or a HFD for 3 days and injected intraperitoneally with a high dose of LPS (12 mg/kg) followed by stimulation with PNU-282987 (3 mg/kg). The bars represent the mean ± S.E.M. * Means significantly different as shown by one-way ANOVA ( * p < 0.05; ** p < 0.01; # p < 0.05; § p < 0.05). # and § to compare SC and HFD.

Article Snippet: The concentrations of cytokines in hypothalamus and serum (TNF-α and IL-1β) in the supernatants were assessed by ELISA using the Duo Set kit (R&D System) and IL6 using IL-6 mouse ELISA kit (Thermofischer Scientific).

Techniques: Expressing, Injection

Journal: Frontiers in Immunology

Article Title: Short-Term High-Fat Diet Consumption Reduces Hypothalamic Expression of the Nicotinic Acetylcholine Receptor α7 Subunit (α7nAChR) and Affects the Anti-inflammatory Response in a Mouse Model of Sepsis

doi: 10.3389/fimmu.2019.00565

Figure Lengend Snippet: Chemokine and cytokine levels in the hypothalamus following i.p. LPS challenge and i.c.v. PNU-282987 stimulation. (A) mRNA expression levels of CX3CL1 (SC, n = 6 and HFD, n = 6); (B) mRNA expression levels of CCL2 (SC, n = 6 and HFD, n = 6); (C) mRNA expression levels of TNFα (SC, n = 5 and HFD, n = 5); (D) mRNA expression levels of IL-1β (SC, n = 6 and HFD, n = 6); and (E) mRNA expression levels of IL-6 (SC, n = 6 and HFD, n = 6) in the hypothalamus of mice fed SC or a HFD for 3 days and injected intraperitoneally with a high dose of LPS (12 mg/kg) followed by stimulation intracerebroventricularly with PNU-282987 (10 pmoles/mouse). The bars represent the mean ± S.E.M. *Means significantly different as shown by one-way ANOVA (* p < 0.05; ** p < 0.01; # p < 0.05; ## p < 0.001; ### p < 0,01; § p < 0.05). #, ##, ### and §to compare SC and HFD.

Article Snippet: The concentrations of cytokines in hypothalamus and serum (TNF-α and IL-1β) in the supernatants were assessed by ELISA using the Duo Set kit (R&D System) and IL6 using IL-6 mouse ELISA kit (Thermofischer Scientific).

Techniques: Expressing, Injection

Journal: Frontiers in Immunology

Article Title: Short-Term High-Fat Diet Consumption Reduces Hypothalamic Expression of the Nicotinic Acetylcholine Receptor α7 Subunit (α7nAChR) and Affects the Anti-inflammatory Response in a Mouse Model of Sepsis

doi: 10.3389/fimmu.2019.00565

Figure Lengend Snippet: Chemokine and cytokine levels in the hypothalamus following sepsis induction by CLP. (A) mRNA expression levels of CCL2 (SC, n = 5 and HFD, n = 5); (B) mRNA expression levels of CX3CL1 (SC, n = 6 and HFD, n = 6); (C) mRNA expression levels of IL-1β (SC, n = 6 and HFD, n = 6); (D) mRNA expression levels of TNFα (SC, n = 6 and HFD, n = 6); (E) mRNA expression levels of IL-6 (SC, n = 6 and HFD, n = 6); (F) mRNA expression levels of Chil3 (SC, n = 6 and HFD, n = 6); and (G) mRNA expression levels of IL-10 (SC, n = 6 and HFD, n = 6) in the hypothalamus of mice fed SC or a HFD for 3 days followed by induction of sepsis by cecal ligation and puncture surgery. The bars represent the mean ± SEM. * Means significantly different as shown by one-way ANOVA ( * p < 0.05; ** p < 0.01; *** p < 0.0001).

Article Snippet: The concentrations of cytokines in hypothalamus and serum (TNF-α and IL-1β) in the supernatants were assessed by ELISA using the Duo Set kit (R&D System) and IL6 using IL-6 mouse ELISA kit (Thermofischer Scientific).

Techniques: Expressing, Ligation

Journal: bioRxiv

Article Title: Pharmacological CDK4/6 inhibition unravels a p53-induced secretory phenotype in senescent cells

doi: 10.1101/2020.06.05.135715

Figure Lengend Snippet: p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). N=6 mice/group. 14 dpt, bioluminescence was visualized and quantified by the IVIS spectrum in vivo imaging system, as shown by representative bioluminescence images ( A ) and quantification ( B ). ( C ) RNA isolated from treated kidneys and mRNA encoding p16 quantified by qRT-PCR. Representative images ( D ) to visualize SA-β-gal activities in vehicle-, doxorubicin-or abemaciclib-treated mouse kidney sections at 15 dpt (arrows indicated positive area; scale bar, 1 mm; N=3) and quantified ( E ). ( F ) 15 dpt, plasma was collected and expression levels of CXCL1 measured by ELISA. ( G ) Protein lysate obtained from drug treated kidneys to quantify IL6 by ELISA. ( H and I ) RNA isolated from kidneys and mRNA encoding indicated genes quantified by qRT-PCR. ( J ) Relative weight changes were calculated at 7 dpt and 14 dpt. Red blood cells ( K ) and white blood cells ( L ) were counted at 15 dpt. Percentage of T cells, B cells, granulocytes and macrophages were determined by flow cytometry analysis ( M ). Physical performance was measured by rotarods assay at 15 dpt ( N ), grip strength meter at 7 dpt and 14 dpt ( O ), and hanging tests were performed at 7 dpt and 14 dpt and normalized to weights ( P ). One-way ANOVA, data are means ±SD ( B, C, E, F, G, K, L and N ). Two-way ANOVA, data are means ±SD ( H, I, J, M, O and P ). *p<0.05, **p<0.01, ***p<0.001, N.S.=not significant. dpt, days post treatment.

Article Snippet: 20 μg total protein was collected from each treated kidney and IL6 protein level was determined by the mouse IL6 duo-set (R&D Systems) ELISA.

Techniques: In Vivo Imaging, Isolation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Breast Cancer Research : BCR

Article Title: Attenuated transforming growth factor beta signaling promotes metastasis in a model of HER2 mammary carcinogenesis

doi: 10.1186/s13058-014-0425-7

Figure Lengend Snippet: Cytokine and chemokine profile of Erbb2+ mammary carcinoma cells with intact and modified transforming growth factor β (TGFβ) signaling. (A) Thymidine incorporation assay. Epithelial cells were treated with 0.5, 1 and 5 ng/mL TGFβ and analyzed for changes in cell proliferation by 3 (H) thymidine incorporation. Statistical significance was determined by P -values <0.05; *202Mul treated versus 202Mul untreated, **202Mul/DN treated versus 202Mul/DN untreated, ***202Mul/DN treated versus 202Mul treated. (B) Representative mouse cytokine array using tumor explant supernatants from 202Mul and 202Mul/DN cell lines (left) quantification of the mouse cytokine array data for IL-6, monocyte chemotactic protein 1 (MCP1) and tissue inhibitor of metalloproteinase 1 (TIMP1) (right). (C) ELISA of conditional medium from 202Mul and 202Mul/DN cell lines untreated and treated for 24 hours with TGFβ 1 ng/mL. All changes in cytokine and chemokine secretion were significant ( P <0.05) in 202Mul versus 202Mul/DN cell lines and treated versus untreated with TGFβ. (D) ELISA of protein extracts from tumor tissue homogenates isolated from NK1Mul and NK1Mul/DN mice on 5 and 11 weeks after tumor palpation. Five mice per group were used excluding vascular endothelial growth factor (VEGF) measurement, where eight mice were used; * P <0.05. Data correspond to the mean ± standard error of the mean.

Article Snippet: Cytokine levels in conditional media and tissue lysates were measured using the mouse CXCL1, CXCL5, MCP-1, VEGF, and IL-6 ELISA Duo kits (R&D Systems, Minneapolis, MN, USA) following the manufacturer's protocol.

Techniques: Modification, Thymidine Incorporation Assay, Enzyme-linked Immunosorbent Assay, Isolation