Journal: Journal for Immunotherapy of Cancer

Article Title: IL-6 signaling in macrophages is required for immunotherapy-driven regression of tumors

doi: 10.1136/jitc-2021-002460

Figure Lengend Snippet: IL-6 blockade impairs the therapeutic efficacy of a therapeutic cancer vaccine. (A) The serum level of IL-6 (pg/mL) in TC-1.contol and TC-1 IL-6 tumor-bearing mice at different time points after tumor challenge. (B) The serum level of IL-6 (pg/mL) in TC-1.contol and TC-1 IL-6 tumor-bearig mice with and without IL-6R blockade or isotype control at day 16 post-tumor challenge. Tumor outgrowth graphs of the untreated or SLP vaccinated TC-1.control (C) and TC-1.IL-6 (D) tumor-bearing mice treated with or without IL-6 or IL-6R antibodies as described in the material and methods. The number shown above the x-axis is the number of alive mice from the total. (E) The average tumor outgrowth (upper panel) and the mean response rate (%) according to RECIST criteria (upper panel) of the major groups shown in (D). Lower panel: NR, no response; PR, partial response; CR, complete response. Graphs indicate mean values with SEM (F) survival graph of the mice shown in (D). Data in (A, B) is representative of three independent experiments, yielding similar results. Data shown in C–F are pooled from four independent experiments with similar results. Significance was determined by Mann-Whitney U test in B and student unpaired t-test for difference in endpoint tumor size in the vaccinated groups and χ 2 for trend for response rate in (E). Significance was determined by a log-rank (Mantel–Cox) test in (F). *P<0.05; **p<0.01; ***p<0.001. IL-6, interleukin 6; SLP, synthetic long peptide.

Article Snippet: Mouse IL-6 DuoSet ELISA and Mouse IL-6R alpha DuoSet ELISA (R&DSystems) and were used to measure the amount of IL-6 and IL-6R.

Techniques: MANN-WHITNEY

Journal: Journal for Immunotherapy of Cancer

Article Title: IL-6 signaling in macrophages is required for immunotherapy-driven regression of tumors

doi: 10.1136/jitc-2021-002460

Figure Lengend Snippet: IL-6 axis blockade partially impairs the tumor-specific T cell response. The percentage of intratumoral CD45 + leukocytes within live cells (A), the mean percentage of CD8 + , CD4 + (without Tregs), Tregs and CD11b + cells within CD45 + cells (B) and the percentage of E7 HPV Tm + cells within CD45 + cells (C) in untreated and SLP vaccinated TC-1.control and TC-1.IL-6 tumor-bearing mice. (D) The percentage of E7 HPV Tm + cells within CD8 + T cells in blood on day 16–20 and 30 post-tumor challenge in SLP vaccinated TC-1.control and TC-1.IL-6 tumor-bearing mice with and without IL-6R blockade. Each dot represents data from an individual mouse. Graphs indicate mean values with SEM data are pooled from two independent experiments with similar results. Significance between the vaccinated groups in each tumor model was determined by unpaired Student’s t-test. *P<0.05; **p<0.01. IL-6, interleukin 6; ns, not significant; SLP, synthetic long peptide.

Article Snippet: Mouse IL-6 DuoSet ELISA and Mouse IL-6R alpha DuoSet ELISA (R&DSystems) and were used to measure the amount of IL-6 and IL-6R.

Techniques:

Journal: Journal for Immunotherapy of Cancer

Article Title: IL-6 signaling in macrophages is required for immunotherapy-driven regression of tumors

doi: 10.1136/jitc-2021-002460

Figure Lengend Snippet: The composition of intratumoral myeloid cells is grossly unchanged with IL-6 axis blockade. The percentage of intratumoral CD11b + myeloid cells (A), cDC2, moDCs, immature monocytes and monocytes (B), macrophages (C) and pDCs and cDC1 (E) in SLP vaccinated or untreated TC-1control and TC-1.IL-6 tumor-bearing mice with and without IL-6 blockade. Graphs indicate mean values with SEM each dot represents data from an individual mouse. Data are representative of one experiment. (D) The expression of CD86, CD40 and MHC class II on macrophages. Significance in differences in the percentage of the indicated vaccine-associated cell infiltration due to IL-6R blockade were determined by Student’s t-test within each tumor model (A–E). *P<0.05; **p<0.01; ***p<0.001. IL-6, interleukin 6; mo DCs, monocyte-derived dendritic cells; ns, not significant; pDC, plasmacytoid DCs.

Article Snippet: Mouse IL-6 DuoSet ELISA and Mouse IL-6R alpha DuoSet ELISA (R&DSystems) and were used to measure the amount of IL-6 and IL-6R.

Techniques: Expressing, Derivative Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: IL-6 signaling in macrophages is required for immunotherapy-driven regression of tumors

doi: 10.1136/jitc-2021-002460

Figure Lengend Snippet: IL-6 blockade decreases the levels of SOCS3 and increases SIRPα levels. (A) The percentage of intratumoral SOCS3 + CD11b + (myeloid cells) and SOCS3 + F4/80 + (macrophages) cells and mean fluorescent intensity (MFI) of SOCS3 on macrophages. (B) MFI of pSTAT3 and pSTAT1 on total CD11b + myeloid cells and F4/80 + CD11b + macrophages. (C) MFI of SIRPα on F4/80 + CD11b + macrophages and CD11b - CD103 + (cDC1) cells. Scatter plots indicate mean values with SEM each dot represents data from an individual mouse. Violin plots show the full distribution of the data, each dot representing an individual mouse. Data are representative of one experiment. Significance (A–C) in differences in the indicated vaccine-associated percentage of indicated cell infiltration or mean fluorescence intensity of indicated markers on cells due to IL-6R blockade was determined by Student’s t-test within each tumor model. Significance between groups in D is determined by one-way ANOVA. *P<0.05; **p<0.01; ****p<0.0001. ANOVA, analysis of variance; SIRPα, signal-regulatory protein alpha; SOCS3, suppressor of cytokine signaling 3.

Article Snippet: Mouse IL-6 DuoSet ELISA and Mouse IL-6R alpha DuoSet ELISA (R&DSystems) and were used to measure the amount of IL-6 and IL-6R.

Techniques: Fluorescence

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 1. sIL-6R/p28 FP exerts stronger antiviral activity than sIL-6R alone. (A) The map of the sIL-6R/p28 FP con- struction is shown (upper panel). HEK293T cells were trans- fected with PKH3-33HA or PKH3-33HA-sIL-6R/p28 FP plasmids for 48 h. The expression of sIL-6R/p28 FP was de- tected by Western blot using the HA tag-specific Ab (lower panel). (B) A549 cells were transfected with control vector, sIL- 6R/p28 FP, or sIL-6R, p28, or both for 24 h and were then infected with IAV (MOI = 1) for another 24 h. Cells were harvested for total RNA isolation. Relative levels of NP-specific mRNA, cRNA, and vRNA were measured by qRT-PCR. **p , 0.01. (C) RD cells were transfected with control vector, sIL-6R/ p28 FP, or sIL-6R, p28, or both for 24 h. Twelve hours after EV71 (MOI = 1) infection, VP1 protein levels were determined by Western blot. Numbers below the blots are the quantified OD; control blots were set as 10. (D) Huh7 cells were cotransfected with pHBV1.3 (ayw) and control vector, sIL-6R/ p28 FP, or sIL-6R, p28, or both. Forty-eight hours after trans- fection, HBV capsid-associated DNA levels were assessed by qRT-PCR. **p , 0.01. (E) A549 cells were transfected with control vector, sIL-6R/p28 FP, or sIL-6R/p28 FP and sgp130 for 24 h and were then infected with IAV (MOI = 1) for another 24 h. Cells were harvested for total RNA isolation. Relative levels of NP-specific mRNA, cRNA, and vRNA were measured by qRT-PCR. All graphs depict the means and SDs for three experiments.

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Activity Assay, Expressing, Western Blot, Transfection, Control, Plasmid Preparation, Infection, Isolation, Quantitative RT-PCR

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 2. sIL-6R/p28 FP potentiates virus-triggered IFN and ISG induction. (A) A549 cells were cotransfected with IFN-b, IFN-l1, NF-kB, and ISRE (0.2 mg) luciferase reporter con- structs and pRL-TK (0.2 mg) together with sIL-6R/p28 FP or control vector (0.6 mg) expression plasmids. Twenty-four hours after transfection, cells were infected with Sendai virus (MOI = 1) for 12 h before reporter assays were performed. pRL-TK was used as an internal control. *p , 0.05, **p , 0.01. (B and C) A549 cells were transfected with vector or sIL-6R/p28 FP ex- pression plasmids for 24 h and were then infected with IAV (MOI = 1) for 24 h. Levels of IFN-a, IFN-b, IFN-l1 (B), OAS1, PKR, and MxA (C, left) were measured by qRT-PCR. OAS1, PKR, and MxA proteins were assessed by Western blot, re- spectively (C, right). *p , 0.05. Numbers below the blots are the quantified OD; control blots were set as 10. (D) A549 cells were transfected with control vector, sIL-6R/p28 FP, or sIL-6R/ p28 FP and sgp130 for 24 h and were then infected with IAV (MOI = 1) for 24 h. Levels of IFN-a, IFN-b, and IFN-l1 were measured by qRT-PCR. (E) A549 cells were transfected with vector or sIL-6R/p28 FP expression plasmids for 24 h and were then infected with IAV (MOI = 1) for 24 h. IL-1b, IL-8, TNF-a, and IL-6 mRNA levels were measured by qRT-PCR. All graphs represent the means and SDs for three experiments. *p , 0.05.

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Virus, Luciferase, Control, Plasmid Preparation, Expressing, Transfection, Infection, Quantitative RT-PCR, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 3. sIL-6R is required for the virus-triggered immune response. (A) A549 cells were used for constructing IL-6R-KO cell lines using the CRISPR-Cas9 system. A549 wild-type cells and two chosen monoclonal KO cells were ex- amined by Western blot using an IL-6R–specific Ab. (B–D) The same numbers of Scr.-WT and IL-6R-KO cells were infected with IAV (MOI = 1) for 24 h, and cells were harvested for total RNA isolation. Relative levels of NP-specific mRNA, cRNA, and vRNA were measured by qRT-PCR (B). Levels of IFN-a, IFN-b, IFN-l1 (C), OAS1, PKR, and MxA (D) were measured by qRT-PCR. *p , 0.05, **p , 0.01. (E–G) IL-6R-KO cells were transfected with vector or sIL-6R expression plasmids for 24 h and then infected with IAV (MOI = 1) for 24 h. Cells were harvested for total RNA isolation. Relative levels of NP-specific mRNA, cRNA, and vRNA were measured by qRT-PCR. (E) Equal numbers of Scr.-WT and IL-6R-KO A549 cells were infected with IAV (MOI = 1) for 24 h. IL-1b, IL-8, TNF-a, and IL-6 mRNA levels were measured by qRT-PCR. *p , 0.05, **p , 0.01. (F) Levels of IFN-a, IFN-b, IFN-l1 (G), OAS1, PKR, and MxA (H) were measured by qRT-PCR. *p , 0.05, **p , 0.01. (I) IL-6R-KO A549 cells were transfected with vector or sIL-6R expression plasmids for 24 h and were then infected with IAV (MOI = 1) for 24 h. IL-1b, IL-8, TNF-a, and IL-6 mRNA levels were measured by qRT-PCR. All graphs represent the means and SDs for three experiments. *p , 0.05, **p , 0.01.

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Virus, CRISPR, Western Blot, Infection, Isolation, Quantitative RT-PCR, Transfection, Plasmid Preparation, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 4. sIL-6R/p28 FP inter- acts with TRAF6 and MAVS. A549 WT cells or IL-6R-KO cells were transfected with IFN-b (A), ISRE (B), or NF-kB (C) luciferase reporter constructs (0.2 mg) along with pRL- TK (0.2 mg), and the indicated ex- pression plasmids (0.6 mg) for 24 h before reporter assays were performed. pRL-TK was used as an internal con- trol. *p , 0.05. HEK293T cells were cotransfected with Flag-sIL-6R/p28 FP and HA-TRAF6 expression plas- mids (D), HA-sIL-6R/p28 FP and Flag-MAVS expression plasmids (E), Flag-sIL-6R and HA-TRAF6 expres- sion plasmids (F), and Flag-p28 and HA-TRAF6 expression plasmids (G). HA-sIL-6R and Flag-MAVS expres- sion plasmids (H), or HA-p28 and Flag-MAVS expression plasmids (I) for 48 h. Coimmunoprecipitation and immunoblot analyses were performed with the indicated Abs. All experi- ments were repeated at least three times with consistent results.

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Transfection, Luciferase, Construct, Expressing, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 5. Intracellular sIL-6R and p28 colocalize with TRAF6 and MAVS. (A) Colocalization of p28 and MAVS was detected by confocal microscopy. A549 cells were left untreated or infected with IAV (MOI = 1) for 12 h, and then p28/MAVS and nuclei were stained with Dylight 488/Cy3-conjugated secondary Abs and DAPI, respectively. (B) Colocalization of sIL-6R and TRAF6 was detected by confocal microscopy. A549 cells were transfected with HA-sIL-6R for 24 h, and then cells were left untreated or infected with IAV (MOI = 1) for 12 h. Then TRAF6/sIL-6R and nuclei were stained with FITC/Cy3- conjugated secondary Abs and DAPI, respectively. Scale bars, 10 mm. All experiments were repeated at least three times with consistent results.

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Confocal Microscopy, Infection, Staining, Transfection

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 6. The sIL-6R/p28 FP and MAVS/TRAF6 complex affects the expression of IFN-l1. (A) Stable IFNAR1-KD and IFN-lR1-KD were collected to assess IFNAR1 and IFN-lR1 protein levels by Western blot, respectively. Scr.-KD A549 cells were used as a control. (B) IFNAR1-KD, IFN-lR1-KD, and Scr.-KD A549 cells were transfected with sIL-6R/p28 FP or control vector expression plasmids for 24 h and then infected with IAV (MOI = 1) for another 24 h. Cells were harvested for total RNA isolation. Relative levels of NP-specific mRNA, cRNA, and vRNA were measured by qRT-PCR. *p , 0.05, **p , 0.01. (C) A549 cells were transfected with small inter- fering MAVS, small interfering TRAF6, or small interfering NC for 48 h. Cells were collected to assess MAVS and TRAF6 protein levels by Western blot (left). IFNAR1-KD A549 cells were cotransfected with vector or sIL-6R/p28 FP (1 mg) and small interfering MAVS, small interfering TRAF6, or small interfering NC (50 nM). Twenty-four hours after transfection, cells were infected with IAV (MOI = 1) for another 24 h before total RNA isolation. Relative levels of NP-specific mRNA, cRNA, and vRNA were measured by qRT-PCR. *p , 0.05 (right). (D) Schematic representation of the IFN-l1 promoter is shown. ChIP primer sets are also shown (upper panel). IL-6R- KO and Scr.-WT A549 cells were infected with IAV (MOI = 1) for 12 h. ChIP assays for binding of p50 and p65 to the NF-kB site (for ChIP primer location F1/R1) or IRF3 and IRF7 to ISRE site (for ChIP primer location F2/R2) were performed. Numbers below the blots are the quantified OD; control blots were set as 10. All experiments were repeated at least three times with similar results. Graphs represent means 6 SD, n = 3. *p , 0.05 (lower panel).

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Expressing, Western Blot, Control, Transfection, Plasmid Preparation, Infection, Isolation, Quantitative RT-PCR, Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 8. sIL-6R/p28 FP regulates IFN-l1 gene expression through p38 MAPK. (A) A549 cells were transfected with vector or sIL-6R/p28 FP expression plasmids for 24 h and were then infected with IAV (MOI = 1) for another 12 h. ChIP assays for binding of c-Fos and ATF1 were performed. *p , 0.05. (B) A549 cells were treated with SB 203580 (10 mM) or DMSO for 1 h and were then infected with IAV (MOI = 1) for another 12 h. ChIP assays for binding of c-Fos and ATF1 were per- formed. *p , 0.05. (C) A549 cells were transfected with vector or sIL-6R/p28 FP expression plasmids for 24 h before treatment with SB 203580 (10 mM) or DMSO for 1 h. Then cells were infected with IAV (MOI = 1) for another 12 h. Levels of IFN-l1 were measured by qRT-PCR. *p , 0.05. (D) A549 cells were transfected with vector or sIL-6R/p28 FP expression plasmids for 24 h and then infected with IAV (MOI = 1) for the indicated times. Cell lysates were examined by Western blot with p38 phosphospecific and total Abs. (E) A549 cells were transfected with vector or sIL-6R/p28 FP expression plasmids for 24 h and then infected with IAV (MOI = 1) for another 12 h (left). A549 cells were left untreated or treated with SB 203580 (10 mM) or DMSO for 1 h before infection with IAV (MOI = 1) for another 12 h (right). Cell lysates were examined by Western blot with c-Fos–specific Ab. (F) A549 cells were transfected with vector or sIL-6R/p28 FP expression plasmids for 24 h and were then infected with IAV (MOI = 1) for 1 h (left). Cells were left untreated or were treated with SB 203580 (10 mM) or DMSO for 1 h and then were infected with IAV (MOI = 1) for another 1 h (right). Cell lysates were examined by Western blot with ATF1 phosphospecific and total Abs. Numbers below the blots are the quantified OD; control blots were set as 10. All exper- iments were repeated at least three times with similar results.

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Gene Expression, Transfection, Plasmid Preparation, Expressing, Infection, Binding Assay, Quantitative RT-PCR, Western Blot, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Soluble IL-6 Receptor and IL-27 Subunit p28 Protein Complex Mediate the Antiviral Response through the Type III IFN Pathway.

doi: 10.4049/jimmunol.1600627

Figure Lengend Snippet: FIGURE 9. Schematic of the proposed model for sIL-6R/p28 FP-mediated type I and type III IFN expression. Solid arrows represent signaling pathways identified in this study or in a previous study. Broken arrows indicate potential signaling pathways. Viruses (IAV, EV71, and HBV) induce sIL-6R and p28 expression. The sIL-6R/p28 FP associates with the MAVS/TRAF6 complex, leading to activation of type I IFN. In contrast, the sIL-6R/p28 FP and MAVS/TRAF6 complex induces IFN-l1 expression through regulation of the binding of IRF3, p50, and p65 to the IFN-l1 promoter. At the same time, sIL-6R/p28 FP induces the p38 MAPK signaling pathway, leading to the activation of c-Fos and ATF1, which bind to the IFN-l1 promoter and activate IFN-l1 expression.

Article Snippet: Abs against IL-6R, p28, TRAF6,MAVS, 29-59-oligoadenylate synthetase 1 (OAS1), PKR, IFNAR1, IFN-lRl, p65, p50, IRF3, IRF7, ATF1, and c-Fos (Santa Cruz Biotechnology); myxovirus resistance A (MxA) and VP1 (ProteinTech Group); p-ATF1, p38, and p-p38 (Cell Signaling Technology); hemagglutinin (HA) and Flag (Medical and Biological Laboratories, MBL); GAPDH, b-actin, and b-tubulin (Invitrogen) were purchased from the indicated manufacturers.

Techniques: Expressing, Protein-Protein interactions, Activation Assay, Binding Assay

Journal: Journal of Clinical Medicine

Article Title: Characteristics of Interleukin-6 Signaling in Elective Cardiac Surgery—A Prospective Cohort Study

doi: 10.3390/jcm11030590

Figure Lengend Snippet: Exaggeration of IL-6, sIL-6R and sgp130. The figure displays individual time course ( left ) and corresponding boxplots ( right ) of perioperative levels of IL-6, sIL-6R and sgp130 levels. The lines on the left are displayed on a a logarithmic scale. Asterisks (*) mark significant differences compared to baseline at p less than 0.05. In the boxplots, the lower boundary of the box indicates the 25th percentile, a black line within the box marks the median, and the upper boundary of the box indicates the 75th percentile. Whiskers above and below the box indicate the 10th and 90th percentiles. Points (°) above and below the whiskers indicate outliers outside the 10th and 90th percentiles (IL-6 outliers above 1200 pg/mL are not shown). Red, blue and green lines indicate IL-6, sIL-6R and sgp130 levels, respectively. Abbreviations: CPB, cardiopulmonary bypass; POD, postoperative day; sgp130, soluble glycoprotein 130; sIL-6R, soluble interleukin-6 receptor.

Article Snippet: Quantitative measurements of IL-6, sIL-6R, and sgp130 concentrations were conducted with 96-well plate ELISAs (Human IL-6 Quantikine ELISA Kit, Human IL-6R alpha Quantikine ELISA Kit and Human soluble gp130 Quantikine ELISA Kit, R&D Systems ® Inc., Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Characteristics of Interleukin-6 Signaling in Elective Cardiac Surgery—A Prospective Cohort Study

doi: 10.3390/jcm11030590

Figure Lengend Snippet: sIL-6/IL-6 and sIL-6R/sgp130 ratio. The lines show the perioperative levels of ( a ) sIL-6/IL-6 and ( b ) sIL-6R/sgp130 ratio at different time points. Red lines indicate patients with continuously lowest levels, blue lines indicate patients with continuously highest levels and grey lines indicate patients with intermediate levels of perioperative sIL6-R/IL-6 ratio measured between timepoint 3 and 9. Abbreviations: CPB, cardiopulmonary bypass; POD, postoperative day; sgp130, soluble glycoprotein 130; sIL-6R, soluble interleukin-6 receptor.

Article Snippet: Quantitative measurements of IL-6, sIL-6R, and sgp130 concentrations were conducted with 96-well plate ELISAs (Human IL-6 Quantikine ELISA Kit, Human IL-6R alpha Quantikine ELISA Kit and Human soluble gp130 Quantikine ELISA Kit, R&D Systems ® Inc., Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: