Journal: Pain Physician

Article Title: Evaluation of the Effect of Duration on the Efficacy of Pulsed Radiofrequency in an Animal Model of Neuropathic Pain

doi: 10.36076/ppj.2018.2.191

Figure Lengend Snippet: Fig. 1. Immunopositive nerve fibers for TNF-α of CCI rats after scarification.

Article Snippet: Paraformaldehyde-fixed paraffin-embedded sciatic nerve sections (5 mm thickness) were incubated with antibodies to TNF-α (MP6-XT22, Novus Biologicals, Littleton, CO) and IL-6 (20F3, Thermo Fisher Scientific, Waltham, MA).

Techniques:

Journal: Experimental cell research

Article Title: Using phage-assisted continuous evolution (PACE) to evolve human PD1.

doi: 10.1016/j.yexcr.2020.112244

Figure Lengend Snippet: Fig. 3. Summarization of PD1 variants that bind PDL1. (A) Under high levels of mutagenesis (MP6) during the 288 h of PACE. The Red line indicates the phage titer of each time point. And the grey line indicates the flow velocity of system. (B) During 288 h of evolution, “lagoon” samples were sequenced using the Sanger method, and 11 different preponderant oligotypes were identified which contained 9 amino acid sites. (C) Among these 11 oligotypes, 9 amino acids exhibited diverse retention from 9 times of T98L to 3 times G18A. (D) The summarization of 11 oligotypes is shown on a time axis.

Article Snippet: SP098, pAB107a, MP6 plasmid, S2060 and S2208 were obtained from Addgene.

Techniques: Mutagenesis

Journal: Journal of Virology

Article Title: Distinct Roles of Vaccinia Virus NF-κB Inhibitor Proteins A52, B15, and K7 in the Immune Response

doi: 10.1128/JVI.00575-17

Figure Lengend Snippet: Deletion of the A52R, B15R, and K7R genes influences adaptive HIV-specific CD8 T cell immune responses. Shown are the vaccine-induced HIV-specific CD8 T cell response in mice (n = 4/group) immunized following a heterologous DNA prime/virus boost regimen (107 PFU of NYVAC-C or NYVAC-C deletion mutants). (A) Percentages of TNF-α+ CD107a+, IFN-γ+ TNF-α+, IFN-γ+ CD107a+ double-positive and IL-2 single-positive T cells. The total value (magnitude) is the sum of percentages of CD8 T cells that express IFN-γ and/or TNF-α and/or IL-2 and/or CD107a, measured by intracellular cytokine staining (ICS). Nonspecific responses of mice infected with control NYVAC-WT were subtracted from the total magnitude. (B to D) Magnitude of Pol-1 (B)-, Gag-Pool (C)-, or Pol-2 (D)-specific CD8 T cell responses. Graphs show the mean ± confidence interval (CI). (E) Functional profile of adaptive Gag-Pol-specific CD8 T cells. Combinations of responses (x axis) and percentages of functionally distinct cell subsets (y axis) are shown in the bar graph. Responses are grouped and color coded based on the number of functions. Pie chart colors indicate the percentage of cytokine-producing cells based on the number of functions (inside) and the different activation markers (outside). Data are representative of two independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: For intracellular cytokine staining (ICS), splenocytes were resuspended in RPMI 1640 with 10% FCS and 1 μg/ml Golgiplug (BD), monensin (eBioscience), and anti-CD107a (1D4B; BD), restimulated with peptides (6 h, 37°C, 5% CO 2 ), stained for surface markers with anti-CD3 (145-2C11), anti-CD4 (GK1.5), and anti-CD8 (53-6.7) (all from BD), fixed, permeabilized (Cytofix/Cytoperm kit; BD), and stained intracellularly with anti-IL-2 (JES6-5H4), anti-IFN-γ (XMG 1.2), and anti-TNF-α (MP6-X722) (all from BD).

Techniques: Staining, Infection, Functional Assay, Activation Assay

Journal: JCI Insight

Article Title: TNF and granulocyte macrophage-colony stimulating factor interdependence mediates inflammation via CCL17

doi: 10.1172/jci.insight.99249

Figure Lengend Snippet: (A and B) WT mice received an intra-articular (i.a.) injection of zymosan and anti-TNF mAb, anti–granulocyte macrophage-colony stimulating factor (anti–GM-CSF) mAb, IgG1 or IgG2a isotype control mAbs (150 μg i.p.), either (A) prophylactically (on days –2 and 0) (n = 4–5 mice/group) or (B) therapeutically (day 1) (n = 5–10 mice/group), and pain and arthritis (histology, day 7) were measured. Mice were also treated (B) therapeutically with anti-CCL17 mAb (150 μg i.p.) (day 1) (n = 5 mice/group). For histology images, original magnification ×60. Results are shown as mean ± SEM. P values were obtained using a 2-way ANOVA test for pain (weight distribution) readings, and a 1-way ANOVA test for histology. *P < 0.05, ***P < 0.001, ****P < 0.0001, anti-TNF versus IgG1 isotype; #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001, anti–GM-CSF versus IgG2a isotype; ζP < 0.05, ζζP < 0.01, anti-CCL17 versus IgG2a isotype.

Article Snippet: Cells were treated with either murine GM-CSF (20 ng/ml) or PBS, for the indicated time periods, in the presence or absence of anti-TNF mAb (MP6-XT22, 1 ng/ml) (Schering BioPharma) ( 76 ) or IgG1 isotype control mAb (1 ng/ml).

Techniques: Injection

Journal: JCI Insight

Article Title: TNF and granulocyte macrophage-colony stimulating factor interdependence mediates inflammation via CCL17

doi: 10.1172/jci.insight.99249

Figure Lengend Snippet: (A and B) Intraplantar (i.pl.) injection of granulocyte macrophage-colony stimulating factor (GM-CSF) (20 ng) or saline in (A) WT and Tnf–/– mice (n = 4–6 mice/group) and in (B) WT mice treated with anti-TNF or isotype control (2 μg/paw i.pl. at t = 0) (n = 12 mice/group). Pain was measured. (C) Methylated BSA (mBSA)/GM-CSF arthritis (intra-articular [i.a.] mBSA [day 0] and GM-CSF or saline s.c. [days 0–2]) was induced in WT and Tnf–/– mice. Pain and arthritis (histology, day 7) were measured (n = 4–6 mice/group). Original magnification, ×60. Results are shown as mean ± SEM. P values were obtained using a 2-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, WT saline versus WT GM-CSF; #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001, WT GM-CSF versus Tnf–/– GM-CSF.

Article Snippet: Cells were treated with either murine GM-CSF (20 ng/ml) or PBS, for the indicated time periods, in the presence or absence of anti-TNF mAb (MP6-XT22, 1 ng/ml) (Schering BioPharma) ( 76 ) or IgG1 isotype control mAb (1 ng/ml).

Techniques: Injection, Methylation

Journal: JCI Insight

Article Title: TNF and granulocyte macrophage-colony stimulating factor interdependence mediates inflammation via CCL17

doi: 10.1172/jci.insight.99249

Figure Lengend Snippet: (A and B) Methylated BSA (mBSA)/granulocyte macrophage-colony stimulating factor (GM-CSF) arthritis (intra-articular [i.a.] mBSA [day 0] and GM-CSF or saline s.c. [days 0–2]) was induced in WT mice treated with anti-TNF mAb or IgG1 isotype control (150 μg i.p.) (A) prophylactically (on days –2 and 0) (n = 8–12 mice/group) and (B) therapeutically (day 4) (n = 5 mice/group). Mice were also treated (B) therapeutically with anti-CCL17 mAb or IgG2a isotype control (150 μg i.p.) (day 4) (n = 4–5 mice/group). Pain (incapacitance meter) and arthritis (histology) were measured. Original magnification, ×125. Results are shown as mean ± SEM. P values were obtained using a 2-way ANOVA test for pain (weight distribution) readings and a Mann-Whitney U test or 1-way ANOVA for histology quantification. *P < 0.05, ***P < 0.001, ****P < 0.0001, saline versus GM-CSF + isotype; ####P < 0.0001, GM-CSF + isotype versus GM-CSF + anti-TNF; ζP < 0.05, ζζP < 0.01, GM-CSF + isotype versus GM-CSF + anti-CCL17.

Article Snippet: Cells were treated with either murine GM-CSF (20 ng/ml) or PBS, for the indicated time periods, in the presence or absence of anti-TNF mAb (MP6-XT22, 1 ng/ml) (Schering BioPharma) ( 76 ) or IgG1 isotype control mAb (1 ng/ml).

Techniques: Methylation, MANN-WHITNEY

Journal: JCI Insight

Article Title: TNF and granulocyte macrophage-colony stimulating factor interdependence mediates inflammation via CCL17

doi: 10.1172/jci.insight.99249

Figure Lengend Snippet: (A–D) Intraplantar (i.pl.) injection of TNF (20 ng) or saline in (A) WT mice treated with/without indomethacin (12.5 μg/paw i.pl. at 2 hours) (n = 10 mice/group), (B) WT mice treated with/without the COX-2 inhibitor, SC58125 (5 mg/kg i.p. at t = –30 minutes), or guanethidine sulfate (25 mg/kg s.c. at –60 minutes) for 4 hours (n = 5–8 mice/group), (C) WT and GMCSF–/– mice (n = 5–10 mice/group), and (D) WT mice treated with anti–GM-CSF mAb or isotype control (2 μg/paw i.pl. at t = 0) (n = 5 mice/group). Pain (incapacitance meter) was measured. Results are shown as mean ± SEM. P values were obtained using a 1-way (B) or 2-way (A, C, and D) ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, WT saline versus WT TNF or TNF + isotype; #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001, TNF versus TNF + indo or TNF + COX-2 inhibitor; WT TNF versus GMCSF–/– TNF; TNF + isotype vs. TNF + anti–GM-CSF. GM-CSF, granulocyte macrophage-colony stimulating factor.

Article Snippet: Cells were treated with either murine GM-CSF (20 ng/ml) or PBS, for the indicated time periods, in the presence or absence of anti-TNF mAb (MP6-XT22, 1 ng/ml) (Schering BioPharma) ( 76 ) or IgG1 isotype control mAb (1 ng/ml).

Techniques: Injection

Journal: JCI Insight

Article Title: TNF and granulocyte macrophage-colony stimulating factor interdependence mediates inflammation via CCL17

doi: 10.1172/jci.insight.99249

Figure Lengend Snippet: (A and B) Methylated BSA (mBSA)/TNF arthritis (intra-articular [i.a.] mBSA [day 0] and TNF or saline s.c. [days 0–2]) was induced in (A) WT mice treated prophylactically with anti–granulocyte macrophage-colony stimulating factor (anti–GM-CSF) mAb or isotype control (150 μg i.p. on days –2 and 0 i.p.) (n = 9–10 mice/group) and (B) WT mice treated therapeutically with anti–GM-CSF mAb or isotype control (150 μg i.p. on day 2 i.p.) (n = 4–5 mice/group). Pain and arthritis (histology) were measured. Original magnification, ×125. Results are shown as mean ± SEM. P values were obtained using a 2-way ANOVA test for pain (weight distribution) readings, and a 1-way ANOVA test for histology quantification. *P < 0.05, ****P < 0.0001, saline versus TNF + isotype; #P < 0.05, ##P < 0.01, ####P < 0.0001, TNF + isotype versus TNF + anti–GM-CSF.

Article Snippet: Cells were treated with either murine GM-CSF (20 ng/ml) or PBS, for the indicated time periods, in the presence or absence of anti-TNF mAb (MP6-XT22, 1 ng/ml) (Schering BioPharma) ( 76 ) or IgG1 isotype control mAb (1 ng/ml).

Techniques: Methylation

Journal: JCI Insight

Article Title: TNF and granulocyte macrophage-colony stimulating factor interdependence mediates inflammation via CCL17

doi: 10.1172/jci.insight.99249

Figure Lengend Snippet: (A and B) Methylated BSA (mBSA)/TNF arthritis (intra-articular [i.a.] mBSA [day 0] and TNF or saline s.c. [days 0–2]) was induced in (A) WT and Ccl17E/E mice (n = 5–6 mice/group) and (B) WT mice treated therapeutically with anti-CCL17 mAb or isotype control (150 μg i.p. on day 2 i.p.) (n = 4–5 mice/group). Pain and arthritis (histology) were measured. Original magnification, ×125. Results are shown as mean ± SEM. P values were obtained using a 2-way ANOVA test for pain and a 1-way (B) or 2-way (A) ANOVA test for histology quantification. *P < 0.05, **P < 0.01, ***P < 0.001, WT saline versus WT TNF; #P < 0.05, ###P < 0.001, WT TNF versus Ccl17E/E TNF; TNF + isotype versus TNF + anti-CCL17.

Article Snippet: Cells were treated with either murine GM-CSF (20 ng/ml) or PBS, for the indicated time periods, in the presence or absence of anti-TNF mAb (MP6-XT22, 1 ng/ml) (Schering BioPharma) ( 76 ) or IgG1 isotype control mAb (1 ng/ml).

Techniques: Methylation