Journal: Pathogens

Article Title: IFN-Gamma-Dependent and Independent Mechanisms of CD4 + Memory T Cell-Mediated Protection from Listeria Infection

doi: 10.3390/pathogens7010022

Figure Lengend Snippet: Heterologous challenge with Lm-gp61 induces increased levels of serum IFNγ. Naïve and LCMV-immune mice were challenged with Lm-gp61. Cytokine concentrations in the serum at day 1 and 3 post-infection were assessed using a cytokine bead array for ( A ) IFNγ; ( B ) IL-12p70; ( C ) TNF; ( D ) IL-1β; ( E ) IL-10; and ( F ) IL-6. n = 8 mice/group, data are pooled from two separate experiments. Dotted lines indicate limit of detection for the assay.

Article Snippet: 0.5 mg anti-TNF neutralizing antibodies (BioXCell, XT3.11) or anti-IL-6 (BioXCell, MP5-20F3) were given i.p. 1 day prior to infection and then every other day after that until sacrifice.

Techniques: Infection

Journal: Pathogens

Article Title: IFN-Gamma-Dependent and Independent Mechanisms of CD4 + Memory T Cell-Mediated Protection from Listeria Infection

doi: 10.3390/pathogens7010022

Figure Lengend Snippet: Neutralization of TNF prevents accumulation of activated macrophages in the spleen. Naïve and LCMV-immune mice were challenged with Lm-gp61 and some groups of mice were additionally treated with neutralizing antibodies to IFNγ or TNF. ( A ) Splenocytes were gated for F4/80 + CD11b + macrophages. The bar graph indicates the frequency of macrophages expressing IFNγR1 (CD119) for each treatment group. Accompanying flow plots indicate representative gating for CD119 + macrophages, as compared to isotype control. Frequencies were obtained by subtracting the percentage that were stained by the isotype control; ( B ) The bar graph indicates the change in MFI of CD119 staining on F4/80 + CD11b + macrophages for each treatment group. The change in MFI was obtained by subtracting the MFI following isotype control staining from the MFI following CD119 staining. The accompanying flow plot indicates representative staining for CD119 on macrophages for each treatment group; ( C ) RNA from FACS-sorted F4/80 + CD11b + macrophages was analyzed by semi-quantitative RT-PCR for changes in cytokine transcript levels. Bar plots indicate the relative fold change in expression between treatment groups for the indicated transcripts. n = 3–5 mice per group.

Article Snippet: 0.5 mg anti-TNF neutralizing antibodies (BioXCell, XT3.11) or anti-IL-6 (BioXCell, MP5-20F3) were given i.p. 1 day prior to infection and then every other day after that until sacrifice.

Techniques: Neutralization, Expressing, Staining, Quantitative RT-PCR

Journal: Pathogens

Article Title: IFN-Gamma-Dependent and Independent Mechanisms of CD4 + Memory T Cell-Mediated Protection from Listeria Infection

doi: 10.3390/pathogens7010022

Figure Lengend Snippet: Neutralization of IFNγ and TNF alter the inflammatory environment induced by secondary challenge. Naïve and LCMV-immune mice were challenged with Lm-gp61 and some groups of mice were additionally treated with neutralizing antibodies to IFNγ or TNF. Bar plots show the concentration of ( A ) IFNγ; ( B ) TNF and ( C ) IL-6 in the serum of the indicated treatment groups at day 1 or 3 after Lm-gp61 challenge. n = 7–8 mice per group, data are representative of two separate experiments.

Article Snippet: 0.5 mg anti-TNF neutralizing antibodies (BioXCell, XT3.11) or anti-IL-6 (BioXCell, MP5-20F3) were given i.p. 1 day prior to infection and then every other day after that until sacrifice.

Techniques: Neutralization, Concentration Assay

Journal: Journal of Neuroinflammation

Article Title: Mesenchymal stem cells exert anti-proliferative effect on lipopolysaccharide-stimulated BV2 microglia by reducing tumour necrosis factor-α levels

doi: 10.1186/s12974-014-0149-8

Figure Lengend Snippet: Mesenchymal stem cells modulate the expression of IL-6 and TNF-α in microglia co-culture. Microglia (BV2 cells or primary microglia) and Mesenchymal stem cells (MSC) were seeded simultaneously into 24-well plates at a 1:0.2 ratio. Cells were left overnight in culture and stimulated with 1 μg/ml lipopolysaccharide (LPS). Culture supernatants were collected 24 hours later and expression of (a,b) interleukin (IL)-6 and (c,d) tumour necrosis factor (TNF)-α were analysed using the RayBio antibody array (a,c) or BD Cytokine Bead Array (b,d). Protein array results are expressed as mean relative protein level ± standard deviation from two independent experiments and cytokine bead array results are expressed in mean concentration (pg/ml) ± standard deviation from three independent experiments. * P < 0.001, versus BV2 cells; # P < 0.001, versus BV2 + LPS.

Article Snippet: Blocking antibodies against TNF-α (XT3.11), IL-6 (MP5-20 F3) and isotype (Rat IgG1) control (all from Bio X Cell, NH, USA) were reconstituted in 1X PBS to 1.0 mg/ml.

Techniques: Expressing, Co-Culture Assay, Ab Array, Protein Array, Standard Deviation, Concentration Assay

Journal: Journal of Neuroinflammation

Article Title: Mesenchymal stem cells exert anti-proliferative effect on lipopolysaccharide-stimulated BV2 microglia by reducing tumour necrosis factor-α levels

doi: 10.1186/s12974-014-0149-8

Figure Lengend Snippet: Nitric oxide surge in co-culture and reduction in microglial proliferation are mediated by TNF-α modulation. BV2 cells and mesenchymal stem cells (MSC) were seeded at a 0.2 ratio into 96 well plates. After overnight incubation, 1 μg/ml lipopolysaccharide (LPS) and (a,c) tumour necrosis factor (TNF)-α blocking antibodies or (b,d) recombinant TNF-α were added at concentrations indicated below the graph, and NO 2 - concentration (a,b) and proliferation (c,d) was analysed at 48 hours. Values are expressed as mean ± standard deviation from three independent experiments. Controls are culture wells without blocking antibodies. * P < 0.05, versus control BV2 cells; # P < 0.05, ## P < 0.001, versus control BV2 + LPS; ф P < 0.05, фф P < 0.001, versus control co-culture + LPS.

Article Snippet: Blocking antibodies against TNF-α (XT3.11), IL-6 (MP5-20 F3) and isotype (Rat IgG1) control (all from Bio X Cell, NH, USA) were reconstituted in 1X PBS to 1.0 mg/ml.

Techniques: Co-Culture Assay, Incubation, Blocking Assay, Recombinant, Concentration Assay, Standard Deviation, Control