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

Article Title: Selective suppression of IL-12 production by chemoattractants.

doi: 10.4049/jimmunol.164.6.3009

Figure Lengend Snippet: FIGURE 1. IL-12 p70 production from elutri- ated human monocytes stimulated with SAC (0.01%) and IFN-g (100 ng/ml) in the presence or the absence of MCP-1 to -4 or C5a (100 nM). Values are means of duplicate assays for IL-12 p70 production from monocytes from four indi- vidual donors.

Article Snippet: ELISA reagents for human IL-12 p70 were purchased from R&D Systems and for IL-10 and TNF-a from BioSource International (Camarillo, CA); the respective capture and detection Abs were as follows: IL-12 p70, clone 24945.11; and polyclonal goat IgG, IL-10 clones AHC8102 and AHC7109, and TNF-a clones AHC3712 and AHC3419.

Techniques:

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

Article Title: Selective suppression of IL-12 production by chemoattractants.

doi: 10.4049/jimmunol.164.6.3009

Figure Lengend Snippet: FIGURE 3. Production of IL-12 p70 by human monocytes following SAC (0.01%) and IFN-g (100 ng/ml) stimulation in the presence of varying concentrations of either MCP-1 or C5a. Data are the mean 6 SD from three separate donors (p, p , 0.05; pp, p , 0.01).

Article Snippet: ELISA reagents for human IL-12 p70 were purchased from R&D Systems and for IL-10 and TNF-a from BioSource International (Camarillo, CA); the respective capture and detection Abs were as follows: IL-12 p70, clone 24945.11; and polyclonal goat IgG, IL-10 clones AHC8102 and AHC7109, and TNF-a clones AHC3712 and AHC3419.

Techniques:

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

Article Title: Selective suppression of IL-12 production by chemoattractants.

doi: 10.4049/jimmunol.164.6.3009

Figure Lengend Snippet: FIGURE 5. IL-12 p70 production from monocytes stimulated with SAC (0.01%) and IFN-g (100 ng/ml) with or without preexposure to MCP-1 to -4 or C5a (100 nM). Monocytes were cultured under the following condi- tions: neutralizing Ab to IL-10 or isotype-matched control Ab (A), neu- tralizing Ab to TGF-b1 or control Ab (B), and with or without indometh- acin (C). Data are the mean 6 SD from three separate donors (p . 0.05 for all variables tested).

Article Snippet: ELISA reagents for human IL-12 p70 were purchased from R&D Systems and for IL-10 and TNF-a from BioSource International (Camarillo, CA); the respective capture and detection Abs were as follows: IL-12 p70, clone 24945.11; and polyclonal goat IgG, IL-10 clones AHC8102 and AHC7109, and TNF-a clones AHC3712 and AHC3419.

Techniques: Cell Culture, Control

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

Article Title: Selective suppression of IL-12 production by chemoattractants.

doi: 10.4049/jimmunol.164.6.3009

Figure Lengend Snippet: FIGURE 6. Effect of pertussis toxin on chemoattractant-mediated sup- pression of IL-12 p70. Monocytes were cultured for 2 h with 250 ng/ml of pertussis toxin and then stimulated with SAC (0.01%) and IFN-g (100 ng/ml) in the presence or the absence of MCP-1 to -4 or C5a (100 nM). Data are the mean 6 SD from three separate donors (p, p , 0.05; pp, p , 0.01).

Article Snippet: ELISA reagents for human IL-12 p70 were purchased from R&D Systems and for IL-10 and TNF-a from BioSource International (Camarillo, CA); the respective capture and detection Abs were as follows: IL-12 p70, clone 24945.11; and polyclonal goat IgG, IL-10 clones AHC8102 and AHC7109, and TNF-a clones AHC3712 and AHC3419.

Techniques: Cell Culture

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

Article Title: Selective suppression of IL-12 production by chemoattractants.

doi: 10.4049/jimmunol.164.6.3009

Figure Lengend Snippet: FIGURE 7. Effects of chemoattractants on IL-12 p70 production from monocyte-derived DC. Immature DC (CD1ahigh/CD83low) were stimulated with either SAC (0.01%) and IFN-g (100 ng/ml) or CD40L (3 mg/ml) and IFN-g (100 ng/ml) in the presence or the absence or various chemoattrac- tants (100 nM). The figure is representative data from one of two experi- ments using DC derived from different donors.

Article Snippet: ELISA reagents for human IL-12 p70 were purchased from R&D Systems and for IL-10 and TNF-a from BioSource International (Camarillo, CA); the respective capture and detection Abs were as follows: IL-12 p70, clone 24945.11; and polyclonal goat IgG, IL-10 clones AHC8102 and AHC7109, and TNF-a clones AHC3712 and AHC3419.

Techniques: Derivative Assay

Journal: The Journal of Experimental Medicine

Article Title: Interferon-α and Interleukin-12 Are Induced Differentially by Toll-like Receptor 7 Ligands in Human Blood Dendritic Cell Subsets

doi: 10.1084/jem.20020207

Figure Lengend Snippet: TLR-7 ligands induce the production of IL-12 and IFN-α from MDCs and PDCs, respectively. (A and B) After 24-h culture with various stimuli, concentration of IL-12 p40+p70 (A) and IFN-α (B) in the culture supernatants of MDCs, PDCs, and PBMCs were measured by ELISA. The data are shown by means ± SEM of five independent experiments. (C) After 5-h culture with R-848 (10 −6 M), intracellular staining of IL-12 and IFN-α in MDCs and PDCs, together with the staining of surface expression of CD40, was performed. Percentages of the respective cytokine producing DCs are indicated. This figure represents the results from one of three experiments.

Article Snippet: 10 μg/ml brefeldin A (Sigma-Aldrich) was added during the last 1 h. After the stimulation, MDCs and PDCs were stained with Cy-Chrome-labeled CD40 (5C3; BD PharMingen), and then fixed, permeabilized (FIX and PERM kit; Caltag Laboratories), and stained with FITC-labeled anti–IL-12 p40+p70 mAb (B-P24; DIACLONE Research) or unconjugated mouse anti–human IFN-α mAb (MC-16; Genzyme).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Monophosphoryl lipid A plus IFNγ maturation of dendritic cells induces antigen-specific CD8 + cytotoxic T cells with high cytolytic potential

doi: 10.1007/s00262-010-0843-z

Figure Lengend Snippet: The superior induction of CTLs by MPLA/IFNγ DCs compared to CC-DCs is dependent on IL-12 and not on IL-6 or IL-23. a Blocking antibodies specific for IL-12p40 and IL-6, and an irrelevant antibody were added to MPLA/IFNγ DC-CTL cocultures. 7 days after the second stimulation of the CTLs with DCs (day 17), the CTLs were stained with the MART-1 tetramer (TM) and CD8-FITC. A graph showing the % MART-1 TM cells for all separate cultures of one experiment is shown. The condition treated with anti-IL-12 significantly differed (one-way ANOVA, P < 0.05) from untreated or isotype control treated conditions. A representative experiment out of four is depicted. b Blocking antibodies specific for IL-12p35 and IL-23p19, and an irrelevant antibody were added to MPLA/IFNγ DC-CTL cocultures. 7 days after the third stimulation of the CTLs, the CTLs were stained with the MART-1 TM and CD8-FITC. A graph showing the % MART-1 TM cells for all separate cultures of one experiment is shown. The condition treated with anti-IL-12p35 significantly differed (one-way ANOVA, P < 0.01) from isotype control treated condition

Article Snippet: The following neutralizing antibodies were used; anti-IL-12p40 (C8.6), anti-IL-12p35 (B-T21, Diaclone), anti-IL-23p19 (B-Z23, Diaclone), anti-IL-6 (IL6.8 [ 26 ]).

Techniques: Blocking Assay, Staining, Control

Journal: NPJ Vaccines

Article Title: Toxoplasma WH3 Δ rop18 acts as a live attenuated vaccine against acute and chronic toxoplasmosis

doi: 10.1038/s41541-024-00996-9

Figure Lengend Snippet: Detection of cytokines and T. gondii -specific IgG level in the sera of mice at 30/75/125 dpv with WH3 Δ rop18 by ELISA test. a Strategy diagram of the experimental process. b IFN-γ, c IL-12p70, d TNF-α, e IL-10 ( n = 5) and f Toxoplasma -specific total IgG and IgG1 and IgG2a ( n = 8). Serum was derived from unvaccinated mice as a negative control. *** P < 0.001, ** P < 0.01, * P < 0.05, ns not significant.

Article Snippet: In the meantime, the expression levels of cytokines interferon-gamma (IFN-γ), interleukin 12p70 (IL-12p70), tumor necrosis factor TNF (TNF-α) and interleukin 10 (IL-10) were detected by using ELISA kits (ProteinTech Group, Inc., USA) according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Negative Control

Journal: NPJ Vaccines

Article Title: Toxoplasma WH3 Δ rop18 acts as a live attenuated vaccine against acute and chronic toxoplasmosis

doi: 10.1038/s41541-024-00996-9

Figure Lengend Snippet: a Schematic illustration of the experimental procedure. b , d Percentage of CD3 + CD8 + T cells and CD3 + CD4 + T cells detected with FCM ( n = 4). c , e Increased percentage of Th1 (CD4 + IFN-γ + ) cell population in the mouse spleens following WH3 Δ rop18 immunization compared to the control ( n = 4). f Spleen cells cultured in vitro were stimulated with 10 µg/mL WH3 WT soluble antigens. Subsequently, the levels of IFN-γ, TNF-α, IL-12p70 and IL-10 in the splenic cell culture supernatant were detected by ELISA ( n = 5). Unpaired t -tests were used for statistical analysis. Bars = mean ± SEM. * P < 0.05, *** P < 0.001.

Article Snippet: In the meantime, the expression levels of cytokines interferon-gamma (IFN-γ), interleukin 12p70 (IL-12p70), tumor necrosis factor TNF (TNF-α) and interleukin 10 (IL-10) were detected by using ELISA kits (ProteinTech Group, Inc., USA) according to the manufacturer’s recommendations.

Techniques: Control, Cell Culture, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: NPJ Vaccines

Article Title: Toxoplasma WH3 Δ rop18 acts as a live attenuated vaccine against acute and chronic toxoplasmosis

doi: 10.1038/s41541-024-00996-9

Figure Lengend Snippet: 10 3 WH3 Δ rop18 parasites were i.p. into BALB/c mice for 75 days as a vaccine dose. Then, the mice were challenged with 10 3 WH3 WT or 10 5 WH6 tachyzoites. Unvaccinated mice were treated similarly. a Schematic illustration of the study design. b The vaccinated mice were challenged with 10 3 WH3 WT tachyzoites or c 10 5 WH6 tachyzoites. The survival rate was recorded for 35 days ( n = 10). Gehan-Breslow-Wilcoxon tests. d WH3 WT or WH6 tachyzoites were used for challenges of vaccinated mice, and 7 days later, the parasite burden in ascites were tested by qPCR ( n = 5). *** P < 0.001. e IFN-γ, IL-12p70, TNF-α and IL-10 in sera were tested by ELISA ( n = 5), *** P < 0.001. f , g Toxoplasma -specific total IgG and subclasses of IgG1 and IgG2a in sera were tested by ELISA. Unpaired t -tests were used for s t atistical analysis. Bars = mean ± SEM. *** P < 0.001.

Article Snippet: In the meantime, the expression levels of cytokines interferon-gamma (IFN-γ), interleukin 12p70 (IL-12p70), tumor necrosis factor TNF (TNF-α) and interleukin 10 (IL-10) were detected by using ELISA kits (ProteinTech Group, Inc., USA) according to the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: NPJ Vaccines

Article Title: Toxoplasma WH3 Δ rop18 acts as a live attenuated vaccine against acute and chronic toxoplasmosis

doi: 10.1038/s41541-024-00996-9

Figure Lengend Snippet: 10 3 WH3 Δ rop18 parasites were i.p. into BALB/c mice as a vaccine dose and observed for 125 days. Vaccinated or un-vaccinated mice were then infected with 10 3 RH or 10 5 ME49 parasites. a Schematic illustration of the study design. b 10 3 RH or c 10 5 ME49 tachyzoites were used for challenges ( n = 10). The survival rate was recorded for 35 days. Gehan-Breslow-Wilcoxon tests. d RH or ME49 tachyzoites challenges in vaccinated mice, followed by detection of parasite burden by qPCR in the ascites of animals 7 days later ( n = 5). *** P < 0.001. e IFN-γ, IL-12p70, TNF-α and IL-10 in sera were tested by ELISA ( n = 5), *** P < 0.001, ** P < 0.01. f , g Toxoplasma -specific total IgG and subclasses of IgG1 and IgG2a in sera were tested by ELISA ( n = 10). Unpaired t -tests were used for statistical analysis. Bars = mean ± SEM. *** P < 0.001.

Article Snippet: In the meantime, the expression levels of cytokines interferon-gamma (IFN-γ), interleukin 12p70 (IL-12p70), tumor necrosis factor TNF (TNF-α) and interleukin 10 (IL-10) were detected by using ELISA kits (ProteinTech Group, Inc., USA) according to the manufacturer’s recommendations.

Techniques: Infection, Enzyme-linked Immunosorbent Assay