Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 1. Interleukin-12 p70 secretion by lipopolysaccharide-stimulated adherent cells from the peripheral blood mononuclear cells of 27 burn and trauma patients measured by enzyme-linked immunoadsorbent assay on 53 occasions at serial intervals after injury and compared with 18 normal control subjects. The patients’ adherent cells produced significantly less interleukin-12 than cells from normal persons at multiple intervals beginning early after injury.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 2. Production of interleukin-12 (IL-12) and IL-10 by lipopolysaccharide-stimulated peripheral blood mononuclear cells (PBMCs) of burn and trauma patients at serial intervals after injury. Thirty-eight observations of IL-10 production were made in 18 patients and 21 observations of IL-12 production in 8 of the same patients. PBMC IL-12 production was significantly diminished in the 8- to 14-day interval after injury when compared with simultaneously studied normal subjects. At the same interval, IL-10 production by the patients’ PBMCs was significantly elevated.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques:

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 3. Survival of groups of 12 to 15 burn and sham burn mice after cecal ligation and puncture performed 10 days after injury. Burn mice receiving 5 ng interleukin-12 (IL-12) every other day for 9 days beginning on the day of injury had a survival similar to that of sham burn mice. IL-12 in a 10-ng dose appeared to have a less favorable effect on survival, and the addition of indomethacin to the 5-ng dose of IL-12 produced inferior results. IL-12 treatment did not affect the survival of sham burn mice.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 4. Survival of groups of 12 to 15 burn and sham burn mice after cecal ligation and puncture performed on day 10 after injury. The burn mice treated with 5 ng interleukin-12 (IL-12) every other day had a survival similar to that of sham burn controls, even though the IL-12 treatment was continued through day 11, beyond the time of cecal ligation and puncture. Untreated burn animals had the expected high death rate.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 5. Survival of groups of 12 to 15 burn and sham burn mice after cecal ligation and puncture performed on day 10 after injury. The dose of 1 ng interleukin-12 (IL-12) given every other day in burn animals and continued beyond the time of cecal ligation and puncture produced survival equivalent to that of sham burn controls.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 7. Interferon-gamma production by anti-CD3 antibody-stimulated splenocytes from groups of 5 to 10 burn and sham burn animals 10 days or 13 days after injury (after cecal ligation and puncture on day 10). The burn groups treated with interleukin-12 (IL-12) produced significantly more interferon-gamma than untreated animals. S and S/12 indicate untreated and IL-12–treated sham mice. B and B/12 indicate untreated and IL-12–treated burn mice.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 8. Tumor necrosis factor-alpha production by lipopolysaccharide-stimulated adherent splenocytes from groups of 5 to 10 burn and sham burn animals 10 days or 13 days after injury (after cecal ligation and puncture on day 10). Splenocyte production of tumor necrosis factor-alpha was markedly increased in the burn animals treated with interleukin-12.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 9. Production of interleukin-10 (IL-10) by anti-CD3–stimulated splenocytes harvested from groups of 5 to 10 burn and sham burn mice before cecal ligation and puncture on day 10 after injury or on day 13 (after cecal ligation and puncture on day 10). IL-12 treatment increased IL-10 production by burn splenocytes, both before and after cecal ligation and puncture. Splenocytes from untreated burn animals produced more IL-10 than sham burn splenocytes at both intervals.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 10. Total mononuclear cells per spleen in groups of five burn and sham burn animals at day 10 after the burn and at day 13, 3 days after cecal ligation and puncture was performed on day 10. The mononuclear cell population in the spleen was markedly reduced 3 days after cecal ligation and puncture, and treatment with interleukin-12 significantly increased the splenic mononuclear cell population of burn animals.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation

Journal: The Journal of Experimental Medicine

Article Title: A cyanobacterial LPS antagonist prevents endotoxin shock and blocks sustained TLR4 stimulation required for cytokine expression

doi: 10.1084/jem.20060136

Figure Lengend Snippet: CyP specifically inhibits LPS stimulation of DCs. (A) DCs were stimulated for 16 h with different TLR agonists (1 μg/ml LPS, 10 μg/ml PGN, 20 μg/ml poly(I:C), and 2.5 μg/ml R848), 20 ng/ml IL-1β, or 1 μg/ml soluble CD40L in the absence or presence of 20 μg/ml CyP. Data are expressed as the percentage of the response (TNF production, black bars; IL-6 production, white bars) obtained with the specific agonists in the absence of CyP and represent the mean ± SD of four independent experiments. Inhibition of LPS was found to be statistically significant (P < 0.0001), whereas inhibition of all other stimuli was found to be nonsignificant (P > 0.05). (B) DCs were challenged with graded numbers of DH5α bacteria in the absence (white bars) or presence (black bars) of 20 μg/ml CyP. TNF was measured in the 20-h culture supernatants by ELISA. CyP did not affect bacterial growth. One representative experiment of three is shown. (C) DCs were stimulated with LPS, 10 ng/ml IFN-γ, soluble CD40L, or R848 alone or in the indicated combinations in the absence (white bars) or presence (black bars) of CyP. IL-12p70 was measured in the 24-h culture supernatants. One representative experiment of four is shown.

Article Snippet: Cytokine production was measured in the supernatants of DCs (0.5 × 10 6 cells/ml) stimulated for 16–24 h using matched paired antibodies specific for human TNF, IL-6, and IL-12p70 (DuoSet ELISA development kits; R&D Systems).

Techniques: Inhibition, Bacteria, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Strain-Specific Human Natural Killer Cell Recognition of Influenza A Virus

doi: 10.1101/148528

Figure Lengend Snippet: (A) Cytokine concentrations assessed by Luminex®, values displayed represent the mean fluorescence intensity in pH1N1-infected/H3N2 infected conditions (MOI=3). Cytokines elevated by 2.5-fold over the level in mock-infected monocytes are plotted. (B) Impact of cytokine receptor blocking on the NK cell IFN-γ response evaluated by pre-incubating NK cells for 1 hr with blocking antibodies specific to IFNAR2, IL-12R, IL-15R, IL-18R, and IFNGR1 followed by co-culture with infected monocytes. 24 HPI, intracellular cytokine staining was used to assess NK cell IFN-γ + frequency compared to treatment with an isotype control antibody (H3N2: n =2-6, pH1N1: n =9). (C) NK cells were incubated for 1 hr with antibodies specific to IFNAR2 and CD226 or CD54 followed by co-culture with autologous infected monocytes. At 24 HPI, intracellular cytokine staining was used to assess IFN-γ production compared to treatment with an isotype control antibody ( n =6). * P < 0.05, ** P < 0.005, Wilcoxon signed-rank test. (D) Model of strain-specific NK cell recognition of influenza A infection. pH1N1-infection of monocytes virus does not downregulate CD54 and CD112 to the same extent as H3N2-infected monocytes. CD54 expression is retained on Flu-NP + cells while CD112 expression is preferentially retained on exposed, uninfected monocytes. pH1N1 infection of monocytes elicits enhanced IFN-α secretion and blockade of IFNAR2 dampens NK cell anti-pH1N1 IFN-γ production.

Article Snippet: IL-12R β 2 Ab (R&D Systems AF1959-SP) was used at a final concentration of 83.33 μg/mL.

Techniques: Luminex, Fluorescence, Infection, Blocking Assay, Co-Culture Assay, Staining, Control, Incubation, Virus, Expressing

Journal:

Article Title: Expression of the ?1?1 integrin, VLA-1, marks a distinct subset of human CD4 + memory T cells

doi: 10.1172/JCI200319607

Figure Lengend Snippet: VLA-1 expression is associated with Th1 polarization. (a) CD4+ PBLs were purified into VLA-1+ and VLA-1– fractions, activated with PMA/ionomycin for 5 hours, and analyzed for intracellular IFN-γ production. (b) CD4+ PBLs were stimulated with TT or anti-CD3; 10 days later the cells were harvested and purified into VLA-1+ and VLA-1– fractions, activated with PMA/ionomycin, and analyzed for intracellular IFN-γ and IL-4. (c) CD4+ PBLs were labeled with CFSE, stimulated with TT, and 10 days later purified into VLA-1+ and VLA-1– fractions. Subsequently, the two subsets were activated and analyzed for divisions and intracellular IFN-γ. (d) CD4+ PBLs were stimulated with TSST-1 in two different environments: Th1 (IL-12+) or TH2 (IL-4+ and anti–IFN-γ). Ten days later cells were harvested and analyzed for VLA-1 and Vβ2 surface expression (left) or activated and analyzed for intracellular IFN-γ (right). (e) Fresh SFLs from RA (n = 4) and PsA (n = 2) patients were immediately activated and analyzed for surface expression of CD4, VLA-1, and intracellular IFN-γ. The dot plots were obtained by pregating on CD4+ events, and a representative patient sample is shown.

Article Snippet: To induce Th1 polarization, the CD4 + cells were stimulated with 2.5 μg/ml of anti-CD3 mAb in a medium containing 5 ng/ml recombinant human IL-12 (R&D Systems Inc., Minneapolis, Minnesota, USA).

Techniques: Expressing, Purification, Labeling