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

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 1. Inhibition of allergic diarrhea disease by nasal treatment with IL-12p70 naked DNA plasmid. A, The incidence of allergic diarrhea was reduced in mice treated nasally with IL-12p70 DNA when compared with mice treated with empty vector plasmid. Allergic disease was induced in these mice by s.c. immunization and then repeated oral challenge (SC/ PO) with OVA. B, OVA-specific IgE Abs are reduced in the serum of allergic diarrhea-afflicted mice treated nasally with IL-12p70 DNA. C, To- tal IgE Abs are reduced in the serum of allergic diarrhea-afflicted mice treated with naked IL-12p70 DNA. The data are expressed as the mean SD and are representative of five independent experiments. Statistical dif- ferences between IL-12p70 DNA- and empty vector-treated mice (, p 0.01 and , p 0.05) are indicated.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Inhibition, Plasmid Preparation

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

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 2. Reduction of IgA and IgG hyperresponsiveness in large in- testinal B cells and of Th2 cells responses by nasal administration of IL- 12p70 DNA. A, The frequency of OVA-specific IgA, IgG, IgG1, and IgM AFCs in the large intestine of mice treated with IL-12p70 DNA or empty vector plasmid. B, OVA-specific IL-4 and IFN- production were analyzed by intracellular staining using FACS analysis. In vivo treatment with IL- 12p70 DNA reduced the predominant Th2-type Ag-specific responses by large intestinal mononuclear cells isolated from diarrhea-afflicted mice. The mononuclear cells isolated from the large intestine (1.0 106 cells/ well) or spleen were cultured with OVA (1 mg/ml) for 4 days. After in- cubation, cells were harvested and subjected to intracellular staining with anti-IL-4 and IFN-. The data are expressed as the percentage of cytokine- positive cells in large intestinal CD4 T cells. The data are expressed as the mean SD and are representative of four independent experiments. Sta- tistical differences between IL-12p70 DNA- and empty vector-treated mice (, p 0.01) are indicated. N.D., Not detected.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Plasmid Preparation, Staining, In Vivo, Isolation, Cell Culture

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

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 5. Induction of IL-12p70 protein in the large intestine by nasal administration of IL-12p70 DNA. An- ti-IL-12p70 mAb used did not cross- react with the IL-12p40 molecule. A, IL-12p70-positive cells were detected near large intestinal lymphatic vessels after nasal IL-12p70 DNA treatment. B, The IL-12p70 protein was also de- tected in large intestinal extracts using immunoprecipitation and blotting analysis. Recombinant murine IL- 12p70 (rIL-12p70) protein (lane 1) was used as a positive control for the detection of IL-12p70 (B). C, IL- 12p70-producing cells were stained with fluorescence-conjugated anti- CD11c in the large intestine.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Immunoprecipitation, Recombinant, Positive Control, Staining

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

Article Title: Nasal IL-12p70 DNA prevents and treats intestinal allergic diarrhea.

doi: 10.4049/jimmunol.174.11.7423

Figure Lengend Snippet: FIGURE 6. Nasal IL-12p70 DNA treatment cured allergic diarrhea. A, Frequency of allergic diarrhea de- creased after IL-12p70 DNA treat- ment (n 9 per group). B, The symp- toms of diarrhea were completely inhibited after three treatments with IL-12p70 DNA. C, Infiltration of eo- sinophils and basophils into the large intestine was blocked by the IL-12p70 DNA administration. The red arrows point to eosinophils and the blue ar- rows point to basophils. D, Results of the ELISPOT assay used to determine the OVA-specific Ig responses in the large intestinal mononuclear cells. The data are expressed as the mean SD and are representative of three in- dependent experiments. Statistical differences between IL-12p70 DNA- and empty vector-treated mice (, p 0.01) are indicated.

Article Snippet: After electrophoresis, proteins were transferred to a polyvinylidene difluoride microporous membrane (Immobilon; Millipore) and the membrane was reacted with biotinylated anti-IL-12p70 (48110.11, rat IgG1; R&D Systems) and then incubated with a biotin-streptavidin complex (ABC-AP kit; Vector Laboratories).

Techniques: Enzyme-linked Immunospot, Plasmid Preparation

Journal: Infection and Immunity

Article Title: Comparison of Pathogenesis and Host Immune Responses to Candida glabrata and Candida albicans in Systemically Infected Immunocompetent Mice

doi: 10.1128/iai.69.8.5046-5055.2001

Figure Lengend Snippet: FIG. 4. Temporal expression of TNF-a, IL-12, IFN-g, and IL-10 proteins in kidneys during C. glabrata and C. albicans infection. Crl:CF-1 mice were infected with virulent C. glabrata or C. albicans as described for Fig. 3. At specific time points p.i., the mice were euthanized, and kidneys were excised and homogenized. Levels of the immunoreactive cytokines TNF-a (a), IL-12 p70 (b), IFN-g (c), and IL-10 (d) were quantified in kidney homogenates. Results represent the mean 6 SEM of two separate experiments, 8 to 14 mice per treatment group. p, significantly greater than control mice, P , 0.05.

Article Snippet: Protein levels of TNF-a, IL-12, IL-10, and IFN-g were subsequently measured using commercially available cytokinespecific murine ELISA kits (Quantikine mouse TNF-a, mouse IL-12 p70, mouse IFN-g, and mouse IL-10; R&D systems, Minneapolis, Minn.) according to the manufacturer’s directions.

Techniques: Expressing, Infection, Control