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1.
Figure 5

Figure 5. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

IL-12 production by DC or purified peritoneal macrophages stimulated with an extract of A. blazei or ABPC. (a) DCs were coincubated with an extract of A. blazei (0·1%) or ABPC (0·05%) for 72 hr. Cell free culture supernatants were then collected and IL-12 p40 was measured by ELISA. (b) Purified peritoneal macrophages were coincubated with en extract of A. blazei (open square) or ABPC (open circle) at the indicated concentrations for 72 hr. Cell free culture supernatants were then collected and IL-12 p40 was measured by ELISA. Data are shown as mean ± SD of triplicate samples. Similar results were obtained in three independent experiments.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.
2.
Figure 2

Figure 2. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

MNC numbers and NK cell percentage after A. blazei oral administration for 2 weeks. Two weeks after daily oral administration of A. blazei suspension (32 mg/500 µl/head) into WT B6 mice, spleen and liver MNCs number (a) and populations of liver MNC were analyzed with flow cytometry (b). Data are shown as mean ±SD of three to five mice in each group. Similar results were obtained in two independent experiments.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.
3.
Figure 6

Figure 6. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

IL-12- and IL-18-dependent IFN-γ induction by A. blazei. (a) DC-rich spleen MNCs isolated from B6 mice were coincubated with an extract of A. blazei or ABPC at the indicated concentration in the presence (open circle) or absence (open square) of anti-IL-12 mAb for 72 hr. Cell free culture supernatants were then collected and IFN-γ was measured by ELISA. (b) DC-rich spleen MNCs isolated from B6 mice were coincubated with an extract of A. blazei at the indicated concentration in the presence of anti-IL-12 mAb (open circle), anti-IL-18 mAb (open triangle), anti-IL-12 mAb and anti-IL-18 mAb (open diamond), or control immunoglobulins (open square) for 72 hr. Cell free culture supernatants were then collected and IFN-γ was measured by ELISA. Data are shown as mean ± SD of triplicate samples. Similar results were obtained in three independent experiments.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.
4.
Figure 3

Figure 3. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

IFN-γ dependent NK cell activation by oral administration of A. blazei. WT, RAG-2–/–, and IFN-γ–/– B6 mice were orally administered daily with A. blazei (32 mg/500 µl/head) (open circle) or water (500 µl/head) (open square) for 2 weeks. Then, NK cells were purified from liver MNCs and cytotoxic activity was analysed using NK cell-sensitive target cells, YAC-1, at indicated E/T ratios. Data are shown as mean ± SD of triplicate samples. Similar results were obtained in three independent experiments. *P < 0·05.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.
5.
Figure 1

Figure 1. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

Activation of NK cell cytotoxicity by oral administration with A. blazei. (a) B6 mice were administered daily with A. blazei suspension (32 mg/500 µl/head) (open circle) or water (500 µl/head) (open square) for 1 or 2 weeks. Then, liver MNCs were prepared and cytotoxic activity was analysed using NK cell-sensitive target cells, YAC-1, at indicated E/T ratios. (b) B6 mice were administered daily with 500 µl of suspension containing 0 mg (open square), 16 mg (open diamond), 32 mg (open circle), or 64 mg (open triangle) of A. blazei for 2 weeks. Then, liver MNCs were prepared and cytotoxic activity was analysed using NK cell-sensitive target cells, YAC-1. (c) C3H/HeJ and BALB/c mice were administered daily with A. blazei suspension (32 mg/500 µl/head) (open circle) or water (500 µl/head) (open square) for 2 weeks. Then, liver MNCs were prepared and cytotoxic activity was analysed using NK cell-sensitive target cells, YAC-1, at indicated E/T ratios. Data are shown as mean ± SD of triplicate samples. Similar results were obtained in two or three independent experiments. *P < 0·05.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.
6.
Figure 4

Figure 4. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

IFN-γ induction in vitro by an extract of A. blazei or ABPC. (a) DC-rich spleen MNCs isolated from B6 or C3H/HeJ mice were coincubated with an extract of A. blazei (open square) or ABPC (open circle) at the indicated concentrations for 72 hr. Cell free culture supernatants were then collected and IFN-γ was measured by ELISA. (b) LPS independent IFN-γ induction. DC-rich spleen MNCs isolated from B6 or C3H/HeJ mice were coincubated with an extract of A. blazei or ABPC in the presence (10 µg/ml) (open circle) or absence (open square) of polymixin B for 72 hr. Cell free culture supernatants were then collected and IFN-γ was measured by ELISA. (c) Sufficient inhibition by polymixin B on IFN-γ production induced by LPS. DC-rich spleen MNCs isolated from B6 or C3H/HeJ mice were coincubated with LPS (5 µg/ml) in the presence (10 µg/ml) or absence of polymixin B for 72 hr. Cell free culture supernatants were then collected and IFN-γ was measured by ELISA. (d) Major role of NK cells in IFN-γ production. NK cell-depleted or intact DC-rich spleen MNCs isolated from RAG-2–/– B6 mice were coincubated with an extract of A. blazei (0·1%) or ABPC (0·05%) for 72 hr. Cell free culture supernatants were then collected and IFN-γ was measured by ELISA. Data are shown as mean ± SD of triplicate samples. Similar results were obtained in two or three independent experiments.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.
7.
Figure 8

Figure 8. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

IL-12-mediated activation of NK cell cytotoxicity induced by an extract of A. blazei or ABPC in vivo. (a) WT B6 mice were administered daily with A. blazei extract (500 µl/head) (square), ABPC extract (250 µl/head)(circle) or water (500 µl/head) (triangle) for 2 weeks. Some mice were also administrated with 300 µg of anti-IL-12 mAb (closed) or control immunoglobulin (open) every 3 days. Then, liver MNCs were prepared and cytotoxic activity was analysed using NK cell-sensitive target cells, YAC-1, at the indicated E/T ratios. *P < 0·05 when the cytotoxic activity is compared with that of control MNCs from water-administered mice. (b) IFN-γ–/– B6 mice were administered daily with A. blazei extract (500 µl/head) (square), ABPC extract (250 µl/head) (circle) or water (500 µl/head) (triangle) for 2 weeks. Then, liver MNCs were prepared and cytotoxic activity was analysed using NK cell-sensitive target cells, YAC-1, at the indicated E/T ratios. Data are shown as mean ± SD of triplicate samples. Similar results were obtained in two or three independent experiments.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.
8.
Figure 7

Figure 7. From: Interleukin-12- and interferon-?-mediated natural killer cell activation by Agaricus blazei Murill.

IL-12 mediated IFN-γ-dependent NK cell cytotoxicity induced by an extract of A. blazei or ABPC in vitro. (a) Augmentation of NK cell cytotoxicity by an extract of A. blazei or ABPC. DC-rich spleen MNC isolated from B6 mice were cultured with an extract of A.blazei (0·1%) (open triangle), ABPC (0·05%) (open circle), or control water (open square) for 72 hr. Cells were then harvested and cytotoxic activity was examined using the NK cell-sensitive target, YAC-1, at the indicated ratios. *P < 0·05. (b) IFN-γ dependent augmentation of cytotoxicty by an extract of A. blazei or ABPC. DC-rich spleen MNC from WT (square) or IFN-γ–/– (circle) B6 mice were cultured in the presence (closed) or absence (open) of an extract of A.blazei (0·1%) or ABPC (0·05%) for 72 hr. Cells were then harvested and cytotoxic activity was examined. *P < 0·05 when the cytotoxic activity of stimulated MNC is compared with that of control MNCs. (c) IL-12-dependent augmentation of NK cell cytotoxicty by an extract of A. blazei or ABPC. DC-rich spleen MNCs isolated from B6 mice were cultured in the presence (closed) or absence (open) of an extract of A.blazei (0·1%) or ABPC (0·05%) with anti-IL-12 mAb (circle) or control immunoglobulin (square). After 72 hr incubation, cells were harvested and cytotoxic activity was analysed. In some experiments, NK cells were depleted from DC-rich MNC stimulated without anti-IL-12 mAb before cytotoxic assay (triangle). *P < 0·05 when the cytotoxic activity is compared with that of control MNC. Data are shown as mean ± SD of triplicate samples. Similar results were obtained in two or three independent experiments.

Eri Yuminamochi, et al. Immunology. 2007 June;121(2):197-206.

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