Ginger extract inhibits LPS induced macrophage activation and function-1

<p><b>Copyright information:</b></p><p>Taken from "Ginger extract inhibits LPS induced macrophage activation and function"</p><p>http://www.biomedcentral.com/1472-6882/8/1</p><p>BMC Complementary and Alternative Medicine 2008;8():1-1.</p><p>Published online 3 Jan 2008</p><p>PMCID:PMC2234390.</p><p></p>with 100 ng/ml LPS in presence or absence of ginger extract for 24 h. The cells were harvested and stained with fluorophore conjugated mAb against B7.1 (), B7.2 () and CD40 () and analyzed in LSR II flow cytometer. Data are representative result of six replicates

Ginger extract inhibits LPS induced macrophage activation and function-4

<p><b>Copyright information:</b></p><p>Taken from "Ginger extract inhibits LPS induced macrophage activation and function"</p><p>http://www.biomedcentral.com/1472-6882/8/1</p><p>BMC Complementary and Alternative Medicine 2008;8():1-1.</p><p>Published online 3 Jan 2008</p><p>PMCID:PMC2234390.</p><p></p>ce of ginger extract for 24 h. These pre-treated macrophages were used as the sole source of APCs along with syngeneic T cells as the responder cells in the presence of allostimulation. IL-2 and IFN-γ production was measured in the culture supernates after 72 h. incubation period. * ≤ 0.05 in comparison to Control. # ≤ 0.05 in comparison to LPS

Ginger extract inhibits LPS induced macrophage activation and function-2

<p><b>Copyright information:</b></p><p>Taken from "Ginger extract inhibits LPS induced macrophage activation and function"</p><p>http://www.biomedcentral.com/1472-6882/8/1</p><p>BMC Complementary and Alternative Medicine 2008;8():1-1.</p><p>Published online 3 Jan 2008</p><p>PMCID:PMC2234390.</p><p></p>with 100 ng/ml LPS in presence or absence of ginger extract for 24 h. The cells were harvested and stained with fluorophore conjugated mAb against MHC II and analyzed in LSR II flow cytometer. Data are representative result of six replicates

Ginger extract inhibits LPS induced macrophage activation and function-3

<p><b>Copyright information:</b></p><p>Taken from "Ginger extract inhibits LPS induced macrophage activation and function"</p><p>http://www.biomedcentral.com/1472-6882/8/1</p><p>BMC Complementary and Alternative Medicine 2008;8():1-1.</p><p>Published online 3 Jan 2008</p><p>PMCID:PMC2234390.</p><p></p>ce of ginger extract for 24 h. These pre-treated macrophages were used as the sole source of APCs along with syngeneic T cells as the responder cells in the presence of allostimulation. T cell proliferation was measured by MTS assay after 72 h. incubation time. Results are presented as proliferation index equating the absorbance value (at 490 nm) of responder cells as 1. Responder cells only – no macrophages, Control- macrophages were not pre-treated, LPS- macrophages pre-treated with LPS only, LPS + G - macrophages pre-treated with LPS and ginger extract. Data represent mean ± S.D (n = 9). * ≤ 0.05 in comparison to Control. # ≤ 0.05 in comparison to LPS

Effect of RMT3-23 treatment on cytokine production by uNK cells.

<p>Treatment with RMT3-23 significantly down regulated cytokine production by NK1.1^- DX5^- DBA⁺ cell population. A significant decrease in the population of IFN, IL-6, IL-10, VEGF and GMCSF producing NK1.1^- DX5^- DBA⁺ cells was observed after treatment with RMT3-23 in comparison to Control group, whereas IL-4 production was significantly increased. Data are presented as mean± SEM and are representative of at least 3 experiments. Statistical analysis was done using unpaired t test.</p

Expression of CD11b, Ly6C and Ly6G on uNK cells from CBA mice.

<p>Female CBA mice were allogeneically mated toC57BL/6 males and uterine lymphocytes were isolated between GD 10.5 to 12.5. Expression of CD11b, Ly6C and Ly6G was determined on the NK1.1^- DX5^-DBA⁺ cell population. Data are representative of at least 4 experiments.</p

Expression of NK cell markers in the uNK and pNK population from non-pregnant and pregnant mice.

<p>Expression of NK1.1, DX5, DBA and NKp46 were determined on CD3^-CD122⁺ cells isolated from the uteri of non-pregnant (A) syngeneically mated pregnant (B) and allogeneically mated (C) pregnant CBA mice. Expression of NK1.1, DX5, DBA and NKp46 on CD3^-CD122⁺ cells from spleen of non-pregnant CBA mouse (D), syngeneically mated (E) and allogeneically mated (F) pregnant CBA mouse. Red-boxed areas denote uNK populations positive for NK1.1 and DBA in allogeneically mated pregnant CBA mouse (C) and DBA positive uNK population in syngeneically mated pregnant CBA mouse (B). These positive populations are absent in the splenic NK cell population in the same mouse. Data is representative of at least 3 experiments.</p

Effect of GM-CSF on the uNK cells and uMDSC from pregnant mice.

<p>A. CBA/CaJ female mice were mated with C57BL/6 allogeneic male mice. Uterine NK cells were flow sorted to obtain a NK1.1^- DX5^- DBA⁺ cell population that was incubated with GMCSF (20ng/ml) ± IL-2 (20ng/ml) for 5–7 days. Expression of uNK and CD11b⁺Ly6G^hiLy6C^lo and CD11b⁺Ly6G^loLy6C^hi cell populations were analyzed using a flow cytometer. GMCSF treatment resulted in no significant change in the NK1.1^- DX5^- DBA⁺ cell population from the CBA mice (p = 0.1164). However, an increase of the CD11b⁺Ly6G^loLy6C^hi population was observed (p = 0.0592). An inverse correlation was observed between the uNK cells and uMDSC in presence of both GMCSF and IL-2 treatment but was not statistically significant. B. C57BL/6 female mice were mated with allogeneic CBA/CaJ male mice. Uterine NK cells were flow sorted to obtain a NK1.1⁺ DX5^- DBA⁺ cell population that was incubated with GMCSF (20ng/ml) ± IL-2 (20ng/ml) for 5–7 days. Expression of uNK and CD11b⁺Ly6G^hiLy6C^lo and CD11b⁺Ly6G^loLy6C^hi cell populations were analyzed using a flow cytometer. GMCSF treatment resulted in a significant change in the NK1.1⁺ DX5^- DBA⁺ cell population from the C57BL/6 female mice (p = 0.0310). However, the change in the CD11b⁺Ly6G^loLy6C^hi population was not significant (p = 0.2410). The effect of GMCSF was more pronounced in the uNK cells from the C57BL/6 female mice and a significant inverse correlation between uNK and uMDSC was also observed (p = 0.0158). Data are presented as mean± SEM and are representative of at least 3 experiments. Statistical analysis was done by one-way analysis of variance (ANOVA) using Kruskal-Wallis test and two way ANOVA.</p

Effect of RMT3-23 treatment on expression of activation markers and inhibitory surface receptors on uNK cells.

<p>Treatment with RMT3-23 significantly upregulates the expression of CD69 on NK1.1^- DX5^- DBA⁺ cell population. A similar trend of increased expression was also observed in case of B220 and CD25 levels after treatment with RMT3-23 although the data were not statistically significant. RMT3-23 treatment also significantly down regulates Ly49C expression and up regulates KLRG1 and Ly49G2 on NK1.1^- DX5^- DBA⁺ cells. Data are presented as mean± SEM and are representative of at least 3 experiments. Statistical analysis was done using unpaired t test.</p

Effect of GM-CSF and IL-15 on the uNK cells from pregnant CBA mice.

<p>CBA/CaJ female mice were mated with C57BL/6 allogeneic male mice. Uterine lymphocytes were isolated from Control and RMT3-23 treated mice and cultured with IL-15, IL-15+IL-12, IL-12, IL-15+GMCSF and GMCSF only for 7days in vitro in a 37°C humidified CO2 incubator. At the end of the incubation period the percentage of 3 different cell populations were determined by flow cytometry. A. RMT3-23 treated uNK cells proliferated more in response to IL-12 (p = 0.0284). B. This effect was more pronounced in the uNK cells expressing the MDSC markers (p<0.0001).</p