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Abstract
Effective microorganism (EM) fermentation extract has been widely used for agricultural and environmental application. It has been recently revealed that EM cocktail treatment may be effective for treatment of diseases including cancer. In the present study, effectiveness of EM cocktail to control asthma was investigated using a mouse model of allergic asthma. Asthmatic mice sensitized and intranasally challenged with OVA were orally given EM fermentate (EM-1®) during antigen challenge. Administration of EM-1® resulted in a significant reduction in airway hyper-reactivity (AHR) and airway recruitment of total leukocytes and eosinophils. Cytokine (IL-4, IL-5 and IFNγ) levels in bronchoalveolar lavage fluid (BALF) and lung tissues were not altered by EM-1® treatment. However, IL-13 level in BALF was considerably lower in EM-1® treated mice than in controls. Moreover, Ag-specific IL-4, IL-5 and IL-13 production of draining lymph node cells were markedly downregulated by EM-1® treatment when compared to controls, whereas their IFNγ production was not significantly different. Those data show that EM-1® treatment suppresses type 2 helper T (Th2), but not type 1 helper T (Th1), cell response. This finding was also supported by serum antibody data showing that IgE and IgG1 levels in EM-1® treated mice were significantly lower than in controls, while IgG2a level was not significantly different between two groups. In conclusion, oral administration of EM-1® attenuates asthmatic manifestations including AHR and airway recruitment of eosinophils in a mouse model and which possibly results from selective inhibition of Th2 cell response to allergen. Our data also suggest that EM-1® may be effectively applied for control of allergic asthma.
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Figure 1.
Effect of oral administration of EM-1® on airway hyperreactivity in a mouse model of asthma. Mice were sensitized with OVA at days 0 and 7, and then challenged with aerosolized OVA from day 15 to day 20 and EM-1® was orally given from day 14 for 7 consecutive days. One day after last challenge, Penh was determined in response to increasing doses of methacholine. Data are means ±SE of 10 mice from 3 separate experiments. ∗, p<0.05, ∗∗, p<0.005 vs each control.
Figure 2.
Effect of oral administration of EM-1® on airway inflammation in a mouse model of asthma. Mice were immunized and EM-1® was orally given as described in Materials and Methods. After examination of AHR, the trachea was cannulated and the lungs were lavaged. Total number of live leukocytes in recovered BALF (A) was enumerated (p=0.008, n=7), and differential cell counts (B) were performed on Diff Quik-stained cytocentrifuge preparations. ∗p=0.015; ∗∗p=0.001 vs control (n=7).
Figure 3.
Effect of oral administration of EM-1® on cytokine levels in inflammatory sites. Mice were immunized and EM-1® was orally given as described in Materials and Methods. Levels of cytokines in BALF and homogenized lung tissues were determined by ELISA. ∗, p=0.012 vs control (n=6).
Figure 4.
Effect of oral administration of EM-1® on Ag-specific responses of lymph node cells. One day after last challenge, peribronchial LN cells were prepared and stimulated with OVA. At day 3 of culture, number of viable cells (A) and cytokine production (B) were examined. ∗∗p<0.005 vs each control (n=6).
Figure 5.
Effect of oral administration of EM-1® on Ag-specific antibody responses in vivo. After examination of airway lavage, blood was taken by cardiac puncture and serum was recovered. Ag-specific serum IgE (A), IgG1 (B) and IgG2a (C) levels was evaluated by ELISA. ∗, p<0.05; ∗∗, p<0.01 vs each control (n=6).
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