TWI381843B - Food composition and pharmaceutical composition for treating allergic thermo lethal strains - Google Patents

Description

Food composition and pharmaceutical composition for treating allergic thermo lethal strains

The present invention relates to a food composition and a pharmaceutical composition, and more particularly to a food composition and a pharmaceutical composition comprising an immunologically active thermo lethal strain for treating allergy.

Generally, products containing lactic acid bacteria (LAB) only have the health effect of regulating the intestinal tract. Although there are tens of thousands of strains of lactic acid bacteria in nature, only a few lactic acid strains have anti-allergic properties. The ability of a few of these bacterial strains to have acid and bile salt tolerance, ability to adsorb mucosal epithelial cells, and ability to survive after passage through the gastrointestinal tract is an important basis for screening for strains that promote health effects. To date, only a few strains of lactic acid bacteria that have been proven to have anti-allergic health effects have been identified, and the function of lactic acid bacteria for health is in the nature of strains rather than species. "Guidelines for the evaluation of probiotics in food; Report of joint FAO/WHO working group on drafting guidelines for the evaluation of probiotics in food; London Ontario, Canada April 30 And May 1, 2002: 1-7).

The relationship between probiotics and immune response has been concentrated since the beginning of 2000 and has been extensively studied until today. Most of the probiotics are Gram-positive bacteria. After entering the body from outside, they are attached to the mucosal cells of the human body and bind to the terpenoid receptors of dendritic cells and macrophages. The substances recognized by each type of sputum receptor are different, except that the steroid receptors on the cell surface are more easily identified as polysaccharide substances, such as peptidoglycan or lipopolysaccharide, and intracellular steroid receptors. Only to identify substances in the body, such as cytoplasmic proteins or genetic material DNA strands. The substances contained in the cells react with the terpenoid receptors to mature the dendritic cells or macrophages and secrete related cytokines to stimulate the immune response. At present, it is found that the most typical phenomenon of probiotics affecting immune response is to stimulate the activation of type 1 helper T cells, and the second phenomenon is the maturation of type 1 regulatory T cells. These two different pathways have been distinguished by the cytokines that are stimulated by dendritic cells or macrophages. In the former, the cytokines secreted by the two antigen-presenting cells are mostly proinflammatory cytokines such as IL-12, IFN-γ, TNF-α, etc.; the cytokines secreted by the latter are mostly anti-inflammatory cytokines such as IL- 10.

There are many literature reports on lactic acid bacteria related to allergic reactions at home and abroad. In an animal test study on the ability of lactic acid bacteria to reduce allergies, animal experiments by the heat-killing lactic acid bacteria L. plantarum L-137 can induce IL-12 and IFN-γ production by macrophages and spleen cells of DBA/2 mice. It also inhibits IgE with casein specificity and IgG1 antibody specific for casein (Murosaki, 1998). Feeding L. casei Shirota around OVA-TCR-Tg mice increased spleen cells secretion of IFN-γ and IL-12, and decreased secretion of IL-4 and IL-5 (Shida et al., 2002). OVA-sensitized BALB/c mice were fed with Lactobacillus fermented milk for 27 days to reduce OVA-specific IgE in serum (Ishida et al., 2003). C3H/Hej mice are sensitized with OVA and cholera toxin, and B. bifidum BGN4 and L. casei 911 can reduce OVA-specific IgE, IgA, total IgE, and IgG1 in serum (Kim et al., 2005), these documents confirm that in animal models of allergic patterns, feeding lactic acid bacteria can reduce the level of IgE in the serum, thereby improving the allergy.

Lactobacillus in clinical trials, patients with allergic diseases can really improve allergic reactions, combined with dry lactic acid bacteria L. rhamnosus and L. reuteri for six weeks, is beneficial for allergic diseases, questionnaires for parents to assess probiotics for one to thirteen years old Children with atopic dermatitis found an improvement in atopic dermatitis in 56% of children (Rosenfeldt et al., 2003). Ishida (2005) scholars for the treatment of forty-nine patients with allergic rhinitis, drinking L. acidophilus strain L-92, also found that allergic symptoms have improved. Eight weeks after taking L. fermentum VRI 003 PCCTM, fifty-three children with atopic dermatitis can increase IFN-γ production in PBMC cells and improve allergic conditions (Prescott et al., 2005). F Lster-Holst (2006) and other scholars have pointed out that the test of 54 patients with atopic dermatitis can effectively improve allergy symptoms after taking L. rhamnosus strain GG and placebo.

Live lactic acid bacteria with anti-allergic properties must be stored by refrigerating or freezing in order to prevent their anti-allergic activity from being lowered by storage at room temperature for a long time. The immunosuppressive heat-killing strain can solve the problem of the general anti-allergic probiotic product stored at room temperature after being killed by heat at a specific temperature.

In one aspect, the invention broadly comprises a composition comprising the entero faecalis YM-73 strain, the Food Industry Development Institute, accession number BCRC 910442, and a physiologically acceptable excipient or diluent.

In another specific embodiment, the physiologically acceptable homologous or mutant species of the heat-killing Enterococcus faecium strain YM-73 is enhanced by the Food Industry Development Institute Registry Number BCRC 910442.

In another embodiment, the invention broadly comprises inhibiting immunoglobulin IgE by promoting Th1 type immune response by promoting intercellular white pigment (IL-12) or interferon gamma, and improving Th2 type immune response excess. Allergic phenomenon, which is a strain of Y. faecalis YM-73 which can be administered to mammals to stimulate the dose of Th1 type immunity, and the Food Industry Development Institute registration number BCRC 910442.

The invention may also be broadly construed as including a part or a combination of elements and features in the description of the application, and any or all combinations of any one or more of those parts, constituent elements and features. And where well-known and equivalents are described in the art to which the present invention pertains, such known equivalents will be incorporated herein.

The present invention broadly includes the heat-killing Enterococcus faecalis YM-73 strain, Food Industry Development Institute, registration number BCRC 910442.

Many literatures have pointed out that increasing the secretion of first-type T-cell cytokines, including interleukin (IL-12) and interferon-gamma (IFN-gamma), can effectively improve allergy symptoms; further studies have pointed out specific lactic acid bacteria strains and trees. The binding of Toll-like receptors on the spheroid cells, the transfer of the transgenic proteins in the activated cells to the nucleus and the release of a large number of cytokines, is a part of innate immunity, so some specific lactic acid bacteria species have more cell walls. Carbohydrates such as peptidoglycan, polysaccharide, etc., through the innate immune system, can indeed activate the development of T cells in the body.

The freeze-dried culture of the strain of the present invention has been deposited at the Taiwan Food Industry Development Research Institute at 331 Food Road, Hsinchu City, Republic of China. The details of the deposit are shown in Table 1:

The deposited strains listed in Table 1 have been found to have anti-allergic ability after heat-killing under appropriate conditions, including slowing and treatment of atopic dermatitis, urticaria, allergic rhinitis, food allergies and asthma symptoms. Features.

Example 1: Morphology and general properties of the strains of the invention.

The taxonomic characteristics of the strains were confirmed based on the 16S rDNA sequence analysis and the results of the API bacterial identification system analysis. The strain of the present invention was found to be Enterococcus faecalis. The morphological and general characteristics of this strain are detailed in Table 2:

Example 2: The effect of the strain of the present invention on promoting Th1 type immunity by promoting secretion of interferon γ after heat-killing under appropriate conditions (using peripheral blood mononuclear cells as an in vitro efficacy verification platform).

The effect of the strain of the present invention, Enterococcus faecalis YM-73 strain BCRC 910442, on the immunity of Th1 type is measured, which is a method for measuring human peripheral blood mononuclear cells (PBMC) and the present invention. The strains were co-cultured and the amount of interferon γ secreted was measured to verify the strain having anti-allergic ability. Use the following experimental steps:

1. Take an appropriate human blood volume of about 200 mL of whole blood each time.

2. Take an equal ratio of Ficoll-paque and blood at 180-20g for 30-40 minutes at 18-20 °C.

3. Take the white blood cell layer of human peripheral blood, wash the cells 2-3 times with a buffer solution, and suspend the cells with a suitable medium (for example, RPMI-1640).

4. After the strain of the present invention which has been activated for 3 days is thermally killed by the temperature of 55 ° C to 80 ° C and 90 ° C to 120 ° C, the live bacteria and the cells which are heat-killed under different conditions are respectively ratio of 1:10. Cultivate with cells for 48 hours.

5. Collect the supernatant from the cell culture. The content of interferon gamma (IFN-gamma) in the supernatant was determined by enzyme immunoassay (ELISA).

Statistical analysis of the data is shown in Table 3 and Figure 1, expressed as mean ± standard deviation (Mean ± SD). As shown in FIG 10, respectively ⁶ to 10 ⁸ cfu of E. faecalis strain YM-73 BCRC viable treatment conditions and two kinds of thermal death of the cells 910,442 and ¹⁰⁵ to ¹⁰⁷ individual category 1 peripheral blood monocytes The cells (PBMC) were co-cultured for 48 hours, and the cell supernatant was collected, and the content of interferon gamma (IFN-gamma) in the supernatant was detected by enzyme immunoassay. Among them, the experimental background value (Cell only) was used as the negative control group, and the bacteria that reported the anti-allergic lactic acid bacteria ( Lactobacillus rhamnosus GG BCRC 16000) and the cells treated with the same heat lethal condition were functional control groups. PHA (Phytohemagglutinin) is a positive control group that detects the concentration of interferon gamma stimulated. The results showed that the two heat-killing conditions of Enterococcus faecalis YM-73 cells significantly stimulated the secretion of interferon-gamma from human peripheral blood mononuclear cells (PBMC) and the control group ( Lactobacillus rhamnosus GG BCRC 16000). At a heat lethal temperature of 55 ° C to 80 ° C, the fascination of Enterococcus faecalis YM-73 was 36 times higher than that of the control group ( Lactobacillus rhamnosus GG BCRC 16000); at the temperature of 90 ° C to 120 ° C, the faecal intestine The bacillus YM-73 cells were 15 times more potent than the control group ( Lactobacillus rhamnosus GG BCRC 16000) (PBMC). Table 3 shows the active cells of the Enterococcus faecalis YM-73 strain and the control strain, the heat lethal condition bacteria at 55 ° C to 80 ° C, the heat lethal condition cells at 90 ° C to 120 ° C, and the control group and the peripheral blood of the human Culture of mononuclear cells (PBMC) stimulates the secretion of interferon gamma (means ± SD):

Example 3: The effect of the strain of the present invention on promoting the secretion of interleukin 12p70 (IL-12p70) to regulate Th1 type immunity (using peripheral blood mononuclear cells as an in vitro efficacy verification platform).

To examine the effect of the strain of the present invention, Enterococcus faecalis YM-73 strain BCRC 910442, on the immunity of Th1 type, which is determined by co-culture of human peripheral blood mononuclear cells (PBMC) with the strain of the present invention. The amount of interleukin 12p70 (IL-12p70) secreted to verify the strain with anti-allergic ability. Use the following experimental steps:

1. Take an appropriate human blood volume of about 200 mL of whole blood each time.

2. Take an equal ratio of Ficoll-paque and blood at 180-20g for 30-40 minutes at 18-20 °C.

3. Take the white blood cell layer of human peripheral blood, wash the cells 2-3 times with a buffer solution, and suspend the cells with a suitable medium (for example, RPMI-1640).

4. After the strain of the present invention which has been activated for 3 days is thermally killed by the temperature of 55 ° C to 80 ° C and 90 ° C to 120 ° C, the live bacteria and the cells which are heat-killed under different conditions are respectively ratio of 1:10. Cultivate with cells for 48 hours.

5. Collect the supernatant from the cell culture. The content of intercellular white 12p70 (IL-12p70) in the supernatant was determined by enzyme immunoassay (ELISA).

The statistical analysis of the data is shown in Table 4 and Figure 2, and is expressed in Mean ± SD. Figure 2 shows when the heat is killed by 10⁶ To 10⁸ Live bacteria of C. faecalis YM-73 strain BCRC 910442 and cells treated with two heat lethal conditions and 10⁵ To 10⁷ Individual peripheral blood mononuclear cells (PBMC) were cultured for 48 hours, cell supernatants were collected, and the content of interleukin 12p70 (IL-12p70) in the supernatant was detected by enzyme immunoassay. Among them, the experimental background value (Cell only) was used as the negative control group, and most of the literature was published to prove that it has anti-allergic lactic acid bacteria (Lactobacillus rhamnosus The viable bacteria treated by GG BCRC 16000 and the cells treated with the same heat lethal condition were the functional control group, and PHA (Phytohemagglutinin) was the positive control group, and the stimulated concentration of interleukin 12p70 (IL-12p70) was detected. The results showed that the two heat-killing conditions of Enterococcus faecalis strain YM-73 could significantly stimulate the secretion of intercellular white pigment 12p70 from human peripheral blood mononuclear cells (PBMC) and the control group (Lactobacillus rhamnosus GG BCRC 16000) has a significant difference. Under the heat lethal temperature of 55 ° C ~ 80 ° C, Enterococcus faecalis YM-73 cells compared to the control group (Lactobacillus rhamnosus The stimulation amount of GG BCRC 16000) is 50 times higher; the thermogenic lethal temperature of 90 °C ~ 120 °C, the Enterococcus faecalis YM-73 cells are better than the control group (Lactobacillus rhamnosus The stimulating amount of GG BCRC 16000) is 18 times higher (PBMC). Table 4 is the active bacteria of the strain and the control strain of the present invention, the heat lethal condition bacteria at 55 ° C to 80 ° C, the heat lethal condition cells at 90 ° C to 120 ° C, and the control group and the peripheral blood singles of the human Culture of nucleated cells (PBMC) stimulates the secretion of interleukin 12p70 (means±SD):

Example 4: The effect of the strain of the present invention on reducing the secretion of interleukin 13 (IL-13) to regulate the immunity of Th2 type (using peripheral blood mononuclear cells as an in vitro efficacy verification platform).

The inhibitory effect of the strain of the present invention, Enterococcus faecalis YM-73 strain BCRC 910442, on the immunity of Th2 type, which is determined by co-culture of human peripheral blood mononuclear cells (PBMC) with the strain of the present invention. The amount of interleukin 13 (IL-13) secreted to verify the strain with a Th2-type immune response to counteract allergic abilities. Use the following experimental steps:

1. Take an appropriate human blood volume of about 200 mL of whole blood each time.

2. Take an equal ratio of Ficoll-paque and blood at 180-20g for 30-40 minutes at 18-20 °C.

3. Take the white blood cell layer of human peripheral blood, wash the cells 2-3 times with a buffer solution, and suspend the cells with a suitable medium (for example, RPMI-1640).

4. After the strain of the present invention which has been activated for 3 days is thermally killed by the temperature of 55 ° C to 80 ° C and 90 ° C to 120 ° C, the live bacteria and the cells which are heat-killed under different conditions are respectively ratio of 1:10. Cultivate with cells for 48 hours.

5. Collect the supernatant from the cell culture. The content of intercellular white 13 (IL-13) in the supernatant was determined by enzyme immunoassay (ELISA).

The statistical analysis of the data is shown in Table 5 and Figure 3, and is expressed in Mean ± SD. As shown in FIG 10, respectively ⁶ to 10 ⁸ cfu of E. faecalis strain YM-73 BCRC viable treatment conditions and two kinds of thermal death of the cells 910,442 and ¹⁰⁵ to ¹⁰⁷ individual category 3 peripheral blood monocytes The cells (PBMC) were co-cultured for 48 hours, and the supernatant of the cells was collected, and the content of interleukin 13 (IL-13) in the supernatant was detected by enzyme immunoassay. Among them, the experimental background value (Cell only) was used as the negative control group, and the bacteria that reported the anti-allergic lactic acid bacteria ( Lactobacillus rhamnosus GG BCRC 16000) and the cells treated with the same heat lethal condition were functional control groups. PHA (Phytohemagglutinin) is a positive control group that detects the concentration of interleukin 13 (IL-13) stimulated. The results showed that the two heat-killing conditions of Enterococcus faecalis strain YM-73 could significantly stimulate the peripheral blood mononuclear sphere (PBMC) to reduce the secretion of intercellular white pigment 13 and the positive control group (PHA). Under the heat lethal temperature of 55 °C ~ 80 °C, the stimuli of Enterococcus faecalis YM-73 strain was 6 times lower than that of the control group (Lactobacillus rhamnosus GG BCRC 16000); the Enterococcus faecalis YM was found at the heat lethal temperature of 90 °C ~ 120 °C. The stimuli of the -73 strain were also nearly 6-fold lower than the control group (Lactobacillus rhamnosus GG BCRC 16000) (PBMC). Table 4 is the active bacteria of the strain and the control strain of the present invention, the heat lethal condition bacteria at 55 ° C to 80 ° C, the heat lethal condition cells at 90 ° C to 120 ° C, and the control group and the peripheral blood singles of the human Nucleation of nucleus cells (PBMC) stimulates the secretion of interleukin 13 (means ± SD):

Example 5: A live-type scanning electron microscope (FE-SEM) of a living cell of the strain of the present invention and two appropriate conditions of heat lethal temperature were used to observe the cell conformation test.

Since the state of heat lethality may cause some damage to the cells or changes in the configuration of the material outside the cell wall, a field emission scanning electron microscope (FE-SEM) was used to amplify the state of the cells by 50,000 times, and the state of the cells was observed. In the case of heating and letting of bacteria, changes in the configuration of external substances, such as increased surface wrinkles, may cause the cell wall polysaccharides of the cells to increase in contact with the recipients of immune cells, or the destruction of the cells, releasing Protein bodies or genetic material inside the cells, such as DNA, enhance the ability of these substances to stimulate immune cells. The active ingredient of the strain of the present invention may be derived from the polysaccharide on the cell wall, the cytoplasmic protein or glycoprotein in the cell, the lipoprotein component, or the genetic material deoxyribonucleic acid (DNA), thus heating the cell The effect is precisely the release of the active ingredient of the anti-allergic strain, and the anti-allergic effect of the active bacteria is more intense.

The results of field-scanning scanning electron microscopy (FE-SEM) of Enterococcus faecalis YM-73 strain BCRC 910442 are shown in Fig. 4. Figure 4A shows the heat-killed bacteria freeze-dried into powder at 55 °C ~ 80 °C, and is magnified 50,000 times under FE-SEM; B is frozen at 60 °C ~ 120 °C. The powder was dried and magnified at 50,000 times under FE-SEM. In the C-picture, the active cells were freeze-dried into a bacterial powder and magnified 50,000 times under FE-SEM. From the comparison of A and C, it can be found that after heat-killing the strain at 55 °C to 80 °C, the wrinkles on the surface of the strain become more and the cells are more concentrated; and the comparison between B and C shows that The strain was destroyed and there was a broken cell. Therefore, at these two temperatures, the ability of the cells to stimulate the immune cells to produce Th1 cytokines is about 4.6 times stronger than that of the living cells.

Example 6: Test for the adsorption capacity of the strain of the present invention on human intestinal epithelial cells (Caco-2).

The ability of the human intestinal epithelial cells (Caco-2) to be adsorbed by the active cells of the strain of the present invention and the heat-killing cells of the two suitable conditions are analyzed in vitro using the differentiated cell strain. The human intestinal epithelial cell (Caco-2) monolayer cell strain was first grown on a cover glass to a monolayer, and then placed in a multi-well cell culture dish. Then, the ratio of the cells: the strain of the present invention is 1:200, and the cells are cultured for 1-3 hours. After washing with a buffer solution and fixing the cells with methanol and the remaining number of adhering bacteria, the gram stain is determined and the attached is determined. The number of bacteria. Referring to the analysis method of Jacobsen et al. (1999), 20 fields of view were counted under a 1000-fold microscope, and when the average number of bacteria per field of view was less than 40, it was judged as nonadhesive; when the average number of bacteria per field of view When it is between 41 and 100, it is judged to be adherent; when the average number of bacteria per field of view is more than 100, it is judged to be strongly adhesive.

Statistical analysis of the data is expressed in Mean ± SD. On average, the calculated values of forty-five fields of view are taken, and the results are summarized in Table 6 and Figure 5. Fig. 5 shows that the active cells of Enterococcus faecalis YM-73 strain BCRC 910442 and the cells after heat-killing at 55 ° C to 80 ° C were judged as "attachment"; the cells after heat death at 90 ° C to 120 ° C were judged as " Not attached." Table 6 shows the adsorption capacity of the active cells of Enterococcus faecalis YM-73 strain BCRC 910442 and the heat-killing bacteria of two appropriate conditions on human intestinal epithelial cells (Caco-2) cell line:

Because probiotics have to exert anti-allergic effects, in addition to finding strains with specific functions, they also have good adsorption capacity for small intestinal mucosal epithelial cells. Also because of this property, the strain of the present invention can provide a medical use for treating or soothing allergy symptoms. The strain of the present invention can simultaneously enhance the Th1 pathway and simultaneously regulate an allergic reaction that inhibits Th2 hypersaturation. It is an object of the present invention to continue to achieve these desires or at least to provide the public with a new choice of steroids or anti-tissue ammonia in allergy treatment. The present invention finds allergic lactic acid bacteria which have no side effects and are beneficial to the human body as an allergy treatment. New choice.

The embodiments described above are only intended to illustrate the technical idea and the features of the present invention, and the purpose of the present invention is to enable those skilled in the art to understand the contents of the present invention and to implement the present invention. That is, the equivalent variations or modifications made by the spirit of the present invention should still be included in the scope of the present invention.

Figure 1 shows peripheral cells treated with 10 ⁶ to 10 ⁸ cfu of Enterococcus faecalis YM-73 strain BCRC 910442 and two heat-killing conditions, and 10 ⁵ to 10 ⁷ human peripheral blood mononuclear cells. (PBMC) were co-cultured for 48 hours, cell supernatant was collected, and the content of interferon gamma (IFN-gamma) in the supernatant was detected by enzyme immunoassay. Among them, the experimental background value (Cell only) was used as the negative control group, and the bacteria that reported the anti-allergic lactic acid bacteria ( Lactobacillus rhamnosus GG BCRC 16000) and the cells treated with the same heat lethal condition were functional control groups. PHA (Phytohemagglutinin) is a positive control group that detects the concentration of interferon gamma stimulated. The results showed that the two heat-killing conditions of Enterococcus faecalis strain YM-73 significantly stimulated the secretion of interferon-gamma from human peripheral blood mononuclear cells (PBMC) and the control group ( Lactobacillus rhamnosus GG BCRC 16000). At a heat lethal temperature of 55 ° C to 80 ° C, the strain of Enterococcus faecalis YM-73 was 36 times more irritating than the control group ( Lactobacillus rhamnosus GG BCRC 16000); the Enterococcus faecalis at a temperature of 90 ° C to 120 ° C The YM-73 strain was 15 times more potent than the control group ( Lactobacillus rhamnosus GG BCRC 16000) (PBMC).

Figure 2 shows the peripheral blood mononuclear cells of 10 ⁵ to 10 ⁷ humans when they were killed by heat-killing 10 ⁶ to 10 ⁸ cfu of Enterococcus faecalis YM-73 strain BCRC 910442 and two heat-killing conditions, respectively. The globular cells (PBMC) were co-cultured for 48 hours, and the supernatant of the cells was collected, and the content of the interleukin 12p70 (IL-12p70) in the supernatant was detected by enzyme immunoassay. Among them, the experimental background value (Cell only) was used as the negative control group, and the bacteria that reported the anti-allergic lactic acid bacteria ( Lactobacillus rhamnosus GG BCRC 16000) and the cells treated with the same heat lethal condition were functional control groups. PHA (Phytohemagglutinin) was used as a positive control group to detect the stimulated concentration of interleukin 12p70 (IL-12p70). The results showed that the two heat-killing conditions of Enterococcus faecalis strain YM-73 could significantly stimulate the peripheral blood mononuclear sphere (PBMC) secreting intercellular albinin 12p70 and the control group ( Lactobacillus rhamnosus GG BCRC 16000). At a heat lethal temperature of 55 ° C to 80 ° C, the strain of Enterococcus faecalis YM-73 was 50 times more irritating than the control group ( Lactobacillus rhamnosus GG BCRC 16000); the enterococci were found at a heat lethal temperature of 90 ° C to 120 ° C. The YM-73 strain was 18 times more potent than the control group ( Lactobacillus rhamnosus GG BCRC 16000).

Figure 3 shows the peripheral blood mononuclear cells of 10 ⁵ to 10 ⁷ humans when they were killed by heat-killing 10 ⁶ to 10 ⁸ cfu of Enterococcus faecalis YM-73 strain BCRC 910442 and two heat-killing conditions, respectively. The globular cells (PBMC) were co-cultured for 48 hours, and the supernatant of the cells was collected, and the content of interleukin 13 (IL-13) in the supernatant was detected by enzyme immunoassay. Among them, the experimental background value (Cell only) was used as the negative control group, and the bacteria that reported the anti-allergic lactic acid bacteria ( Lactobacillus rhamnosus GG BCRC 16000) and the cells treated with the same heat lethal condition were functional control groups. PHA (Phytohemagglutinin) is a positive control group that detects the concentration of interleukin 13 (IL-13) stimulated. The results showed that the two heat-killing conditions of Enterococcus faecalis strain YM-73 could significantly stimulate the peripheral blood mononuclear sphere (PBMC) to reduce the secretion of intercellular white pigment 13 and the positive control group (PHA). Under the heat lethal temperature of 55 °C ~ 80 °C, the stimuli of Enterococcus faecalis YM-73 strain was 6 times lower than that of the control group ( Lactobacillus rhamnosus GG BCRC 16000); the Enterococcus faecalis YM under the heat lethal temperature of 90 °C ~ 120 °C The amount of -73 strain was also nearly six times lower than that of the control group ( Lactobacillus rhamnosus GG BCRC 16000).

Figure 4 shows that A is freeze-dried into a bacterial powder of Enterococcus faecalis YM-73 at 55 °C to 80 °C, and is magnified 50,000 times under FE-SEM; B is hot at 90 °C to 120 °C. The lethal faecal strain YM-73 was freeze-dried into a bacterial powder and magnified 50,000 times under FE-SEM. C is the active cell of the Enterococcus faecalis YM-73 strain freeze-dried into a bacterial powder, which was amplified by FE-SEM. 50000 times observed. From the comparison of A and C, it can be found that after heat-induced Enterococcus faecalis YM-73 strain at 55 °C ~ 80 °C, the surface of the strain becomes more wrinkled and the cells become more concentrated; and B and C In comparison, it was found that the Enterococcus faecalis strain YM-73 was destroyed and had broken cells. Therefore, at these two heat lethal temperatures, the ability of the cells to stimulate the immune cells to produce Th1 cytokines is about 4.6 times stronger than that of the active cells.

Figure 5 shows that according to the analysis method of Jacobsen et al. (1999), when the average number of bacteria per field of view is less than 40, it is judged as nonadhesive; when the average number of bacteria per field of view is between 41 and 100 It is judged to be adherent; when the average number of bacteria per field of view is more than 100, it is judged to be strongly adhesive. The active cells showing the Enterococcus faecalis YM-73 strain BCRC 910442 and the cells after heat-killing at 55 ° C to 80 ° C were judged as "adhered"; the cells after heat death at 90 ° C to 120 ° C were judged as "not attached". "."

Claims (8)

A food composition which inhibits immunoglobulin IgE by regulating secretion of intercellular melanin (IL-12) or interferon gamma to regulate Th1 type immune response, thereby improving an allergic phenomenon of Th2 type immune reaction excess, the food composition Including: Enterococcus faecalis YM-73, which stimulates immune cells to secrete anti-allergic-related cytokines, Taiwan Hsinchu Food Industry Development Institute, accession number BCRC 910442; and physiologically acceptable excipients or diluents. The food composition of claim 1, wherein the heat lethal temperature is 55 ° C to 80 ° C. The food composition of claim 1, wherein the heat lethal temperature is from 90 ° C to 120 ° C. The food composition of claim 1, wherein the excipient or diluent is a food product. The food composition of claim 4, wherein the food product comprises fermented milk, yogurt, cheese, milk powder for dairy beverages, tea, coffee or a combination thereof. A pharmaceutical composition for treating allergy, which inhibits immunoglobulin IgE by promoting secretion of intercellular melanin (IL-12) or interferon gamma to regulate Th1 type immune response, thereby improving allergy of Th2 type immune reaction The pharmaceutical composition comprises: Enterococcus faecalis YM-73, which stimulates immune cells to secrete anti-allergic-related cytokines, Taiwan Hsinchu Food Industry Development Institute, accession number BCRC 910442; and pharmaceutically acceptable form Agent or diluent. The pharmaceutical composition for treating allergy according to claim 6, wherein the heat lethal temperature is 55 ° C to 80 ° C. The pharmaceutical composition for treating allergy according to claim 6, wherein the heat lethal temperature is from 90 ° C to 120 ° C.