JP3057300B2 - Composition for suppressing decline of immune function with aging - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a composition for suppressing a decrease in immune function. In general, as one of the aging phenomena, the so-called "senile disease", which is likely to develop due to a decrease in immune function, requires prevention or treatment, and therefore the present invention relates to such aging. The present invention relates to an oral composition for suppressing the accompanying decrease in immune function. [0002] In humans and experimental animals, the amount of lymphatic tissue in the thymus decreases with age due to atrophy of the thymus cortex.
It is said that T cell production and lymphocyte redistribution to T cell regions of lymph nodes are impaired. [0003] Therefore, as the aging progresses, a disease called so-called geriatric disease tends to develop. [0004] Methods for suppressing the decline of the immune function with aging and restoring the immune function of elderly individuals include regulating the nutritional status, administering thymic hormones and chemicals, and transplanting cells. For example, it has been reported that feeding a calorie-restricted diet to rats increases their lifespan. Examples of administration of thymic hormone include thymosin and thymopoietin. [0005] Examples of immunostimulants include double-stranded polynucleotides and sulfhydryl compounds. Also, a method of transplanting young lymphocytes or tissues into old individuals has been attempted. [0006] The method of feeding a calorie-restricted diet to rats cannot be applied to humans, because a restricted diet during the human growth phase suppresses the development of immune functions. Because. Thymic hormone alone is not capable of inducing mature T cells from bone marrow stem cells by itself, and none of them can completely substitute for thymic function. Oral administration is also not possible due to the problem of side effects. Immunostimulants are still at the research stage. At present, transplantation of young lymphocytes or tissues into old individuals cannot be performed easily. Accordingly, an object of the present invention is to provide a composition for suppressing a decrease in immune function, which prevents or treats a disease that is likely to develop due to a decrease in immune function with aging. [0008] The present invention means to solve the above-mentioned object, immune dysfunction suppressing composition with age comprising the milk milked from sensitized cows with bacterial antigens as active ingredients (where mixing of the gastrointestinal tract
Prevention and treatment of rheumatoid arthritis caused by bacterial infection
Excluding) . The antigen used in the present invention may be pathogenic or non-pathogenic as long as it is derived from a microorganism. The present inventors have sensitized cows with one or more bacterial antigens selected from the group of microorganisms shown in Table 2 below (hereinafter abbreviated as "S-100 antigen" for convenience). Milking from dairy cows sensitized by booster administration of the antigen several times (hereinafter, for convenience, "immune milk")
This was administered to mice. [Table 2] Also, Japanese Patent Application Laid-Open Nos.
Milk milked from cows to which the antigen disclosed in JP 1188523 has been boosted has the same effect. As such milk, cows are sensitized by administering a bacterial vaccine, and then a sufficient amount of the same antigen as the sensitizing bacteria is administered,
A sensitized one is desirable. On the other hand, for comparison with this "immune milk", commercially available cow's milk (hereinafter abbreviated as "control milk" for convenience) was administered to mice. Next, for each of the above mice,
The following immunological indices were measured, and their changes were observed over time. Mixed lymphocyte culture reaction of spleen Mixed lymph node culture reaction of mesenteric lymph node Mitogen responsiveness of mesenteric lymph node As a result of these measurements, immune milk was more controllable It was found that both activities increased more than milk. Based on this finding, the present inventors can prevent or treat aging-related diseases by administering immunized milk, that is, the so-called “separate medicinal effect” of immunized milk (new use).
I found Conventionally, this type of immune milk has been used for rheumatoid joints.
It was only used to prevent and treat inflammation. Arthritis
Prevention and prevention of immune dysfunction are completely different concepts.
You. On the basis of the above-mentioned novel facts, the present inventors have found a composition containing immune milk as an active ingredient that suppresses the deterioration of immune function with aging, and completed the present invention. The composition of the present invention for suppressing the deterioration of immune function with aging can be administered in the form of milk powder, liquid milk, yogurt or tablets. Next, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to only this embodiment. Example 1 Ten cows were immunized with a multivalent vaccine made with antigens containing all of the bacterial antigens in Table 2 above. [0022] In this case, the immunization method used was 5 ml of a vaccine containing 4 × 10 ⁸ cells / ml of each heat-sterilized cell per week.
Sensitize by administering booster for 4 consecutive weeks at the rate of
The primary immunization was performed. Then, the antibody titer of each cow to the antigen was confirmed by the agglutination method. Thereafter, the amount of the cells was boosted several times. The cows immunized in this manner were milked daily to obtain immunized milk. The immunized milk was processed as necessary, for example, after defatting, and then sterilized and spray-dried under temperature control to obtain powdered milk. Next, an experimental example will be described. The milk used is as follows. Immune milk: Immunized milk (milk powder) obtained in Example 1 Control milk: Normal commercially available milk (milk powder) Experiment 1: Mixed lymphocyte culture reaction of spleen Method: 8 weeks Age C57BL / 6CrSLC
Fourteen female mice were divided into two groups, and each group (7 mice) was mixed with 48.9 g of powder of immunized milk or control milk in 11.1 g of the mixed feed and continuously fed at 3 g / animal / day. For 61-63 weeks. Thereafter, the animals were sacrificed, and the removed spleen was added to a petri dish containing 10 cc of RPMI 1640, crushed with two slide glasses, and the released cells were collected and washed. On the other hand, allogeneic C3H or BA
LB / C mice were sacrificed and the removed spleens were irradiated with a radiation dose of 25 Gy, and the resulting spleen cells were used as stimulator cells. Using the spleen cells of the above 69-71 week old mouse (hereinafter referred to as “aged mouse”), 5 × 10 ⁵ / well of each spleen cell of C3H or BALB / C mouse 5 × 10 ⁵ / well And cultured at 37 ° C. for 4 days in a CO ₂ incubator. Four hours before the end of the culture, 1 μCi (37 MBq) well of ³ H thymidine was added, and after the end of the culture, the cells were collected on a filter mat, and the amount of tritiated thymidine incorporated in the cells was measured with a β counter. And the relative stimulation index (Specifici) of the mixed lymphocyte culture reaction of the spleen of the aged mouse.
cSI) was calculated. Then, the mixed lymphocyte culture reaction of C57BL / 6CrSLC female mice (hereinafter, referred to as "young mice") of 7 to 9 weeks old, which is different from the aging mice, was performed in the same manner as described above. The relative stimulation index was calculated, and the change of the relative stimulation index of the aged mice to which the immunized milk or the control milk was given was compared based on the relative stimulation index of the young mouse. Table 3 shows the results. [Table 3] Next, for each of the cases where C3H and BALB / C are used as indices, the value obtained by subtracting the value of the relative stimulation index of the aged mouse (B) from the value of the relative stimulation index of the young mouse (A). (C) was compared with the group to which the immunized milk or the control milk was administered to examine how much the group to which the immunized milk suppressed the aging-related decrease in the spleen mixed lymphocyte culture reaction due to aging. Table 4 shows the results. [Table 4] When the results were examined, the degree of the decrease in the mixed lymphocyte culture reaction of the spleen, which decreased with age, was as follows:
It was confirmed that the group using the immunized milk significantly suppressed compared to the group using the control milk. The mixed lymphocyte reaction was carried out based on Shigeru Muramatsu, “Experimental Physiology Course”, Vol. 14, Immunobiology, 1985, pp. 383-385. The relative stimulation index is represented by the following equation (I). ## EQU1 ## Experiment 2: Mixed lymphocyte culture reaction of mesenteric lymph nodes Method: 8-week-old C57BL / 6CrSLC
Fourteen female mice were divided into two groups, and each group (7) was mixed with 48.9 g of powder of immunized milk or control milk and mixed feed 1
1.1 g, 3 g / animal per day
The animals were allowed to ingest for 9-71 weeks, then sacrificed, the mesenteric lymph nodes removed by opening the abdomen and 10 cc of RPMI 16
Place in a plate containing 40 media. 2 in the scene
Crush with two slide glasses. The released cells were collected by filtration with a gauze, and lymphocytes of mesenteric lymph nodes were obtained. On the other hand, allogeneic C3H or BA
LB / C mice were sacrificed and spleen cells obtained after irradiating the isolated spleen with a radiation dose of 25 Gy were used as stimulator cells. Using lymphocytes contained in the mesenteric lymph node of the aged mouse, 5 × 10 ⁵ / well of 5 × 10 ⁵ spleen cells of C3H or BALB / C mouse
Add 10 ⁵ / well, add 3 in CO ₂ incubator
The cells were cultured at 7 ° C for 3 days. 4 hours before the end of the culture, 1 μCi
/ Well of ³ H-thymidine was added, and after the culture was completed, the amount of tritium-labeled thymidine incorporated in the cells collected on the filter was measured with a β-counter to mix mesenteric lymph nodes of aged mice. The relative stimulation index of the lymphocyte culture response was calculated. Then, the mixed lymphocyte culture reaction in a 7-9 week old young mouse different from the aging mouse was measured in the same manner as described above, and the relative stimulation index was calculated. Based on the relative irritation index of aged mice, changes in the relative irritation index of aged mice fed with immunized milk or control milk were compared. Table 5 shows the results. [Table 5] Next, for each of the cases where C3H and BALB / C are used as indices, the numerical value of the relative stimulation index of the young mouse (D) to the numerical value of the relative stimulation index of the aged mouse (E)
(F) was compared in the groups receiving the immunized milk or control milk, and how much the mixed lymphocyte culture reaction of the mesenteric lymph nodes suppressed the age-related decrease in the group administered with the immunized milk. It was investigated. Table 6 shows the results. [Table 6] From these results, it can be seen that the degree of the decrease in the mixed lymphocyte reaction of the mesenteric lymph nodes which decreased with aging was greater in the group receiving the immunized milk than in the group receiving the control milk. It was confirmed that it was significantly suppressed. Experiment 3: Mitogen responsiveness of mesenteric lymph nodes Method: 8-week-old C57BL / 6CrSIC
Fourteen female mice were divided into two groups, and each group (7 mice) was mixed with 48.9 g of powder of immunized milk or control milk and mixed feed 1
1.1 g, 3 g / animal per day
After ingestion for 9-71 weeks, the mesenteric lymph nodes that were sacrificed, the abdomen was opened and removed, and 10 cc of RPMI 164 were removed.
Place in a Petri dish containing medium 0. Crush with two slide glasses in a Petri dish. The released cells were strained with a gauze and collected to obtain lymphocytes in mesenteric lymph nodes. Lymphocytes contained in the mesenteric lymph nodes of the aged mice were used, each at 5 × 10 ⁵ / well. 0.4 μg / well of concanavalin A (ConA), 0.02% (V / V) / well of phytohemagglutinin (PHA) or 10 μg / well of lipopolysaccharide are added, and the mixture is added at 37 ° C. in a CO ₂ incubator at 3 ° C. Cultured for days. Four hours before the end of the culture, 1 μCi / well of ³ H-thymidine was added. After the end of the culture, the amount of tritium-labeled thymidine incorporated in the cells collected on the filter was measured with a β-counter, and aging was performed. The relative stimulus index of mitogen responsiveness of mouse mesenteric lymph nodes was calculated. The mitogen responsiveness of mesenteric lymph nodes in a young mouse different from the aging mouse was measured in the same manner as described above, and the relative stimulation index was calculated. The change of the relative stimulation index of the aged mice fed with the immunized milk or the control milk was compared based on the relative stimulation index. Table 7 shows the results. [Table 7] Next, when ConA, PHA or LPS was used as an index, the value obtained by subtracting the value (H) of the relative stimulation index of the aged mouse from the value (G) of the relative stimulation index of the young mouse for each. By comparing (I) in the group to which the immunized milk or the control milk was administered, it was examined how much the group to which the immunized milk suppressed the age-related decrease in mitogen responsiveness of the mesenteric lymph nodes. . Table 8 shows the results. [Table 8] Note that ConA (concanavalin A) and P
HA (phytohemagglutinin) is a plant-derived lectin, which is a type of mitogen that stimulates and activates T cells to cause DNA synthesis and cell growth. Since lymphocytes of the mesenteric lymph node contain B cells in addition to T cells, the response to LPS (lipopolysaccharide) was also measured as an index of the immune response ability of B cells. In the group to which the immunized milk was administered, the degree of the decrease in mitogenic response of the mesenteric lymph node which decreased with age was compared between the group to which the immunized milk was administered and the group to which the control milk was administered. It was confirmed that the group to which the immunized milk was administered was significantly suppressed. [Effect] The immune function plays a role of defense against infection and a mechanism of immune monitoring in the living body. When the immune function is reduced by aging, it becomes a trigger for the occurrence of disease. The composition of the present invention for suppressing age-related immune decline is said to be easily developed with age, such as respiratory infections such as pneumonia, skin diseases such as shingles, urinary tract infection, and biliary tract. It has the effect of preventing or treating the onset of diseases called so-called geriatric diseases, such as infectious diseases, postoperative complications when an elderly person undergoes a surgical operation, and the onset of suppuration of wounds and colds. Further, the composition of the present invention can be orally administered and is highly safe, so that it can be easily taken every day and has excellent practicality.

──────────────────────────────────────────────────続 き Continued on the front page (73) Patent holder 000165376 Kanematsu Corporation 119, Ito-cho, Chuo-ku, Kobe City, Hyogo Prefecture (72) Inventor Atsushi Ishida 2-6-5 Minami, Honjo City, Saitama Prefecture (72) Inventor Shinji Murosaki 1-24-27-102 Meinohama, Nishi-ku, Fukuoka City, Fukuoka Prefecture (56) References JP-A-54-113425 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) A61K 39 / 40 A61P 37/04 BIOTECHABS (STN) CA (STN) MEDLINE (STN)

Claims (1)

(57) [Claims] A composition for suppressing immunological decline comprising as an active ingredient milk expressed from dairy cows sensitized with one or more bacterial antigens selected from the group of microorganisms shown in the following table (Excluding prophylactic treatment of rheumatoid arthritis due to mixed bacterial infection of the gastrointestinal tract).