JPH05252900A - Immunopotentiative composition - Google Patents

Abstract

PURPOSE:To provide the subject composition for medicines or foods/beverages (e.g. preparation of infant rearing powdered milk) useful for the prevention and treatment of various diseases resulting from various kinds of antigenic substances or infectious microorganisms, comprising a cytoplasmic fraction or a protein complex and peptide complex each containing the cytoplasm fraction, obtained from lactobacillus microbes. CONSTITUTION:At least one kind of lactic acid bacteria selected from those belonging to Lactobacillus, Bifidobacterium, Pediococcus, Streptococcus, and Leuconostoc is cultured, and the lactic bacteria microbes in the resulting cultured product and/or culture solution are ground. Then, the cytoplasmic fraction of the microbes and/or a product containing the fraction in the resulting supernatant are diluted or concentrated, fractionated, purified and/or dried, thus obtaining the objective composition having the above-mentioned advantages.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an immunostimulatory composition, and more specifically to an in vivo immune system containing a cytoplasmic fraction of lactic acid bacterium and / or its content. The present invention relates to a composition having an excellent ability to activate.

The composition according to the present invention is used as an immunostimulant in a medicinal type, as well as beverages, foods for immunostimulation,
It is used in nutritional foods, functional foods, foods for specified health uses, food and drink types such as drinks, and has the ability to protect the organism against antigenic substances such as food allergens and indigenous bacterial flora, or infectious microorganisms such as viruses and pathogenic bacteria. It can be widely used for application to a living body.

[0003]

2. Description of the Related Art Various reports have recently been made on the biological immunostimulatory action of lactic acid bacteria, and the mechanism of immunostimulatory action and the difference in physiological activity depending on the bacterial species are gradually being clarified (Tetsuji Hirota, New Food. Industry, Vo
l. 32, No. 10, P9 (1990)). But,
Most of these reports are still only in vitro and in vivo analysis of the effect of lactic acid bacteria on the living body, and few studies have examined the active ingredient in the lactic acid bacteria. Furthermore, the cell wall fraction of bacterial cells and the polysaccharides produced by the bacterial cells are also targeted in these few studies (CI Park).
Et al., Milchwissenschaft, 46 , 87.
(1991); Yasui et al. Dairy S
ci. , 74 , 1187 (1991); Nagao
ka et al. Biochem. , 108 , 568 (19
90); Haruki Kitazawa et al., Research on dairy chemistry and food, 40 , A
-261, (1991)). As described above, the cytoplasmic fraction of lactic acid bacterium has not been examined, let alone the fact that the cytoplasmic fraction contains an immunostimulatory component, and that component is actually used for immunostimulation. The current situation is that no knowledge has been obtained.

On the other hand, breast milk contains a large number of protective substances against infection such as secretory IgA, lactoferrin, lysozyme and complement, which is said to be the best nutrition source for newborn infants with an intestinal immune system. It's one of the reasons. In infant formula powder used as a substitute when breast milk becomes insufficient or cannot be fed, substances such as lactoferrin have recently been added for the purpose of imparting a biological defense function.

Lactic acid bacteria are one of the substances that have been tried to be added from such a background, and are known as Bakhtyya in Russia.
Rova et al. showed that the amount of IgA antibody and lysozyme content in the feces of the newborn fed the modified milk powder supplemented with bifidobacteria was more than double that of the newborn fed the modified milk powder without the addition of bacterial cells. (Izvestiya Vysshikh Ucheb
nykh Zavedenii, Pishchevey
a Tekhnologya No. 1, P35 (1
988)). However, the bifidobacteria used by Bakhtyyarova et al. For the formula are only described as thermostable bifidobacteria (that is, viable bacteria of bifidobacteria), and the active ingredients in the bacterial cells as well as the strains used by bifidobacteria are described. Is not disclosed at all. On the other hand, although examples of use of milk containing Bifidobacterium have been reported in Japan, only the effects on macrophages of non-specific phagocytic cells and erythropoietic ability have been investigated. On the intestinal immune system and the active ingredients in the cells are completely unknown (Sekine et al., Therapeutics, Vo
l. 14, No. 5, P691 (1985)).

[0006]

As described above, only the cell wall or secreted polysaccharide has been emphasized as the active ingredient for the physiological action of lactic acid bacteria, and the cytoplasmic fraction generated in the process of fractionating them is particularly important. It didn't get the attention. However, the cytoplasm occupies a larger proportion than the cell wall when viewed from the whole cell body, and its effective utilization is a problem.

[0007] In practice, among the above-mentioned attempts to endow a newborn with a bioprotective ability by applying a lactic acid bacterium represented by Bifidobacterium to infant milk powder, It remains unclear whether it acts on the intestinal immune system. With this, there is a limit to the further utilization of lactic acid bacteria for infant milk powder and others.

[0008]

Means for Solving the Problems The present invention has been made in order to utilize lactic acid bacteria more effectively. The present inventors have fractionated cells of various lactic acid bacteria into a cell wall and a cytoplasm, and As a result of intensive studies on the effect of the fraction on immunocompetent cells, it was found that the cytoplasmic fraction exhibits an immunostimulatory action equivalent to or higher than that of the cell wall fraction, and the present invention has been completed.

[0009] That is, the present inventors have earnestly studied the active fraction in a lactic acid bacterium that activates Peyer's patch lymphocytes, which are the key to the intestinal immune system, when the lactic acid bacterium is orally administered to a living body. As a result, it was found that the cytoplasmic fraction of the bacterial cells showed a particularly high activity, and at the same time, in the intestinal tract (small intestinal contents and small intestinal wall) of the living body to which the bacterial cells were administered,
It was found that a significantly higher IgA antibody was produced as compared with the case where the cells were not administered, and the present invention was completed.

The present invention was the first to discover the above-mentioned usefulness for immunostimulation in the cytoplasmic fraction of lactic acid bacterium, and was completed as a result of further research based on this new finding. An immunostimulatory composition containing the cytoplasmic fraction as an active ingredient is the basic technical idea.

To obtain the cytoplasmic fraction of the lactic acid bacterium of the present invention, the cell wall-degrading enzyme is allowed to act on the microbial cell, or the microbial cell is mechanically treated with a French press or an ultrasonic crushing device, and after centrifugation. The supernatant may be collected. In this case, the lactic acid bacterium before fractionating the cytoplasmic fraction may be live or dead.
Furthermore, proteins or peptides in the cytoplasmic fraction by trichloroacetic acid treatment or salting-out treatment, or compounds containing them as one of the constituents (glycoprotein, lipoprotein, etc.)
Is more desirable if it can be purified. In the present invention, the cytoplasmic fraction includes all the products in each step from crushing of the lactic acid bacterium cells to fractionation and purification thereof, and the cytoplasmic fraction-containing material is also a constituent component of the present composition. It can be used as one of.

In the former, the lactic acid bacterium is disrupted and the supernatant obtained by centrifugation, the supernatant obtained by further centrifugation, and the lyophilized cytoplasmic fraction in a narrow sense, Fractionation, purification, treatment with trichloroacetic acid, salting out, etc. to obtain a protein complex (protein, lipoprotein,
Specific examples of cytoplasmic fractions include glycoproteins) and peptide complexes (peptides, glycopeptides, etc.). In addition, the cytosolic fraction in a narrow sense and in a broad sense may be diluted, concentrated and dried to be used freely.

In the latter, in addition to the above-mentioned substances containing the cytoplasmic fraction, all of them can be used as the cytoplasmic fraction-containing substance, and as the cytoplasmic fraction-containing substance, lactic acid bacterium and its treated product can also be used. As the treated product of lactic acid bacterium, a microbial cell-containing substance (lactic acid bacterium culture containing lactic acid bacterium, medium and culture supernatant; same culture solution obtained by removing medium components from the same culture; And the same supernatant obtained by removing the medium components), and crushed cells obtained by mechanically, chemically, and / or (micro) biologically crushing the cell wall can be widely used. .. Further, a substance containing bacterial cells, a precipitate fraction of crushed bacterial cells, dilution, concentration, and / or dried product can also be used.
As long as there is no problem with dispersibility, etc., the cytosolic fraction-containing material may be used for foods, pharmaceuticals, etc. without isolating and using the above-mentioned cytosolic fraction, and rather it may be used during oral administration. In many cases, the above-mentioned immunostimulant can be prevented from being decomposed by digestive enzymes, and the immunostimulatory component can act on the intestinal immune system in a highly active state in many cases. Both live and dead lactic acid bacteria can be used.

In the present invention, the lactic acid bacterium is not particularly limited as long as it is a microorganism belonging to lactic acid bacterium, and any microorganism can be used, and commercially available lactic acid bacterium and lactic acid bacterium used in foods and drinks or pharmaceuticals using lactic acid bacterium are also appropriate. Can be used.
Non-limiting examples of lactic acid bacteria that can be used in the present invention include:
These include: Lactobacillus (Lacto)
bacillus acidophilus ATCC
11506, L.I. lactis IFO 12522
Etc.); Bifidobacterium (Bifidobacterium)
erium longum ATCC 15708
Etc.); Pediococcus (Pediococcus)
cerevisiae ATCC 8042); Streptococcus fa
ecumium ATCC 8043); Leuconostoc mesenteroid
es IFO 3426 etc.) Others.

The immunostimulatory composition according to the present invention comprises one or more kinds of the above-mentioned cytoplasmic fraction and / or its content as an active ingredient, and is used as a food or drink or a medicine. When these active ingredients are used as foods, they can be added as they are, or can be used in combination with other foods or food ingredients in accordance with ordinary methods. When used as a medicine, it can be administered orally or parenterally. In the case of oral administration, for example, tablets, granules, powders, capsules and powders can be prepared according to a conventional method, and in the case of parenteral administration, for example, injection preparations, drops, suppositories, etc. It can be used as an agent or the like.

The active ingredient according to the invention has no or very low toxicity due to its natural origin and is extremely safe (LD ₅₀ > 3000 mg / kg subcutaneous,> 5000).
mg / kg orally: All are rats).

Hereinafter, the present invention will be described in more detail with reference to Examples.

[0018]

Example 1 Bifidobacterium longum ATCC
15708 was inoculated into an EG liquid medium ("World of Enterobacteriaceae" by Tomohashi Mitsuoka (Moubunsha 1980)), and anaerobic conditions were applied to 37
Incubated at 18 ° C for 18 hours. After the completion of the culture, the cells were collected by centrifugation, washed with cooled physiological saline three times, and suspended in distilled water. This suspension was treated with an ultrasonic crusher with an output of 100 W for 20 minutes, and the treated solution was centrifuged at 1,000 G to remove unbroken cells as a precipitate. Next, the supernatant was centrifuged at 30,000 G to obtain the supernatant (cytoplasmic fraction).
And pelleted (cell wall fraction) again. The supernatant was lyophilized to give a cytoplasmic fraction, and the precipitate was trypsin and D
After treatment with NA and RNA degrading enzyme, centrifugation and lyophilization were performed to obtain a cell wall fraction.

The mitogenic activity of both fractions thus obtained was measured as follows. C3
H / HeJ mouse splenocytes were collected, washed, and then suspended in RPMI1640 medium containing 1% of syngeneic mouse serum.
These cells were used in a 96-well flat bottom plate in the presence of the above cytoplasm and cell wall fractions (1 to 100 μg / wel).
l) Each was cultured in RPMI1640 medium containing 1% syngeneic mouse serum under 5% CO ₂ / Air conditions at 37 ° C. for 72 hours (5 × 10 ⁵ cells / well). Next 1.0 μC
i ³ H-thymidine was added, and 20 hours later, ³ H-thymidine uptake into cells was measured by a scintillation counter. As a control, ³ H-thymidine incorporation counts (c) of cells when no substance was added to the medium
pm), and the result (FIG. 1) was represented by a value (Δcpm) obtained by subtracting this from the well count number (cpm) of each sample.

According to the present invention, as shown in FIG. l
The mitogen activity equal to or higher than that of the cell wall was also observed in the ongum cytoplasm. Ovalbumin (OVA) is a negative control.

[0021]

Example 2 In the same manner as in Example 1, Bifidobacterium longum ATCC 15707, Lactobacillus acidophilus IFO 3953, Lactobacillus.
Delbricki Subspecies Ractis ATCC
The 12315 bacterial cells were cultured, ultrasonically disrupted and centrifuged to obtain the cytoplasmic fraction of each strain.

The cell growth activity of the cytoplasmic fractions of these strains against ovalbumin-sensitized T cells was measured as follows. For BALB / c mice, the method of Kurisaki et al. (Eur. J. Immunol. Vol. 16, P23) was used.
6, 1986) was subcutaneously immunized with ovalbumin (OVA) to obtain OVA-sensitized T cells. These cells were treated in the same manner as in Example 1 in the presence of the cytoplasmic fraction of each strain (2-1.
(00 μg / ml), and cell proliferation was examined by ³ H-thymidine incorporation. The results were represented by the well counts (stimulation coefficient: SI) of each sample when the well count (control count) in which no substance was added to the medium was 1.

According to the present invention, as shown in FIG. 2, the cytoplasm of each strain was β-, which was used as a negative control.
The cell proliferation activity was significantly higher than that of lactoglobulin (β-LG). In particular, two strains of Bifidobacterium longum were used as positive controls, and lipopolysaccharide (LP
The activity was almost comparable to that of S).

[0024]

Example 3 Bifidobacterium longum ATCC
15708 to produce a 2.0 × 10 ¹¹ cells / g containing for bacteria powder containing 5% to whey protein (WPI) chow, were bred mice (BALB / c, 5W, female) Eight ( The composition of WPI solid feed is based on AIN-76). Two weeks later, the Peyer's patch was aseptically removed from the small intestine of the mouse, gently teethed with a slide glass, and singled.
e cell. These cells were cultivated using a 96-well flat bottom plate in the presence of cytoplasmic and cell wall fractions of Bifidobacterium longum ATCC 15708 (1-100 μg / ml) prepared in Example 1 at 37 ° C. for 48 hours (others). The conditions are the same as in Example 1.) Then 1.0
μCi ³ H-thymidine was added, and 20 hours later, ³ H-thymidine uptake into cells was measured by a scintillation counter. The results are shown in Fig. 3. The display of results was in the same format as in FIG.

According to the present invention, as shown in FIG. 3, the cytoplasmic fraction of the bacterial cells had a strong cell proliferating activity on Peyer's patch lymphocytes. On the other hand, the cell wall fraction of the bacterial cells did not show cell proliferating activity, which was almost the same response as the negative control ovalbumin (Δcpm). From this, it was revealed that when the bacterial cells were orally administered, their cytoplasm strongly stimulated the intestinal immune system.

[0026]

Example 4 In the same manner as in Example 3, Peyer's patch lymphocytes were obtained from BALB / c mice to which Bifidobacterium longum ATCC 15708 was orally administered for 2 weeks. These cells were transformed into Bifidobacterium longum A
TCC 15708 cytosolic tryptic digest (0,
The cells were cultured under the same conditions as in Example 3 in the presence of (1, 5, 24 hours degradation), and cell proliferation was examined by ³ H-thymidine incorporation. The cytosolic trypsin degradation product is trypsin (S) in a buffer solution of pH 8.5 for a predetermined time (0 to 24 hours).
igma) was allowed to act at an E / S ratio of 1: 100, trypsin was inactivated by heating, desalted, and lyophilized. FIG. 4 shows the change in activity associated with trypsin treatment of cytoplasm. The display of results was in the same format as in FIG.

According to the present invention, as shown in FIG. 4, cytoplasmic cell proliferating activity decreased with extension of trypsin treatment time. From this, it was clarified that proteins and peptides in the cytoplasm or various complexes thereof (glycoprotein, lipoprotein, etc.) are the main components of the activity of this immunostimulant.

[0028]

[Example 5] Chymotrypsin (Sigma) was used in place of the trypsin used in Example 4, and changes in activity due to chymotrypsin treatment of cytoplasm were examined. However, the enzyme treatment time was 0.5, 24 hours. The result is shown in Figure 5.
It was shown to.

According to the present invention, as shown in FIG. 5, the cytoplasmic cell proliferating activity decreased with the increase of the chymotrypsin treatment time. From this, it was revealed that proteins and peptides in the cytoplasm or various complexes thereof (glycoprotein, lipoprotein, etc.) were the main components of the activity of this immunostimulant, as in Example 4.

[0030]

Example 6 Bifidobacterium longum ATCC 15708 2.0 × 10 ¹¹ /
The culture solution grown to a volume of ml was centrifuged and concentrated at 4000 G, the same amount of deionized water as that before centrifugation was added to the obtained precipitate fraction to uniformly disperse the precipitate, and then HTST sterilization was performed. Then, the mixture was centrifuged again, and the precipitate fraction was collected to obtain freeze-dried bacterial cell powder. B. in powder The number of longum bacterial cells was about 8.0 × 10 ¹¹ cells / g. 0.1% of this powder was added to commercially available infant formula (Meiji Dairy Co., Ltd.).
Compounded.

[0031]

Example 7 Lactobacillus acidophilus IFO 3953 1.5 × 10 ¹¹ cells / ml by high-concentration culture method
The culture solution that had been grown up to (3) was concentrated by centrifugation at 3000 G, and the obtained precipitate fraction was sterilized by HTST and then lyophilized. L. in this freeze-dried cell powder. The number of acidophilus cells was equivalent to about 4.0 × 10 ¹¹ cells / g. 0.05% of this powder was added to commercial infant formula (Meiji Dairy Co., Ltd.).
Compounded.

In the production of the immunostimulatory infant milk powder according to the present invention, the lactic acid bacterium-derived cytoplasm fraction can be used in the above-mentioned amount as an example. 1.0 x 10 ⁴ or more per 1 g of milk powder,
Preferably, a cytoplasmic fraction derived from 1.0 × 10 ^{6 to} 1.0 × 10 ¹² cells of lactic acid bacteria may be used. However, since the cytoplasmic fraction is derived from a natural product, it is safe and therefore, there is no problem even if it is used in a larger amount than the above range. May be used in small amounts. Also, when preparing foods and drinks other than infant formula, the amount of cytoplasmic fraction to be used may be determined with reference to the above range.

[0033]

Example 8 Lactobacillus acidophilus IFO
A whey protein (WPI) solid feed containing 5 × 10 ¹⁰ heat-killed bacterial cells of 3953 / g of feed was prepared, and L. cerevisiae against Peyer's patch lymphocytes of the cells-administered mouse was prepared in the same manner as in Example 3. The cell growth activity of the cell wall and cytoplasmic fraction of acidophilus IFO 3953 was examined 2 weeks after the start of oral administration of bacterial cells.

The result is shown in FIG. According to the present invention,
L. The cytoplasmic fraction of acidophilus IFO3953 showed a strong proliferative activity on Peyer's patch lymphocytes, while the cell wall fraction was negative control.
It showed a slightly stronger activity than the OVA used as.

[0035]

Example 9: B. longgumAT
Seven mice (BALB / c, 5W, female) were bred on whey protein (WPI) solid feed containing CC 15708. Four weeks later, the small intestine was removed from the mouse, and the method of Tsuki et al. (Mitsuoka Tomohashi, “Research methodology of intestinal flora”,
P. 157 (Society Press Center 1989)) to obtain small intestine contents and small intestinal wall extract. The IgA antibody titer in these samples was measured by an ELISA method using an anti-mouse IgA monoclonal antibody (Kanto Kagaku). As a control, B. The IgA antibody titers in the small intestinal contents and the small intestinal wall extract obtained from the mice that had been raised in the same manner as above with the WPI solid diet having the same composition but containing no longum ATCC 15708 were also examined.

The result is shown in FIG. According to the present invention, it was revealed that activation of Peyer's patch lymphocytes by the cytoplasmic fraction of lactic acid bacterial cells resulted in a higher IgA antibody titer in the intestinal tract than in the control.

[0037]

Example 10 Mouse (BALB / c, 5W, female) 21
The 7 animals were divided into 3 groups, each of which was used in Examples 3 and 8. longum fungus-containing WPI solid feed and L. WPI solid feed containing the acidophilus cells was given to each of the two groups and bred. The remaining one group was fed with WPI solid feed having the same composition and not containing these cells. One year later, the small intestine part was taken out from each group of mice, and a small intestine wall extract was obtained in the same manner as in Example 9. The anti-β-lactoglobulin IgA antibody titer and anti-LPS IgA antibody titer in these samples were measured by the ELISA method.

The result is shown in FIG. According to the present invention,
As a result of activation of Peyer's patch lymphocytes by the cytoplasmic fraction of lactic acid bacteria, β, which is said to be a food allergen in WPI
-It was revealed that IgA antibody titers in the intestinal tract against lactoglobulin and LPS of Gram-negative bacteria such as Escherichia coli are higher than those of the control.

[0039]

[Example 11] Vitamin C 20 g, granulated sugar 50
g, 50 g of the cytoplasmic fraction obtained in Example 1 was added to 30 g of an equivalent mixture of corn starch and lactose, and they were mixed sufficiently.
The mixture was divided into 100 equal parts and packed in a bag to produce 100 bags of 1.5 g of stick-shaped nutrition-stimulating nutritional and health food.

[0040]

Example 12 The cytoplasmic fraction obtained by using Pediococcus cerevisiae ATCC8042 instead of Bifidobacterium as the microorganism in Example 1 is used.
Further, the same procedure as in Example 11 was repeated except that 20 g of an equivalent mixture of vitamin C and citric acid was used instead of 20 g of vitamin C, and the mixture was sufficiently dried and then an immunostimulated nutritional health food was produced.

[0041]

Example 13 An immunostimulant was produced by the following formulation.
(1) Streptococcus faecium ATCC 804 in place of Bifidobacterium in Example 1
50 g of the freeze-dried product of the ammonium sulfate salted-out product of the cytoplasmic fraction obtained using 3., (2) 90 g of lactose, (3) 29 g of corn starch, (4) 1 g of magnesium stearate.

First, (1), (2), and (3) (however, 17 g) were mixed and granulated with the paste prepared from (3) (however, 7 g). To the obtained granules, (3) (however, 5 g) and (4) were added and mixed well, and this mixture was compressed by a compression tableting machine to obtain 10 parts of cytoplasmic fraction per tablet.
1000 tablets containing mg were produced.

The dose varies depending on the patient's symptoms and age, but is 0.1-1500 mg / kg / day, 1 day a day.
~ 4 doses. Since the cytoplasmic fraction used in the present invention is originally derived from foods, there is almost no problem in safety as described above, and therefore, administration in excess of the above dose is acceptable. When it is used as a health maintenance, promotion, health care, nutritional supplement, etc., a dose smaller than the above dose may be taken for a long period of time. Also,
As described above, the tablet according to the present invention can be administered by a method other than oral administration, but in the case of intravenous administration and intramuscular administration, it is 0.01 to 1200 mg / kg / day.

[0044]

Example 14 The following formulation was prepared. (1) 10 g of the cytoplasmic fraction obtained in Example 1, (2) 8 g of sodium chloride,
(3) Chlorobutanol 4 g, (4) Sodium hydrogencarbonate 1 g.

All the components were dissolved in 1000 ml of distilled water and dispensed into two 500 ml infusion bottles to prepare an immunostimulatory infusion solution.

[0046]

INDUSTRIAL APPLICABILITY The immunostimulatory composition of the present invention is effective against the cytoplasmic fraction, unlike the cell wall fraction or the polysaccharide secreted by the bacterium, which has been conventionally studied as the active entity of the immunostimulatory action of lactic acid bacteria. As an ingredient. Therefore, there is a great possibility that the mechanism of action on various immunocompetent cells is different from the above-mentioned substances that have been studied so far, and it can be widely applied to foods and pharmaceuticals as a highly safe immunostimulant derived from lactic acid bacteria.

In addition, according to the present invention, a cytoplasmic fraction of lactic acid bacterium cells which enhances the biological defense ability against an antigenic substance such as a food allergen or an indigenous bacterial flora, or an infectious microorganism such as a virus or a pathogen is used as an active ingredient By directly using it as a food or drink, or by adding it to a food or drink, it is possible to supply a food or drink useful for the prevention or treatment of various diseases caused by these antigenic substances or infectious microorganisms. For example, by blending the cytoplasmic fraction with baby milk powder, it is possible to produce baby milk powder that is extremely effective in preventing various diseases caused by these antigenic substances or infectious microorganisms such as artificial nutrition.

[Brief description of drawings]

FIG. 1B. It is the figure which compared the mitogenic activity of the cytoplasm and cell wall fraction of a Longum fungus body.

FIG. 2 is a diagram showing promotion of cell proliferation of ovalbumin-sensitized T cells by cytoplasmic fractions of various lactic acid bacteria.

FIG. 3B. It is a figure showing promotion of cell proliferation at the time of oral administration of longum bacterial cells (Example 3).

FIG. 4B. It is a figure showing the influence of trypsin processing of a cytoplasmic fraction on promotion of cell growth at the time of oral administration of longum bacterial cells.

FIG. 5 B. It is a figure showing the influence of chymotrypsin treatment of a cytoplasmic fraction on promotion of cell growth at the time of oral administration of longum bacterial cells.

FIG. It is a figure showing promotion of cell growth at the time of oral administration of acidophilus bacterial cells (Example 8).

FIG. 7: B. Intestinal Ig by oral administration of longum bacterial cells
It is a figure showing induction of A production.

FIG. 8: Intestinal tract I against antigenic substance by oral administration of bacterial cells
It is the figure which showed the induction of gA production.

[Explanation of symbols]

B: Bifidobacterium longum bacterial cell administration group L: Lactobacillus acidophil
Us bacterial cell administration group C: control group

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Office reference number FI technical display location C12N 1/20 A 7236-4B (72) Inventor Yu Kuwata 1-21 Sakaemachi, Higashimurayama, Tokyo 3 Meiji Dairy Products Co., Ltd. Central Research Institute (72) Inventor Ryorou Yamamoto 1-21-3 Sakaemachi, Higashimurayama-shi, Tokyo Meiji Dairy Co., Ltd. Central Research Institute

Claims (14)

[Claims] 1. An immunostimulatory composition comprising a cytoplasmic fraction of lactic acid bacterium and / or a substance containing a cytoplasmic fraction. 2. The composition according to claim 1, wherein the active ingredient of the cytoplasmic fraction is a protein complex and / or a peptide complex. 3. The composition according to claim 2, wherein the protein complex is a protein, a lipoprotein and / or a glycoprotein. 4. The composition according to claim 2, wherein the peptide complex is a peptide and / or a glycopeptide. 5. The cytoplasmic fraction comprises a lactic acid bacterium culture and / or a supernatant obtained after disrupting the lactic acid bacterium in the culture medium.
The composition according to any one of 1. 6. The cytoplasmic fraction is further diluted, concentrated, fractionated, purified and / or dried to obtain a cytoplasmic fraction.
The composition according to claim 5. 7. The substance containing a cytoplasmic fraction is a substance containing the cytoplasmic fraction according to any one of claims 1 to 5, a lactic acid bacterium, and / or a treated product of the bacterium. An immunostimulatory composition comprising: 8. The treated product of lactic acid bacteria is a substance containing a microbial cell, a crushed bacterial cell, a precipitation fraction of the sol, dilution of the ora,
The composition according to claim 7, which is a concentrated and / or dried product. 9. The lactic acid bacterium-containing substance is a lactic acid bacterium culture product,
The composition according to claim 8, which is the same culture solution and / or the same supernatant. 10. The lactic acid bacterium is Lactobacillus (Lact).
obacillus), Bifidobacterium (Bif
idobacterium), Pediococcus (Pe)
diococcus), Streptococcus (Stre)
ptococcus, Leuconostoc (Leuco)
The immunostimulatory composition according to any one of claims 1 to 9, which is one kind or two or more kinds belonging to each genus Nostoc). 11. The immunostimulatory composition is an immunostimulant and / or an immunostimulatory food or drink, which is characterized in that
The immunostimulatory composition according to claim 10. 12. The composition according to claim 11, wherein the immunostimulating food or drink is a dairy product. 13. The composition according to claim 12, wherein the dairy product is baby milk and / or baby milk powder. 14. 1.0 × 10 ⁴ or more, preferably 1.0 × 10 ^{6 to} 1.0 × 10 ¹² cytoplasmic fractions derived from lactic acid bacterium cells are contained per 1 g of milk powder. 14. The composition according to claim 13, which comprises: