WO2006006273A1 - Dendritic cell activator - Google Patents

Abstract

[PROBLEMS] To provide a method of safely and efficiently activating dendritic cells and provide a dendritic cell activator capable of presenting means for the method. [MEANS FOR SOLVING PROBLEMS] There is provided a dendritic cell activator comprising an extract from shiitake mushroom mycelia. Further, there are provided, comprising the dendritic cell activator, a pharmaceutical composition, food composition, food, beverage, etc. Still further, there is provided a method of inducing mature dendritic cells, comprising treating immature dendritic cells with the dendritic cell activator. Yet still further, there are providedmature dendritic cells obtained by the above method and a pharmaceutical composition comprising the mature dendritic cells.

Description

 Specification

 Dendritic cell activator

 Technical field

 [0001] The present application relates to a rod-shaped cell activator comprising a shiitake mycelium extract or a fraction thereof, particularly a rod-shaped cell maturation promoter. Furthermore, the present invention relates to a composition for oral intake, a pharmaceutical composition, a food composition and the like containing the rod-shaped cell activator.

 Background art

 [0002] Spider cells are a type of immune cell having a morphological feature of having dendritic processes, and are distributed in various tissues (for example, skin and mucous membranes) to serve as immune surveillance cells. Yes. In particular, rod cells are cells that have the function of transporting antigen that has entered the body to lymphoid tissues and stimulating T cells to induce an immune response, and play a central role in the immune response together with lymphocytes.

 [0003] Among the rod cells distributed in various in vivo tissues, the rod cells present in the peripheral tissue are called immature rod cells because of their low T cell activity. In contrast, rod cells present in lymphatic organs are called mature rod cells because they have a strong ability to activate T cells. These functional differences are due to increased expression of cell adhesion and costimulatory molecules (eg, CD40, CD54, CD58, CD80 and CD86) as well as cytokines in addition to MHC (major histocompatibility complex) molecules This is due to changes in production capacity such as chemokines. As the maturation progresses, the rod-shaped cells change from a strongly adherent structure to a highly motile structure due to reorganization of the cytoskeleton, and begin to migrate to the lymph nodes. In the T region of the regional lymphoid organs, the rod-like cells promote the activity of T cells by producing chemokines (Non-patent Document 1: Cell Engineering, No. 19, No. 9, 2000, No. 1). 1311-1317).

[0004] In vivo, maturation of rod-shaped cells is caused by TNF a produced by LPS (lipopolysaccharide), DNA, double-stranded RNA, or tissue cell force derived from induction of inflammatory response from infected bacteria or viruses. (Tumor necrosis factor a) and IL-1 (interleukin 1), as well as CD40 ligand (CD40L) expressed in active platelets, basophils, mast cells, etc. Reference 2: Banchereau J. et al., Nature, 392, 199 8 years, pp. 245-252).

[0005] In addition, in an in vitro experiment, an attempt has been made to search for a substance having a rod-like cell activation action. For example, a preparation obtained by treating Krestin (hereinafter also referred to as “rPSKj”) derived from the extract of Mycelia agaricus mycelium and an attenuated natural mutant of group A hemolytic streptococci (Su strain) with belin. There has been reported to have the effect of promoting the maturation of榭状cells (non-Patent Document _3: Bioth _e mpy, 15 Certificates, No. 3, 2001, 385- 388 pp; non-Patent Document 4: Cancer Immunology and Immunotherapy, 52nd, 2003, 207-214). In addition, it has been reported that the cell wall skeleton of certain gram-positive bacteria promotes maturation of rod-shaped cells (Patent Document 1: International Patent Publication 01Z048 154 pamphlet). Furthermore, it has been reported that a specific low molecular weight compound obtained by chemical synthesis has a rod-like cell activating action (Patent Document 2: Japanese Patent Application Laid-Open No. 2004-210768).

On the other hand, shiitake mushrooms, which are representative edible mushrooms in Japan and China, have been ingested as a food for many years in Japan, and have been cultivated artificially for 300 years. It is considered to be excellent in safety when administered orally, especially when administered orally for a long period of time. There have been several reports on the pharmacological action of shiitake mushrooms (for example, Patent Document 3: Japanese Patent Laid-Open No. 62-270532; Patent Document 4: Japanese Patent Laid-Open No. 2-237934; Patent Document 5: JP-A-2-134325; Patent Document 6: JP-A-12-12538, etc.). In particular, lentinan containing shiitake mushroom-derived j8-glucan as an active ingredient is commercially available as an anticancer agent, and is thought to induce and activate macrophages, killer T cells, natural killer cells, etc. (Non-patent Document 3 _: Biothempy, No. 15, No. 3, 2001, pp. 385-388). Therefore, there has been no report on shiitake extract that has any effect on rod cells.

[0006] In addition, for the purpose of clinical application of the powerful antigen-presenting ability of rod cells, research on anti-cancer therapy and anti-infection therapy using rod cells has been advanced! (Non-Patent Document 5: Nikkei Science, February 2003, pp. 84-91; Non-Patent Document 6: Contemporary Medicine, August 2003, pp. 86-92). In such therapy, by some means It is necessary to activate the rod-shaped cells, and in particular, a safe and efficient rod-shaped cell activation agent is required.

 Patent Document 1: International Patent Publication 01Z048154 Pamphlet

 Patent Document 2: JP 2004-210768 A

 Patent Document 3: Japanese Patent Laid-Open No. 62-270532

 Patent Document 4: Japanese Patent Laid-Open No. 2-237934

 Patent Document 5: JP-A-2-134325

 Patent Document 6: Japanese Patent Laid-Open No. 2003-012538

 Non-Patent Document 1: Cell Engineering, 19th, 9th, 2000, 1311-1317

 Non-Patent Document 2: Banchereau J et al., Nature, No. 392, 1998, pp. 245-252 Non-Patent Document 3: Biotherapy, No. 15, No. 3, 2001, pp. 385-388

 Non-patent document 4: Cancer Immunology and Immunotherapy, 52nd article, 2003

Pp. 207-214

 Non-Patent Document 5: Nikkei Science, February 2003, pp. 84-91

 Non-Patent Document 6: Contemporary Medicine, August 2003, pp. 86-92

 Disclosure of the invention

 Means for solving the problem

 [0007] The present inventor conducted extensive research to achieve the above-mentioned problems. As a result, the inventors discovered that an extract of shiitake mycelium has a rod-like cell activating action and completed the present invention. An object of the present invention is to provide an extract derived from shiitake mycelium having a rod-like cell activation action, and a rod-like cell activator, immunostimulant, composition for oral consumption, and pharmaceutical composition containing the extract. Products, food compositions, foods and beverages.

[0008] That is, according to one aspect of the present invention, a rod-like cell active agent containing shiitake mycelium extract is provided. Here, the term “activation of rod cells” means that some function of the rod cells is enhanced, or the amount of molecules such as proteins expressed in the rod cells is amplified. The intent of the term includes, for example, promoting maturation of rod cells, enhancing antigen uptake ability, enhancing antigen presentation ability, and enhancing τ lymphocyte stimulation ability. As a phenomenon observed in the rod-shaped cells with the activation, the adhesion molecule Z Increased expression of stimulatory molecules (eg, CD40, CD54, CD58, CD80, CD86), enhanced production capacity of site force-in (eg, interleukin-12, interferon gamma and interleukin-1), MHC For example, increase in the expression level of the molecule.

 [0009] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the above rod-shaped cell activator. The pharmaceutical composition is not particularly limited, and can be used, for example, for immunostimulation, treatment or prevention of cancer or malignant tumor, or treatment or prevention of infectious diseases. Here, the cancer and malignant tumor are not particularly limited, but for example, digestive organ cancer such as colon cancer and stomach cancer, myeloma, liver cancer, leukemia, melanoma, prostate cancer, breast cancer, uterine cancer, lung cancer, oral cavity Examples include cancer and brain tumors. In addition, although not particularly limited, infectious diseases include viral infections such as chronic hepatitis due to hepatitis C and hepatitis C virus, HIV, influenza, and the like.

 [0010] The mycelium of shiitake mushroom in the present invention is not particularly limited, and, for example, a mycelium in a state before the edible fruit body can be used. The mycelium may be produced by culturing or collected from nature. In the present invention, for example, a mycelium obtained by culturing shiitake mushrooms in a solid medium can be used. The shiitake mycelium extract used in the present invention can be prepared by a method known in the art, but an extract obtained by hot water extraction after pulverization of the mycelium can be used. Furthermore, for example, an extract obtained by crushing and decomposing a solid medium containing mycelium in the presence of water and an enzyme can be used. As a solvent used for the preparation of the extract, for example, water, ethanol, methanol, butanol, isopropanol, etc., preferably water can be used. Extraction should be performed with ultrasonic waves at a lower temperature (for example, 25-50 ° C, preferably 30-45 ° C) than the force that can be performed under heating of the solvent (for example, about 85-105 ° C). You can also do this.

[0011] Although the shiitake mycelium extract is not limited, for example, an extract obtained by the following method can be used. That is, inoculate shiitake mushrooms on a solid medium based on bagasse (sugar cane scum) and defatted rice bran, and then grow the mycelium to obtain a solid medium containing mycelium that has a weight of 30%. %, And water and cellulase, protease, or darcosidase power are selected in the solid medium. At least 70% by weight of bagasse fiber by adding one or more of the enzymes to be added while maintaining the solid medium at a temperature of 30-55 ° C, and crushing and grinding the solid medium in the presence of the enzyme. Is then passed through 12 mesh, then the enzyme is inactivated and sterilized by heating to a temperature of up to 90 ° C, and the resulting suspension is filtered to obtain a shiitake mycelium extract. obtain. The extract may be used as a shiitake mycelium extract, but it is convenient to store it as a powder by concentrating, lyophilizing, and using it in various forms at the time of use. The powder obtained by freeze-drying is a brown powder, is hygroscopic and has a unique taste and smell. The extract of the mycelium of shiitake mushroom thus obtained contains 15-50% carbohydrate, preferably 20-40% (w / w) by carbohydrate analysis by the phenol sulfate method, and 10-10 protein by lowry protein analysis. — 40%, preferably 13-30% (wZw), Folin—standard with gallic acid Contains 15% polyphenols, preferably 2.5-3.5% (wZw) by Denis method . Shiitake mycelium extract is not limited to this, including about 0.1% lipid, about 0.4% fiber, and about 20% ash.

[0012] An example of the constituent sugar composition (%) of the extract of shiitake mycelium was as follows, but this composition may vary depending on the culture conditions and the like: xylose: 15.2; arabinose: 8.2 Mannose: 8.4; glucose: 39.4; galactose: 5.4; amino sugar: 12.0; uronic acid: 11.3

[0013] The shiitake mycelium extract in the present invention may be a fraction obtained from the ability of shiitake mycelium extract. The fraction is obtained by a fractionation method usually used in the technical field to which the present invention belongs, and examples of the fractionation method include fractionation by extraction using an arbitrary solvent, gel filtration column chromatography, ion This includes fractionation using an exchange column and silica gel column chromatography. The fractionation method may be one type or a combination of multiple means. The solvent used in the fractionation method is not particularly limited, and for example, water, methanol, ethanol, n-propanol, isopropanol, acetic acid, acetone, and a mixture thereof can be used. The fraction can be used as a soluble fraction by concentrating, sticking, or solid, obtained by concentration and Z or lyophilization, if necessary.

[0014] As used herein, the term "spider cell activator" generally refers to spider cells. By acting, it means a drug that has the effect of enhancing the function of rod cells or amplifying the amount of substance expressed on or in rod cells, For example, effects such as promoting maturation of rod-shaped cells and increasing the ability to produce cytodynamic force (eg, interleukin-12) can be obtained.

[0015] The rod-shaped cell activator of the present invention can be used as an active ingredient of a pharmaceutical composition. The pharmaceutical composition can be used in various dosage forms such as tablets, capsules, powders, granules, pills, solutions, emulsions, suspensions, solutions, spirits, syrups for oral administration. And parenteral preparations such as subcutaneous injections, intravenous injections, intramuscular injections and intraperitoneal injections. However, it is not limited to these. These preparations can be produced by known methods usually used in the preparation process.

[0016] The pharmaceutical composition may contain various commonly used ingredients, such as one or more pharmaceutically acceptable excipients, disintegrants, diluents, lubricants, dressings, and the like. Flavoring agents, coloring agents, sweetening agents, flavoring agents, suspending agents, wetting agents, emulsifying agents, dispersing agents, adjuvants, preservatives, buffering agents, binders, stabilizers, coating agents and the like can be included. Further, the pharmaceutical composition of the present invention may be a sustained or sustained release dosage form.

[0017] The dosage of the pharmaceutical composition of the present invention can be appropriately selected depending on the administration route, the patient's body shape, age, physical condition, degree of disease, elapsed time after onset, etc. The pharmaceutical composition of the present invention The article can include a therapeutically effective amount and a Z or prophylactically effective amount of a rod cell activator. Shiitake mycelium extract was originally used as a food and has little concern for hyperplasia! / It is a relatively safe substance, so it can be administered at high concentrations as needed. is there. In the present invention, shiitake mycelium extract may be used at a dose of generally 100-5000 mgZ day Z adult, preferably 300-3000 mgZ day Z adult. The pharmaceutical composition may be administered in a single dose or multiple doses such as other rod-like cell activity agents, immunostimulants, anticancer agents, antitumor agents, anti-infective agents, etc. It can be used in combination with drugs.

[0018] According to still another aspect of the present invention, there is provided a method of inducing mature rod cells by treating immature rod cells with the above rod-shaped cell activator. Here, unfinished The mature rod-shaped cells are not particularly limited. For example, monocyte cells are cytosylated (for example, condylar sphere Z macrophage colony stimulating factor (hereinafter also referred to as “GM CFS”) and interleukin 4 (hereinafter referred to as “ It is also referred to as “IL 4”))). Here, the monocyte cell is not particularly limited. For example, a blood monocyte cell can be used, and a human blood monocyte cell is preferable.

 [0019] The change of immature rod cells into mature rod cells can be confirmed by methods known to those skilled in the art in the art. For example, costimulatory factors common to rod cells (for example, CD40, CD80, CD86) and an increase in the expression level of a protein specific to mature rod cells (eg, CD83) can be determined. The expression levels of these proteins can be observed by, for example, a flowmetry method.

 [0020] The rod-shaped cell activator of the present invention used when activating rod-shaped cells in vitro is, as shown in the table, in an amount of 10-1000 μgm, preferably 100-500 μgZmL per day. It can be added to the culture medium. Further, for example, the activity of rod-shaped cells can be induced by culturing for a period of 1 to 15 days, preferably 2 to 3 days.

 [0021] The method of the present invention may further include a step of mixing the rod-shaped cells and the antigenic substance in order to cause the obtained mature rod-shaped cells to present a specific antigen. Here, the antigenic substance is not particularly limited, but tumor-derived peptides or virus-derived peptides can be used, and for example, CEA-derived peptides, MAGE-derived peptides and the like can be used. Mixing of the rod-shaped cells and the antigenic substance may be performed before or after the activation of the rod-shaped cells or simultaneously with the activation of the rod-shaped cells, but is preferably performed simultaneously with the activation of the rod-shaped cells.

[0022] According to still another aspect of the present invention, a mature rod cell obtained by treating an immature rod cell with a rod cell activator, and a pharmaceutical composition comprising the mature rod cell Is provided. Here, the pharmaceutical composition is not particularly limited, but can be used for preventing or treating cancer, malignant tumors or infectious diseases. For example, digestive organ cancer such as colon cancer and gastric cancer, bone marrow Cancer, liver cancer, leukemia, melanoma, prostate cancer, breast cancer, eclampsia, lung cancer, oral cancer, brain tumor and other cancers or malignant tumors; or chronic hepatitis due to hepatitis B or C virus, HIV, influenza, etc. Can be applied to [0023] According to a further aspect of the present invention, there is provided an auxiliary activator for use in immunotherapy, comprising the above-described rod-shaped cell activator. Here, the auxiliary activator is a substance (adjuvant) that is administered into the living body together with the rod cells in order to induce the activation of the rod cells during immunotherapy, and is included as one component of the pharmaceutical composition. It may be. Here, the diseases targeted by immunotherapy are not particularly limited. For example, cancers such as colon cancer and stomach cancer, myeloma, liver cancer, leukemia, melanoma, prostate cancer, breast cancer, uterine cancer, Cancers or malignant tumors such as lung cancer, oral cancer, brain tumors; or chronic hepatitis due to hepatitis B or C virus, infections such as HIV, influenza, etc. The auxiliary active agent of the present invention may contain various commonly used ingredients, and the dosage thereof depends on the route of administration, the patient's body shape, age, physical condition, degree of disease, elapsed time after onset, etc. Can be selected as appropriate. For example, the auxiliary activator of the present invention can be carried out in the same embodiment as the above-described pharmaceutical composition of the present invention.

[0024] According to still another aspect of the present invention, there is provided a food composition comprising the above rod-shaped cell activator. The food composition of the present invention includes a liquid beverage such as a functional beverage. In addition to being used as a functional food, the food composition can be used as a quasi-drug, a component such as food or drink, or a food additive. In addition, the food composition in the present specification can be used as a functional food as it is, and can also be used as a food and drink, a pharmaceutical product, a quasi-drug, a component of food and drink, a food additive, and the like. The use enables daily and continuous ingestion of food, beverage, food composition or composition for oral intake having a rod-like cell activation effect and an immunostimulatory effect, and an effective rod-cell activation effect. It is possible to improve the constitution by immunostimulation effect, treat diseases such as cancer, malignant tumors and infectious diseases and prevent their onset. Examples of food compositions, foods or beverages containing the rod-like cell activator or immunostimulatory agent of the present invention include functional foods and health foods having rod-like cell activation effects or immunostimulatory effects. , General foods (juice, confectionery, processed foods, etc.), nutritional supplements (nutrient drinks, etc.). The food or beverage in this specification is not limited, but includes inorganic components such as iron and calcium, various vitamins, dietary fibers such as oligosaccharides and chitosan, proteins such as soybean extract, It can contain lipids such as tin, sugars such as sucrose and lactose, and plant extracts such as shiitake mushrooms.

 The invention's effect

 [0025] As shown in the following examples, the rod-shaped cell activator of the present invention has an action of promoting maturation of immature rod cells and an immunostimulatory action resulting from the action. Therefore, the present invention provides an effective means for treating and / or preventing cancer, malignant tumors and infectious diseases caused by immunostimulation.

 Furthermore, the present invention provides a method for inducing mature rod cells by treating immature rod cells. Since the shiitake mycelium extract also has an antioxidant effect, the method of the present invention can efficiently activate rod-shaped cells in an environment with little oxidative stress.

 Brief Description of Drawings

 [0027] [Fig. 1] An example of test results comparing the ability of shiitake mycelium extract and PSK to activate rod-shaped cells.

 FIG. 2 is an example of the results of microscopic observation of rod-shaped cells activated by shiitake mycelium extract and PSK supplementation.

 Example

 [0028] The following is a detailed explanation of preferred embodiments of the present invention. The present invention is not limited to these embodiments.

 [Example 1] Preparation of shiitake mycelium extract

After pure water was appropriately contained in a solid medium having 90 parts by weight of bagasse and 10 parts by weight of rice bran, shiitake mycelium was inoculated and allowed to stand in a culture chamber in which the temperature and humidity were adjusted to grow the mycelium. After the mycelium spread on the solid medium, the bagasse-based fibrin was bunched so that the amount passed through the 12 mesh was 24% by weight or less. Purified cellulase 2. Og was made into a mixture containing caustic medium while adding 3.5 L of pure water to 1. Okg of the unbundled medium and keeping at 40 ° C. Next, while the medium-containing mixture was circulated by a geared gear pump with speed change, the crushing and crushing action on the gear part was added to the solid medium for about 200 minutes, so that about 80% by weight of bagasse fiber was equivalent to the passage of 12 mesh. . Grinding and grinding media-containing mixture The smut was performed while gradually raising the temperature of the mixture. Thereafter, the medium-containing mixture was further heated to 90 ° C. for 30 minutes to inactivate the enzyme and sterilized. The obtained medium-containing mixture was filtered using a 60 mesh filter cloth and concentrated, and then a freeze-dried powder was obtained as a shiitake mycelium extract (LEM).

[Example 2] Measurement of human rod-shaped cell activation effect of shiitake mycelium extract

 (2-1) Induction of immature rod cells from human peripheral blood

Collect healthy human peripheral blood and use the RosetteSep Humon monocyte Enrichment cocktail kit (Stemcell) to prepare monocytes according to the protocol attached to the kit, and add 5xl0 ⁵ cells to RPMI1640 medium (sigma, containing 10% FBS). The cell suspension was prepared so as to be / ml. 500 L of the prepared cell suspension was dispensed into a 24-well plate, and further added to each of GM-CSF (Peprotech) and IL-4 (Peprotech) at 40 ng / ml. The cells were cultured at 37 ° C and 5% CO for 3 days. After 3 days, add 300 μL of culture supernatant to the cells.

2

Carefully blotted and added fresh RPMI 1640 medium (sigma, containing 10% FBS) 300 / z L. Further, GM-CSF (Peprotech) and IL-4 (Peprotech) were added at 40 ng / ml, and further cultured for 3 days (6 days in total) to induce immature rod cells. Induction of immature rod cells is a protein-specific antibody that expresses the expression of CD14 protein, a monocyte marker on the cell surface, and the expression of costimulatory factors, CD40, CD80, and CD86. Anti-CD14 antibody FITC labeling, anti-CD40 antibody FITC labeling, anti-CD80 antibody PE labeling, and anti-CD86 antibody PC5 labeling (Peprotech) were measured with a flow cytometer (Beckman Coulter Epics XL) for confirmation.

 (2-2) Induction of mature rod cells

 To immature rod cells prepared as described above, shiitake mycelium extract and PSK were added to a final concentration of 200 μg / ml, and cultured for 2 days at 37 ° C, 5% CO. . Also, the control

 2

The cells were cultured for 2 days at 37 ° C and 5% CO without adding anything. 2 days later, culture under each condition

 2

Cell surface costimulatory factors CD40, CD80, CD86 protein and mature rod cell specific marker CD83 protein expression levels of anti-CD40 antibody FITC label, anti-CD80 antibody PE label, anti-CD86 antibody PC5 label ( Measurement was performed with a flow cytometer (Beckman Coulter Epics XL) using Peprotech and anti-CD83 antibody FITC labeling (Peprotech). As a result, the expression level of CD40, CD80, CD83, and CD86, which were high in both the shiitake mycelium extract and PSK, was found to be higher than that in the control. The results are shown in Figure 1.

 [0030] As a result of the measurement, when either shiitake mycelium extract or PSK was added, an increase in the expression level was observed for all markers compared to the control mouth, and shiitake mycelium extract and It was confirmed that the addition of PSK activated the rod cells and promoted maturation. In addition, the expression level of the system in which shiitake mycelium was added for all the markers exceeded the expression level of the PSK system, and in particular, in the system in which the extract of shiitake mycelium of CD83, which is a matured rod-specific marker, was added. The expression level was confirmed to be more than twice that of the system to which PSK was added. From this, it was suggested that the rod-shaped cell activator of the present invention has a higher activity than the conventionally known rod-shaped cell activator.

 [0031] The mature rod-shaped cells obtained by the above procedure were observed with a microscope (x400 times). Figure 2 shows the results.

 It was confirmed that the mature rods induced by shiitake mycelium contained cells with greatly expanded cells compared to mature rods induced by PSK. From this, it was suggested that the rod-shaped cell activator of the present invention has a higher T cell interaction activity than the conventionally known rod-shaped cell activator.

Claims

The scope of the claims  [I] A rod-shaped cell activator comprising a shiitake mycelium extract.  [2] The rod-shaped cell activator according to claim 1, wherein maturation of the rod-shaped cells is promoted by activation of the rod-shaped cells.  [3] The rod-like cell activator according to claim 1 or 2, wherein the rod-like cell activity enhances the ability of the rod-like cells to produce interleukin 12, interferon γ or interleukin 1.  [4] A pharmaceutical composition comprising the rod-shaped cell activator according to any one of claims 1 to 3. [5] A pharmaceutical composition comprising the rod-like cell activity agent according to any one of claims 1 to 3, which is used for immunostimulation.  [6] A pharmaceutical composition comprising the rod-shaped cell activator according to any one of claims 1 to 3, for treating or preventing cancer or malignant tumor. [7] A pharmaceutical composition comprising the rod-shaped cell activator according to any one of claims 1 to 3, for treating or preventing an infectious disease. [8] A food composition comprising the rod-shaped cell activator according to any one of claims 1 to 3. [9] A method of inducing mature rod cells by treating immature rod cells using the rod cell activator according to any one of claims 1 to 3. [10] A mature rod-shaped cell obtained by treating an immature rod-shaped cell with the rod-shaped cell activator according to any one of claims 1 to 3.  [I I] A pharmaceutical composition comprising the mature rod-shaped cell according to claim 10.  [12] The pharmaceutical composition according to claim 11, for preventing or treating cancer, malignant tumor or infectious disease.  [13] An auxiliary activator for use in immunotherapy, comprising the rod-shaped cell activator according to claim 11.