Immune Cell Induction Device for Tumor Tissue

Abstract

To provide an immune cell induction apparatus to tumor tissue that is used in treatment / degradation prevention / the prevention of a tumor easily and effectively in a short time, and is simple and highly versatile, in which low temperature barometric pressure plasma is injected to noninvasively induce an immune cell to the tumor and activate local immunity, and the apoptosis in the tumor lesion is indirectly induced over a wide range in a low temperature barometric pressure plasma irradiation region and its vicinity.SOLUTION: An immune cell induction apparatus 1 to a tumor tissue has: a gas flow path 11 for conducting gas 51 in the tubular dielectric 13; a tubular intermediate electrode 12b around the outside of the tubular dielectric 13 as well as an upstream side grounding electrode 12a and a downstream side grounding electrode 12c; a power supply 31 applied to generate plasma at room temperature under conduction of the gas and under barometric pressure among those electrodes 12b and 12a / 12b; and a plasma airflow outlet 15 for injecting a plasma airflow 53 generated by the plasma to the tumor tissue 62 at the tip of the gas flow path 11. The immune cell is induced to the tumor tissue 62 with the plasma airflow 53.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to an immune cell induction device for tumor tissue, which can non-invasively induce immune cells against tumors and activate local immunity by injecting cryogenic atmospheric pressure plasma into tumor tissue, and can be used for the treatment, prevention of deterioration, and prevention of tumors.

Background Art

[0002] Tumors, especially malignant tumors that are prone to metastasis, such as gastric cancer, lung cancer, colorectal cancer, breast cancer, etc., are the leading cause of death among Japanese people. For the treatment of such tumors, there are surgical resection therapies for tumor primary foci and lymph nodes, such as sentinel lymph nodes where micrometastatic cells are metastasizing or about to metastasize from the breast cancer primary focus, not only the tumor primary focus but also other organs with metastasis, chemotherapy such as anticancer drugs and molecular target drugs, endocrine therapy such as hormone therapy, radiotherapy using radiation such as X-rays, electron beams, proton beams, heavy particle beams, α-rays, β-rays, γ-rays, or combined treatment using drug therapy and surgery.

[0003] Surgical resection therapy causes significant physical and mental damage to the human body. Chemotherapy is likely to cause severe side effects such as nausea, diarrhea, and hair loss due to damage not only to tumor cells but also to normal cells. Endocrine therapy is likely to cause side effects such as hot flashes and joint and muscle disorders. Radiation therapy has the risk of late-stage disorders such as taste disorders, respiratory disorders, rectal bleeding, and lymphedema in addition to acute disorders such as bone marrow suppression and rash.

[0004] In recent years, cryogenic atmospheric pressure plasma technology has been tried as a new tumor treatment method. For example, in Patent Document 1, there are a pair of opposing electrodes with recesses formed on the opposing surfaces of the electrodes, a housing covering the plasma generation region between the pair of opposing electrodes, a gas inlet for introducing a gas for generating plasma into the housing, and an irradiation unit for irradiating the plasma generated by the pair of opposing electrodes to the outside of the housing. The pair of opposing electrodes has a plasma density of 1×10 14 cm -3 or more and 1×10 17 cm -3It generates plasma within the following range, and the irradiation unit irradiates the ovarian tumor cells with the plasma. An ovarian cancer treatment device is disclosed. Such an ovarian cancer treatment device is such that ovarian cancer cells die exclusively at the low-temperature atmospheric pressure plasma irradiation site due to direct apoptosis by the low-temperature atmospheric pressure plasma.

[0005] Also, instead of directly irradiating tumor cells with plasma, an antitumor effect has been found in the irradiated solution. For example, in Patent Document 2, in an antitumor aqueous solution that selectively kills tumor cells, a first aqueous solution in which a solute containing at least one of disodium hydrogen phosphate, sodium hydrogen carbonate, L-glutamine, L-histidine, and disodium L-tyrosine dihydrate is dissolved in water is irradiated with atmospheric pressure plasma to obtain a second aqueous solution, and an antitumor aqueous solution that is the frozen product of the second aqueous solution is disclosed.

[0006] Furthermore, Patent Document 3 discloses an R 1 -NHCH2COCH2CH2COOR 2 A pharmaceutical composition for enhancing the therapeutic effect of cancer or tumor by plasma therapy, which contains a compound represented by the formula, or a salt or ester thereof, is disclosed. Patent Document 3 discloses a direct cytotoxic effect on tumors by direct plasma irradiation using this pharmaceutical composition.

[0007] However, it is not known to induce local immunity at the tumor, especially the primary tumor focus and metastatic sites therefrom, in the local area or slightly deep area irradiated with plasma.

[0008] Also, Non-Patent Document 1 analyzes the immunity of the surface skin by low-temperature atmospheric pressure plasma irradiation. This Non-Patent Document 1 mainly examines skin damage by low-temperature atmospheric pressure plasma irradiation, observes inflammatory cell infiltration in the skin at the irradiation site, and causes a direct inflammatory reaction in the skin at the irradiation site as an adverse event. However, it is rather the induction of an inflammatory reaction due to a burn (96°C) at the irradiation site than the induction of tumor immunity or immune induction having a therapeutic effect.

Prior Art Documents

Patent Documents

[0009]

Patent Document 1

Patent Document 2

Patent Document 3

Non-Patent Documents

[0010]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0011] The present invention has been made to solve the above problems, and by injecting low-temperature atmospheric pressure plasma into a tumor, non-invasively inducing immune cells to activate local immunity, and indirectly inducing apoptosis in tumor lesions not only at the site irradiated with low-temperature atmospheric pressure plasma but also in a wide range in the vicinity thereof, it is an object of the present invention to provide a simple and highly versatile immune cell induction device for tumor tissue that can be effectively used for the treatment, prevention of deterioration, and prevention of tumors simply and in a short time.

Means for Solving the Problems

[0012] An immune cell induction device for tumor tissue made to achieve the above object includes a gas flow path for conducting gas in a glass tube or a quartz tube which is a tubular dielectric, a tubular middle electrode surrounding the outside of the tubular dielectric, and an upstream ground electrode disposed upstream of the middle electrode and At a distance of 70 mm a downstream ground electrode disposed downstream of the middle electrode and At a distance of 40 mmA configured downstream ground electrode, a power source applied to generate plasma at room temperature under atmospheric pressure with the gas conducting between these electrodes, and one or more plasma airflow nozzles for directly and / or indirectly injecting the plasma airflow generated by the plasma to the tumor tissue at the tip of the gas flow path. By doing so It is for inducing immune cells in the tumor tissue with the plasma.

[0013] In this immune cell induction device for tumor tissue, it is preferable that the plasma airflow nozzle has a diameter of 1 mm or less, which can enhance the production efficiency of reactive oxygen and nitrogen species, induce the immune cells, and activate local immunity. In previous experiments, high effects were obtained when the diameter of the plasma airflow nozzle was 200 - 700 μm.

[0014] In this immune cell induction device for tumor tissue, the gas flow rate should be 1.5 L / min or less, preferably 0.5 - 1.5 L / min, so as not to dry the living tissue. For example, when the diameter of the plasma airflow nozzle is 650 μm, the flow rate is 1 L / min.

[0015] In this immune cell induction device for tumor tissue, the plasma is, for example, limited such that the average current flowing from the plasma to the tumor tissue is 400 μA or less.

[0016] Examples of the gas used in this immune cell induction device for tumor tissue include helium gas, argon gas, nitrogen gas, oxygen gas, or a mixed gas of any of these.

[0017] In this immune cell induction device for tumor tissue, it is preferable that the tip is tapered and / or the plasma airflow nozzle has a diameter smaller than the diameter of the tubular dielectric, and it is further preferable to limit the average current flowing from the plasma airflow to the tumor tissue to 400 μA or less.

[0018] The immune cell induction device for this tumor tissue is a dielectric barrier discharge in which plasma is generated in a glass or quartz tube. The generation of plasma by the dielectric occurs intermittently like pulsed discharge plasma. By providing a complete dielectric barrier (a structure without electrodes in the discharge space) where all electrodes are installed outside the dielectric, it is more preferable that the density of the plasma, although relatively low, can minimize the influence of heat.

[0019] The immune cell induction device for this tumor tissue may be one in which the plasma airflow outlet is provided at the tip of an endoscope, the tip of a catheter, or the tip of a probe.

[0020] This immune cell induction device for tumor tissue injects a plasma airflow into the tumor tissue in one shot or multiple shots, either unidirectionally or multidirectionally. This immune cell induction device for tumor tissue uses a switching device to output a high-voltage power supply for injecting the one shot or the multiple shots. This immune cell induction device for tumor tissue is such that the tumor tissue is, in vivo, a primary tumor focus, tumor cells, micro-tumor cells, superficial lymph nodes, sentinel lymph nodes, lymph nodes, blood vessels, disseminated tumor tissue, and / or metastatic tissue. This immune cell induction device for tumor tissue is for locally injecting the plasma airflow to increase B lymphocytes, macrophages, neutrophils, T cells, NK cells, dendritic cells, and / or plasma cells in the tumor tissue. The method of using the immune cell induction device for this tumor tissue, which is made to achieve the above object, is to inject the plasma airflow in one shot or multiple shots in the to one direction or in multiple directions towards the tumor tissue.

[0021] The method of using the immune cell induction device for this tumor tissue is, for example, to use the output of a high-voltage power supply through a switching device to perform the one-shot or multiple shots, injecting for several seconds to ten-odd minutes per shot.

[0022] The method of using the immune cell induction device for this tumor tissue is preferably such that the tumor tissue is, in vivo, a primary tumor focus, tumor cells, micro-tumor cells, superficial lymph nodes, sentinel lymph nodes, lymph nodes, blood vessels, disseminated tumor tissue, and / or metastatic tissue.

[0023] The method of using the immune cell induction device for this tumor tissue is to locally inject the plasma airflow to increase B lymphocytes, macrophages, neutrophils, T cells, NK cells, dendritic cells, and / or plasma cells in the tumor tissue.

Advantages of the Invention

[0024] The immune cell induction device for tumor tissue of the present invention can non-invasively induce immune cells in the tumor tissue by injecting a plasma airflow generated by low-temperature atmospheric pressure plasma at room temperature (direct plasma irradiation), thereby activating local immunity.

[0025] By using this immune cell induction device for tumor tissue, through the injection of a plasma airflow generated by low-temperature atmospheric pressure plasma in the tumor tissue of the living body, that is, in vivo, instead of the direct bactericidal effect or direct apoptosis induction of tumor cells by conventional plasma direct irradiation, an indirect apoptosis induction effect can be achieved through an immune reaction mediated by immune cell induction, thereby expressing an action mechanism completely different from that of conventional plasma devices.

[0026] By using this immune cell induction device for tumor tissue, it can be used for the treatment, prevention of deterioration, and prevention of tumors of tumor tissue, especially the primary tumor nest and micro-tumor cells that are metastasizing, as well as metastatic or metastasizing superficial lymph nodes, sentinel lymph nodes, lymph nodes, blood vessels, disseminated tumor tissues, and metastatic tissues such as metastatic organs, in a simple and short time, safely and effectively, without damaging normal tissues.

[0027] This immune cell induction device for tumor tissue can increase immune cells, especially B lymphocytes, macrophages, neutrophils, T cells, NK cells, dendritic cells, and / or plasma cells in the tumor tissue by locally injecting the plasma airflow into the tumor tissue. As a result, it can induce necrosis of tumor cells in the tumor tissue, thereby achieving the treatment, prevention of deterioration, and prevention of tumors.

[0028] According to this immune cell induction device for tumor tissue, even without injecting a plasma airflow throughout the entire tumor tissue, by irradiating a part of the tumor tissue with a plasma airflow for a predetermined time, immune cells can be induced to activate local immunity, and as a result, the immune function of the patient himself / herself can be enhanced, thereby inducing necrosis of the tumor cells in the entire tumor tissue.

[0029] According to this immune cell induction device for tumor tissue, not only the tumor cells of the epidermal tumor tissue but also the tumor cells of the subcutaneous tumor tissue can be induced to necrosis, so non-invasive treatment can be performed.

[0030] This immune cell induction device for tumor tissue is not only simple and highly versatile, but also excellent in portability, being small and portable, and can be easily used at various medical sites without being a complex and large-scale device.

[0031] Since this immune cell induction device for tumor tissue has a simple structure, it can be manufactured inexpensively and easily.

[0032] According to the method of using this immune cell induction device for tumor tissue, immune cells can be induced and the tumor tissue can be indirectly induced to necrosis, and it can be used for tumor treatment with almost no side effects.

Brief Description of the Drawings

[0033]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Embodiments for Carrying Out the Invention

[0034] Hereinafter, embodiments for carrying out the present invention will be described in detail, but the scope of the present invention is not limited to these embodiments.

[0035] As shown in FIG. 1 which is a schematic diagram thereof, the immune cell induction device 1 for a tumor tissue of the present invention includes a low-temperature atmospheric pressure plasma generation device 10, a gas supply device 20 having a gas cylinder 23, a voltage application device 30 for generating plasma 52, and a plasma generation control device 40 which is a personal computer (PC) for controlling the supply of carrier gas 51 and the generation of plasma 52.

[0036] The immune cell induction device 1 for this tumor tissue can suppress the drying of the tumor tissue by narrowing the plasma air flow outlet 15 to reduce the gas flow rate flowing into the tumor tissue to 0.5 to 1.5 L / min. Furthermore, by narrowing the outlet, the current flowing into the tumor tissue can also be suppressed. As a result, thermal damage due to Joule heat of the tumor tissue and the surrounding normal tissue can be suppressed.

[0037] The low-temperature atmospheric pressure plasma generation device 10 is for ejecting a plasma air flow 53 generated by the generation of plasma 52. In the low-temperature atmospheric pressure plasma generation device 10, a tubular dielectric 13 made of glass or quartz is provided as an inner tube dielectric forming a gas flow path 11. At the base side of the tubular dielectric 13, it is connected to a gas cylinder 23 via a flexible gas pipe 21, an on-off valve 22a, a pressure regulator 22b, and a gas flow rate regulator 22c. The other tip side of the tubular dielectric 13 is covered while surrounding the tubular dielectric 13 with a nozzle 16 made of a dielectric such as polytetrafluoroethylene and having only a plasma air flow outlet 15 open. A plasma air flow outlet 15 for ejecting a plasma air flow 53 having a diameter smaller than the diameter of the tubular dielectric 13 is opened in the nozzle 16.

[0038] An outer cylinder case 14, which is an outer cylinder dielectric made of cylindrical polytetrafluoroethylene (Teflon; registered trademark) provided with a gap so as to surround the gas flow path 11, is provided on the tubular dielectric 13. A cylindrical coaxial middle electrode 12b is wound around so as to be in contact with the tubular dielectric 13. An upstream side electrode 12a, which is cylindrical and coaxial, is wound around the upstream side of the tubular dielectric 13 so as to be in contact with the outer cylinder case 14. Also, a downstream side electrode 12c, which is cylindrical and coaxial, is wound around the downstream side of the tubular dielectric 13 so as to be in contact with the nozzle 16. The distance between 12a and 12b is 70 mm, and the distance between 12b and 12c is 40 mm. The thickness of the glass tube is 1 mm.

[0039] The electrode 12, which consists of the middle electrode 12b, the upstream grounding electrode 12a, and the downstream grounding electrode 12c, is connected to the high-voltage power supply 31. The upstream grounding electrode 12a and the downstream grounding electrode 12c are grounded.

[0040] When an alternating current or pulsed voltage is applied between the electrodes 12 by the high-voltage power supply 31, dielectric barrier discharge occurs, resulting in the generation of plasma 52. Due to the flow of the carrier gas 51 in the gas flow path 11 by the plasma 52, the plasma airflow 53 is ejected from the plasma airflow ejection port 15 toward the tumor region 60 of the patient.

[0041] This immune cell induction device 1 for the tumor tissue has a bifurcated structure that separates the generation of stable plasma between the upstream grounding electrode 12a and the middle electrode 12b and the downstream side (between the middle electrode 12b and the downstream grounding electrode 12c) for supplying the plasma airflow. Therefore, it can generate and maintain stable plasma even under a lower gas flow rate than conventional devices that generate two electrodes and single-electrode plasma. Furthermore, by providing the plasma airflow ejection port 15, the supply amount of charged particles and excited species contained in the plasma airflow can be controlled.

[0042] The tubular dielectric 13 is a tubular one made of dielectric, which may be, for example, a glass tube or a quartz tube. The plasma airflow ejection port 15 is provided, for example, at the tip side of the tubular dielectric 13 and is blocked by the nozzle 16, and a small-diameter ejection hole is provided coaxially with the tubular dielectric 13 in the center of the nozzle 16. The plasma airflow ejection port 15 has a diameter of 650 μm. An appropriate diameter that does not dry the biological tissue with plasma irradiation for up to about 15 minutes is about several hundred μm. If the diameter is too large beyond this range, a large amount of carrier gas is required to maintain the plasma stably, and the biological tissue is likely to dry. Furthermore, as the plasma airflow ejection port 15 becomes larger, the current flowing into the biological tissue also increases. As a result, the biological tissue is damaged by Joule heat.

[0043] The plasma gas outlet 15 is shown in Fig. 1 as an example of being opened in the nozzle 16 that closes the tip of the tubular dielectric 13. However, as shown in Fig. 2(a), the tip of the tubular dielectric 13 with the same diameter may also serve as the plasma gas outlet 15 without using a nozzle. The plasma gas outlet 15 may be a single hole where the tip of the tubular dielectric 13 is closed in a dome shape or tapered as shown in Fig. 2(b). The plasma gas outlet 15 may be a plurality of holes opened in the nozzle 16 as shown in Fig. 2(c). The plasma gas outlet 15 may be one opened at the tapered tip of the conical or dome-shaped nozzle 16 that closes the tip of the tubular dielectric 13 as shown in Fig. 2(d).

[0044] As shown in Fig. 2(e), the nozzle 16 may be provided with a supporter 16a so as to maintain a predetermined distance from the plasma gas outlet 15 to the surface of the tumor region 60. Specifically, as shown in Fig. 3, four claw-shaped supporters 16a extend from a nozzle made of polytetrafluoroethylene, and the tips of the four supporters 16a may be bent inward. The supporter 16a is preferably adjusted so that the distance from the plasma gas outlet 15 to the surface of the tumor region 60 is 10 to 20 mm according to the gas flow rate and the length of the plasma jet.

[0045] The nozzle 16 as shown in Figs. 2(c) to 2(e) can be appropriately replaced as needed. For example, as shown in Fig. 3, it may be replaced with a nozzle 16 in which the diameter and shape of the nozzle 16, the diameter of the plasma gas outlet 15, and the length of the supporter 16a are appropriately adjusted.

[0046] Although not shown in the figure, it may be a single hole where only the inner diameter of the tubular dielectric 13 tapers while the outer diameter remains constant and the tip is open, or a single hole where the entire tubular dielectric 13 tapers and the tip is open, or a single hole where the tip is open because the tubular dielectric 13 itself is a thin tube with the same diameter, or a plurality of holes with a number of holes opened at the tip of the tubular dielectric 13, or an elliptical shape or groove shape within the range of the said diameter.

[0047] The voltage application device 30 has an AC power source or a pulse power source as the high-voltage power source 31, and applies a pulse voltage of 1 to 10 kV, preferably about several kV to 10 kV, at a low frequency of several to several tens of kHz, preferably about 10 kHz, to the electrode 12. When applying to the electrode 12 while flowing a carrier gas through the tubular dielectric 13, a plasma airflow 53 is generated and ejected as a non-equilibrium plasma jet in which the electron and molecular temperatures are different.

[0048] By using the dielectric barrier discharge method, the generation of plasma can be made intermittent. The current (discharge current) of the plasma airflow blown out from the plasma airflow outlet 15 is shown in FIG. 4. The intermittent plasma with a pause period reacts with oxygen, nitrogen, and moisture in the air to generate active oxygen and nitrogen species, and can induce immune cells into the tumor tissue. Furthermore, by using the intermittent plasma with a pause period, an increase in the temperature of the plasma airflow can be suppressed. Even when using the dielectric barrier discharge method, at a frequency of 100 kHz or more, the pause period of the plasma is not sufficient, and it is difficult to suppress overheating of the plasma airflow.

[0049] The voltage application device 30 can generate the plasma by the dielectric barrier discharge method at a high voltage (10 kV) and a low current (1 mA or less). The average current value obtained from the current values shown in FIG. 4 is about 400 μA for both the positive current and the negative current. The maximum current value is 1 mA or less. As a result, the average power input is about 1 W. Since the power input for plasma generation is small and the current flowing to the surface of the tumor region 60 is small, overheating of the plasma airflow can be suppressed, Joule heating of the tumor region can be suppressed, and thermal damage can be avoided.

[0050] The gas supply device 20 supplies the carrier gas 51 from the gas cylinder 23. The carrier gas 51 includes helium gas, argon gas, nitrogen gas, oxygen gas, and / or a mixed gas thereof. However, in terms of the ease of chemical reactions due to entraining the surrounding atmosphere (oxygen, nitrogen, moisture) and the requirement of less input power for plasma generation, helium is more preferable. The highly reactive active species (oxygen and nitrogen species) generated from atmospheric pressure plasma are transported to the surface of the tumor region 60 by the plasma airflow 53.

[0051] The gas supply device 20 is controlled by the plasma generation control device 40 to adjust the flow rate of the carrier gas from the gas cylinder 23 to the tubular dielectric 13 by the on-off valve 22a, the pressure regulator 22b, and the gas flow rate regulator 22c. The flow rate of the carrier gas is preferably set to 0.5 - 1.5 L / min. If it is less than this range, the flow rate of the plasma airflow 53 is too small to sufficiently transport the generated active oxygen and nitrogen species to the surface of the tumor region. If it is more than this range, the surface of the tumor region will be excessively dried by the plasma airflow 53, making it difficult to induce immune cells.

[0052] The plasma generation control device 40 has a central control unit (CPU) and an input terminal to control the supply (opening / closing and flow rate) of the carrier gas 51 from the gas cylinder 23 to the tubular dielectric 13 and the application of the high-voltage power supply 31 of the AC power supply or the pulse power supply of the voltage application device 30 for the generation of the plasma 52. The CPU is for controlling according to the signal from the input terminal and has a carrier gas control circuit for controlling the supply of the carrier gas 51 and a voltage application control circuit for controlling the voltage, current, and pulse of the high-voltage power supply 31 of the AC power supply or the pulse power supply of the voltage application device 20 (not shown).

[0053] The immunocyte induction device 1 for tumor tissue, as shown in FIG. 1, may directly hold the tubular dielectric 13 and eject the plasma airflow 53 directly to the tumor tissue 62 or indirectly to the tumor tissue 62 through the epidermis 61. Instead of directly holding the thin tubular dielectric 13, it may be provided at the tip of the endoscope 17a as shown in FIG. 5(a), at the tip of the catheter 17b as shown in FIG. 5(b), or at the tip of the probe 18 having a plurality of circular or grooved plasma airflow ejection ports 15 at the tip as shown in FIG. 5(c). As shown in FIG. 5(a-2), the endoscope or catheter may be provided with a resin housing case 19 for easy gripping.

[0054] Also, in the method for generating atmospheric pressure plasma in the immunocyte induction device 1 for tumor tissue, a stable dielectric barrier discharge is generated inside the tubular dielectric 13 by the electrode 12 provided outside the tubular dielectric 13, and the glow-like discharge plasma airflow 53 can be blown out from the plasma airflow ejection port 15.

[0055] This immunocyte induction device 1 for tumor tissue is used as follows.

[0056] When the immunocyte induction device 1 for tumor tissue is driven and a predetermined voltage is applied between the electrodes 12 with a predetermined sine wave or pulse wave while flowing the carrier gas 51, plasma 52 is generated by dielectric barrier discharge, and the plasma airflow 53 generated by the generation of the plasma 52 is ejected from the plasma airflow ejection port 15. This plasma airflow 53 is sprayed and irradiated directly to the tumor tissue 62 or indirectly to the tumor tissue 62 through the epidermis 61 to the tumor region 60.

[0057] As a method for treating tumors using this immunocyte induction device 1 for tumor tissue, the plasma airflow 53 is irradiated to the tumor region 60 for 1 second to 1 hour, preferably 1 to 20 minutes, more preferably 10 to 15 minutes. The plasma airflow 53 may be evenly sprayed and irradiated to the tumor region 60 of a tumor-afflicted patient or tumor-afflicted animal in vivo, or may be concentrated at one location.

[0058] The method for treating tumors using the immune cell induction device 1 for this tumor tissue involves performing single-shot or multi-shot treatments once to several times a day, preferably 1 to 3 times, either only once, daily, or at intervals of 1 to 7 days or 1 to 4 weeks, continuously for 1 day to about 1 month until the tumor tissue disappears or decreases, or until the condition of the tumor is alleviated.

[0059] Such tumor tissue is preferably the primary tumor focus, but it may also be tumor cells of the primary tumor focus, or it may be the primary tumor focus and lymph nodes to be dissected. It may be the sentinel lymph node that first receives lymph flow from the primary tumor focus and where lymphogenous micrometastasis can first occur, or it may be lymph nodes within the sentinel lymph node that act on micrometastatic tumor cells (micrometastatic tumor cells) and the lymphatic vessels and axillary lymph nodes closest to it, or it may be superficial lymph nodes, or blood vessels, or tissues to which the tumor has metastasized.

[0060] According to the method for treating tumors using the immune cell induction device 1 for this tumor tissue, immune cells can be induced within the tumor tissue. The immune cells are, for example, B lymphocytes (B cells), macrophages, neutrophils, T cells, NK cells, dendritic cells, and / or plasma cells. Specifically, B lymphocytes (B cells) such as B lymphocytes that are precursor cells of antibody-producing cells and cooperate with helper T cells to produce antibodies, and memory B cells that are stimulated by helper T cells and remember antigen information; Macrophages that are rich in cytoplasm, easily adhere, extend pseudopods and move, and transmit antigen (tumor) information to helper T cells; Neutrophils, which are a type of white blood cell, have strong phagocytic activity, extend pseudopods to take in tumor cells, and kill tumor cells by the action of enzymes; T cells such as helper T cells that produce cytokines to activate B lymphocytes and assist in antibody production together with B lymphocytes, killer T cells that kill and remove tumor cells under the control of helper T cells, and regulatory T cells that suppress the immune system from acting against itself and prevent immune abnormalities; NK cells (natural killer cells) that damage not only tumor cells but also cells recognized as abnormal; Dendritic cells that take up antigens such as tumor cells and transmit antigen information to T cells to initiate an immune response; Plasma cells, which are the final differentiated form of bone marrow-derived cells and secrete antibodies; and the like.

[0061] The plasma airflow 53 is ejected from the plasma airflow ejection port 15 and irradiated onto the tumor region 60, which is the epidermal 61 or tumor tissue 62. Since the temperature of the epidermal 61 or tumor tissue 62 during plasma irradiation is at most 39 - 47°C, the tumor region 60 does not cause inflammation and activate immune cells.

[0062] By inducing an increase and activation of immune cells by irradiating the plasma airflow 53 onto the tumor region 60, even if only a part of the tumor tissue 62 is irradiated without irradiating the entire tumor tissue 62, the detailed mechanism by which the tumor tissue 62 is induced to die is not necessarily clear, but is speculated as follows.

[0063] By irradiating the plasma airflow 53, infiltration of neutrophils is observed in a wide range of the tumor tissue 62, including the periphery of the tissue degeneration region. A large amount of infiltration of lymphocytes and macrophages is observed within the tumor nest. From this, it can be seen that by irradiating the plasma airflow 53, neutrophils can be migrated widely within the tumor tissue 62 of the subcutaneous tissue, and the infiltration of lymphocytes and macrophages into the tumor nest can be increased. As a result, by increasing immune cells, necrosis of tumor cells in the tumor tissue can be induced, and tumor treatment, prevention of deterioration, and prevention can be carried out by apoptosis.

Example

[0064] Hereinafter, examples using an immune cell induction device for tumor tissue to which the present invention is applied and comparative examples to which the present invention is not applied will be described in detail.

[0065] (Example 1) Male nude mice at 5 weeks of age (purchased from Japan SLC, Inc.) were heterotopically transplanted subcutaneously on the back with 253J-BV, a human bladder cancer cell line; 4×10 6 cells / 100 μl. For the nude mice bearing tumors created at 7 - 8 weeks of age, a plasma airflow generated by an immune cell induction device for tumor tissue to which the present invention is applied, with a 10 kV sine wave generated under the condition of a frequency of 33 kHz for helium gas at 1 L / min, was irradiated for 15 minutes. Thereafter, the fluorescence intensity was measured using an IVIS spectrum by a luminol reaction that emits light with reactive oxygen species (ROS) to examine the antitumor effect, and the temperature distribution of the epidermis was also examined using a FLIR T530 infrared thermographic camera. The results of the luminol reaction that emits light with reactive oxygen species (ROS) are shown by the graph in Fig. 6(a) and the photograph in Fig. 6(b-2). In Fig. 7, the upper graph shows the time change of the highest temperature in the distribution, and the lower photograph shows the results of the temperature distribution of the epidermis.

[0066] (Comparative Example 1) Instead of the plasma airflow generated by the immune cell induction device for tumor tissue, helium was irradiated in the same manner as in Example 1, except that a non-plasma-treated helium airflow was used without applying a voltage. Thereafter, it was similarly stained with luminol. The results are shown by the graph in Fig. 6(a) and the photograph in Fig. 6(b-1).

[0067] (Staining comparison results between Example 1 and Comparative Example 1) As is clear from Fig. 6, the reactive oxygen species were significantly increased by nearly about 4 times by the irradiation of the plasma airflow generated by the immune cell induction device for tumor tissue as in Example 1, compared with the case of irradiating with a helium airflow as in Comparative Example 1. From this, it was found that the immune cells were activated by the reactive oxygen generated at the irradiation site by the direct irradiation of low-temperature atmospheric pressure plasma using this immune cell induction device for tumor tissue and were induced from around the tumor mass into the tumor mass.

[0068] (Example 2) Subcutaneously on the back of male nude mice at 5 weeks of age (purchased from Japan SLC, Inc.) with 253J-BV, a human bladder cancer cell line; 4×10 6100 μl / individual was transplanted to create a cancer-bearing model mouse. In the tumor region of the created nude mouse, the plasma airflow generated by the immune cell induction device for tumor tissue to which the present invention is applied was directly irradiated from a distance of about 1.5 cm away for 15 minutes at a 10 kV sine wave under the condition of a frequency of 33 kHz with respect to 1 L / min of helium gas through the epidermis to the tumor tissue. It was stained with luminol in the same manner as in Example 1. Also, sections were cut out in the tumor region, and HE staining (hematoxylin and eosin staining) and TUNEL staining (TdT-mediated dUTP nick end labeling staining) for staining apoptotic tissues and cells were performed. The results of luminol staining are shown in (a), the results of HE staining are shown in (b-1) of the same figure, and the results of TUNEL staining are shown in (b-2) of the same figure.

[0069] (Staining results of Example 2) As is clear from FIG. 8, as shown in (a) of the same figure, it was shown that reactive oxygen species were present according to the luminol fluorescence measurement results. Also, as shown in (b-1) of the same figure, degenerated epithelium was formed between normal epithelia, but a part of the bladder cancer cells was necrotic and apoptotic like the dark part in (b-2) of the same figure by the irradiation of the plasma airflow. As shown by the black frame in (c) of the same figure, immune cells and exudate increased in the surface layer of the plasma irradiation site. From this, B lymphocytes, macrophages, and neutrophils accumulated in the tumor tissue and the number of immune cells increased. From this, it was found that the immune cells were activated by apoptosis generated by plasma irradiation and accumulated at that site.

[0070] (Example 3) In the same manner as in Example 2, in the tumor region of the nude mouse, the plasma airflow generated by the immune cell induction device for tumor tissue to which the present invention is applied was directly irradiated from a distance of about 1.5 cm away for 15 minutes at a 10 kV sine wave under the condition of a frequency of 33 kHz with respect to 1 L / min of helium gas through the epidermis to the tumor tissue. For the subcutaneous tissue and the tumor tissue, as immune cells, B lymphocytes and neutrophils were HE-stained and counted under a microscope, and macrophages were counted by staining with CD68 (KP-1).

[0071] (Comparative Example 2) B lymphocytes, macrophages, and neutrophils were counted in the same manner as in Example 3, except that the epidermis of the tumor site was cauterized with an electrosurgical knife instead of irradiating the tumor tissue with a plasma airflow using an immune cell induction device for tumor tissue.

[0072] (Comparison Results of Pathological Analysis of Immune Cells between Example 3 and Comparative Example 2) For the plasma airflow irradiation treatment of Example 3 and the electrosurgical knife treatment of Comparative Example 2, the number of B lymphocytes, macrophages, and neutrophils per unit area in the subcutaneous tissue and tumor tissue was compared. The results are shown in Fig. 9.

[0073] As is clear from Fig. 9(a), in the subcutaneous tissue, the increase in macrophages was the main effect in the electrosurgical knife treatment, and only a slight increase in B lymphocytes and neutrophils was observed, while the increase in neutrophils was the main effect in the plasma airflow irradiation treatment, and only a slight increase in B lymphocytes was observed. On the other hand, in the tumor cells, the infiltration of B lymphocytes and macrophages was less in the electrosurgical knife treatment, while the infiltration of B lymphocytes and macrophages was more in the plasma airflow irradiation treatment.

[0074] From these results, it was suggested that, unlike the treatment group using an electrosurgical knife, in the plasma treatment group, lymphocytes and macrophages were induced in the tumor nests slightly deeper than the irradiation site, thereby necrotizing the tumor tissue.

Industrial Applicability

[0075] According to the immune cell induction device for tumor tissue to which the present invention is applied, and the method of using the same, By injecting atmospheric pressure plasma into the tumor tissue to induce immune cells and activating local immunity, it can be used for the treatment, prevention of deterioration, and prevention of tumors by necrotizing the tumor tissue through apoptosis.

Explanation of Signs

[0076] 1 is an immune cell induction device for tumor tissue, 10 is a low-temperature atmospheric pressure plasma generation device, 11 is a gas flow path, 12 is an electrode, 12a is an upstream ground electrode, 12b is a middle electrode, 12c is a downstream ground electrode, 13 is a tubular dielectric, 14 is an outer cylinder case, 15 is a plasma airflow jet outlet, 16 is a nozzle, 16a is a supporter, 17a is an endoscope, 17b is a catheter, 18 is a probe, 19 is a housing case, 20 is a gas supply device, 21 is a gas pipe, 22a is an on-off valve, 22b is a pressure regulator, 22c is a gas flow regulator, 23 is a gas cylinder, 30 is a voltage application device, 31 is a power supply, 40 is a plasma generation control device, 51 is a carrier gas, 52 is a plasma, 53 is a plasma airflow, 60 is a tumor region, 61 is an epidermis, and 62 is a tumor tissue.

Claims

1. A gas flow path for conducting gas within a glass tube or quartz tube which is a tubular dielectric, a tubular intermediate electrode surrounding the outside of the tubular dielectric, an upstream ground electrode disposed at a distance of 70 mm upstream of the intermediate electrode, and a downstream ground electrode disposed at a distance of 40 mm downstream of the intermediate electrode, a power source applied between these electrodes for generating plasma at room temperature under atmospheric pressure with the gas conducting, and one or more plasma airflow nozzles for directly and / or indirectly injecting the plasma airflow generated by the plasma to the tumor tissue at the tip of the gas flow path, and it is for inducing immune cells in the tumor tissue with the plasma, characterized in that it is an immune cell induction device for tumor tissue.

2. The immune cell induction device for tumor tissue according to claim 1, characterized in that the plasma airflow nozzle has a diameter of 200 to 700 μm, thereby inducing the immune cells and activating local immunity.

3. The immune cell induction device for tumor tissue according to any one of claims 1 to 2, characterized in that the gas flow rate is 0.5 to 1.5 L / min, thereby suppressing drying of the tumor tissue.

4. The immune cell induction device for tumor tissue according to any one of claims 1 to 3, characterized in that the gas is helium gas, argon gas, nitrogen gas, oxygen gas, or a mixed gas of any of these.

5. The immune cell induction device for tumor tissue according to any one of claims 1 to 4, characterized in that the tip is tapered and / or the plasma airflow nozzle has a diameter smaller than the diameter of the tubular dielectric.

6. The immune cell induction device for tumor tissue according to any one of claims 1 to 5, characterized in that the plasma airflow nozzle is provided at the tip of an endoscope, the tip of a catheter, or the tip of a probe.

7. The immune cell induction device for tumor tissue according to any one of claims 1 to 6, wherein the plasma airflow is injected into the tumor tissue in one shot or multiple shots in one direction or multiple directions.

8. The immune cell induction device for tumor tissue according to claim 1, wherein the tumor tissue is a primary tumor focus, tumor cells, micro-tumor cells, superficial lymph nodes, sentinel lymph nodes, lymph nodes, blood vessels, disseminated tumor tissue, and / or metastatic tissue in vivo.

9. The immune cell induction device for tumor tissue according to claim 1, which is used by locally injecting the plasma airflow to increase B lymphocytes, macrophages, neutrophils, T cells, NK cells, dendritic cells, and / or plasma cells in the tumor tissue.

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