JPH09103483A - Method for activating cell-mediated immunity by heat treatment of blood and apparatus therefor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for activating cell-mediated immunity(CMI) by heat treating blood without imparting physical denaturation to the blood cells and an apparatus therefor. SOLUTION: This method comprises activating the CMI of lymphocytes, etc. by heating the blood of a living body to 38 to 41 deg.C by a heat source of 55 to 80 deg.C. This device for activating the CMI by the heat treatment of the blood is provided with a blood pipeline 1 which takes the blood out of the living body and a heating mechanism 3 for heating the blood in a range of 55 to 80 deg.C and a blood delivery pump 2 for regulating the flow rate of the blood so that the blood is heated in a range of 38 to 41 deg.C heating temp. in correspondence to the heating temp. of this heating mechanism 3 in the middle of this blood pipeline 1.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and apparatus for activating cell-mediated immunity by heat treatment of blood.

[0002]

2. Description of the Prior Art Conventional major antitumor therapies include chemotherapy, radiation therapy and surgery. Since cancer cells and AIDS virus are generally considered to be vulnerable to heat, the temperature at the tumor site can be controlled by using a complicated and expensive device that generates a hot bath, ultrasonic waves, electromagnetic energy, laser, etc. There is hyperthermia that artificially raises the growth rate of tumors. Any usage is 40
It is a hyperthermia that heats the whole body and the local area in the temperature range of 42 to 42 ° C.

[0003]

However, the temperature of the hot water in the bathtub that we usually use is approximately 40 to 42 ° C., which is consistent with the temperature used in the above-mentioned hyperthermia, but skin cancer, etc. There is a problem that the conventional hyperthermia has not yet occupy a definite position as an effective treatment method due to the fact that there is no clinical case that the onset of skin cancer has been cured and the skin cancer that has developed has been cured. .
As a result of actually examining various heat conditions of blood, it was confirmed that AIDS virus and cancer cells were not killed under the above-mentioned heat conditions. In addition, even if a certain effect is recognized, the equipment to be used is expensive, so the number of facilities to install is limited to a very small number, so that many users cause inconveniences such as transportation and enormous expenses. There is a problem of becoming.

[0004]

SUMMARY OF THE INVENTION The present invention uses a living blood
A method for activating cellular immunity such as lymphocytes by heating to a temperature of 38 to 41 ° C. with a heat source of 5 to 80 ° C., a blood duct for taking out blood from a living body, and an intermediate of the blood duct, A heating mechanism for heating blood in the range of 55 to 80 ° C. and a blood delivery for adjusting the flow velocity of blood so that the heating temperature of blood is heated in the range of 38 to 41 ° C. according to the heating temperature of the heating mechanism. It is an object of the present invention to provide a device for activating cell-mediated immunity by heat treatment of blood, which has a simple structure of providing a pump, and solves such a problem.

[0005]

According to the method and apparatus of the present invention, blood is heated without causing denaturation or damage, and cellular immunity is activated.

[0006]

It is an object of the present invention to heat treat blood by using whole blood by selecting an appropriate temperature and circulation treatment time (unit blood flow rate), and increase the immune component of blood including simple treatment. Based on the mechanism, a method for activating cell-mediated immunity by heat treatment of blood, which leads to an effective treatment method with high safety against anti-tumor, anti-virus (including HIV), anti-infection and drug resistant diseases, and the like It is intended to provide an execution device.

[0007]

BACKGROUND OF THE INVENTION The present inventors have found that heat is applied to immunocompetent cells to remarkably enhance their immune activity.
As a result of conducting research on establishing an effective procedure of the heating method and elucidating the mechanism of action, it was confirmed that the method of heating blood effectively enhances cell-mediated immunity, particularly the activity of lymphocytes, He found that heating AIDS virus significantly suppressed the onset of AIDS virus.

There are two possible reasons for the suppression of the onset of AIDS. The first reason is that the AIDS virus is actively killed by the activation of lymphocytes, and the second reason is that the lymphocyte receptor for AIDS virus is heat-treated. It is to be inactivated.

It was also confirmed that heat-treatment of blood markedly activates cell-mediated immunity, and as a result of mixed culture of heat-treated lymphocytes and cancer cells, the cancer cells are significantly killed.

Further, in order to obtain immunological findings by heating blood, a flow type heating type device was devised, and circulation heating was performed using mouse spleen cells, rat blood and human blood as experimental materials. As a result of in vitro treatment (mixed culture) of target cells such as AIDS virus and mouse cancer cells, remarkable inhibition and killing action were observed.

[0011] Then, the mechanism of action thereof was examined, and the effect was not simply the expected heat-damaging effect due to heating, but the important role was the cell-mediated immunity in blood, particularly the lymphocyte activation enhancing effect. From the immunological point of view, he tried to activate the cellular immunity of blood and spleen cells for the purpose of treating diseases such as tumors and viral infections.

In the heat treatment, the effect of heat transfer to blood and spleen cells, which are experimental materials, is important.
The optimum conditions were examined by selecting (a) bottle type, (b) tube, and flow type. In the bottle-type heating experiment, heat transfer was not uniform, and heat damage to blood and spleen cells, especially large damage to lymphocytes, was observed. We thought that a stirring function would be required to increase the heat transfer efficiency, and the device would be complicated.

Therefore, a silicon tube having a high heat transfer effect,
I tried using flexible tubes such as Tygon tube, Viton tube, and Teflon tube. If the tube is thick, not only is the heat transfer efficiency poor, but there is a problem with the amount of blood introduced. On the other hand, when it is thin, the heat conduction efficiency is good, but the processing capacity decreases and it takes time. From these results, the outer diameter is 5.0 to 8.0 mm and the inner diameter is 2.5 to 4.0 mm.
It has been found that the flexible tube of No. 1 is suitable for use, and in particular, a silicon tube having an outer diameter of 6.4 mm and an inner diameter of 3.2 mm is suitable for obtaining and handling and is desirable. The length of the heating part depends on the thickness of the tube, and it is most important to find a condition that does not cause denaturation of cells due to heat injury. As a result of trial and error, a length of 200 to 600 mm is applicable. Yes, especially the outside diameter of 6.4 mm and the inside diameter of 3.2 mm
It was found that when using the above silicon tube, it is desirable that the length is about 400 mm. In addition, the use of this type has an advantage that the silicon tube is connected between blood vessels so that heating treatment can be performed while circulating blood. As a heating mechanism for the silicon tube, a constant temperature water bath or a metal block type constant temperature machine provided with a fitting groove for the tube can be used.

Since it is important to determine the temperature and flow rate (flow rate) of the heat source in order to avoid the alteration of blood and spleen cells and to raise a certain heating effect, the temperature and flow rate of the biomaterial (blood and spleen cells) are important. Was calculated.

In order to select a temperature for enhancing the cellular immune function such as blood components, it is necessary to first confirm that the biomaterial (spleen cells of mouse, blood of rat, human blood, etc.) is not denatured or damaged. Therefore, it can be used after pretreatment in a prescribed heating range, and then measuring the membrane resistance of erythrocytes at various salt concentrations and the plasma electrophoresis and enzyme activity (GOT, GPT, ALP, LAP, LDH) measurement. The safe temperature range was set at 55-80 ° C.

Further, when a blood vessel is directly taken out and circulated through a silicone tube, the site and thickness of the taken-out blood vessel may be affected by individual differences such as age, sex and weight, so the flow rate is from 30 ml / min. 100 ml / mi
The range was set to n.

As parameters of immune function, uptake of tritum thymidine ( ³ H-Thymidine), which is considered to be an index of immune function for lymphocytes, natural killer (nat).
The ural killer (NK) activity, interleukin-2 (IL-2) production, and killer subset / suppressor subset were measured, and the cases without heat treatment were compared for comparison. The experiment was carried out by using a silicon tube having an outer diameter of 6.4 mm and an inner diameter of 3.2 mm and setting the length of the heating portion to 400 mm.

Looking at the immune function index value after heat treatment, for example, the incorporation of tritum thymidine was compared with the control value (100% smoothed), the heating temperature was 70 ° C., and the flow rate was 100 ml / m.
The marked improvement was observed in general, such as remarkable activation showing a maximum of 465% in in., NK activity of up to 147% at a heating temperature of 55 ° C. and a flow rate of 40 ml / min. In addition, IL-2 productivity and k
Improvements in the iller subset / suppressor subset were also made by heat treatment (Table 1).

[0019]

[Table 1]

Using the results of these immune function indexes, the vertical axis represents temperature and the horizontal axis represents flow rate, and a graph is drawn that illustrates the safe / effective area, its boundary area, and the area of possible injury. We have made it possible to smoothly respond to fluctuations caused by (Fig. 1).

Further, the heating temperature of blood in this region was actually measured, and it was confirmed that the heating temperature at which excellent results were obtained was within about 38 to 41 ° C., and particularly the heating temperature near 39.5 ° C. was desirable.

[0022]

An embodiment will be described below with reference to the drawings.

FIG. 2 is a schematic view showing a first embodiment of the device of the present invention, in which a circulation type blood conduit 1 made of a silicon tube for connecting both ends to a vein of a living body H is formed, and the blood conduit 1 A blood delivery pump 2 for adjusting the flow rate of blood and a heating mechanism 3 are provided in the middle. A known means used in an artificial dialysis device, a blood transfusion device, or the like can be used for the connection part to the living body H, and a supply device 4 of an anticoagulant such as heparin is provided near the inlet of the blood duct 1. A supply device 5 of a neutralizing agent for neutralizing a blood coagulation preventing agent such as protamine is connected near the outlet, and a filter device 6 for separating coagulated blood is provided.

The flow rate of blood taken out from the living body H is controlled by the blood delivery pump 2, and the heating temperature of 55 to 80 ° C. is controlled in accordance with the flow rate of the blood passing through the heating mechanism 3. When the heating temperature is 55 to 80 ° C and the silicon tube constituting the blood conduit 1 has an outer diameter of 6.4 mm and an inner diameter of 3.2 mm, the flow rate is 30 to 100 (ml / min.) It is a safe adaptation value that does not cause denaturation, and the blood temperature in this range is 38 to 41 ° C, preferably 3
The flow rate and heating temperature are adjusted so that the temperature is around 9.5 ° C. The amount of blood to be heat-treated is about 1/3 of whole blood.
A sufficient effect can be expected with an amount of 1/4, and by doing so, the processing time can be set within a relatively short time of 10 to 30 minutes. The blood delivery pump 2 and the heating mechanism 3 may set and adjust the flow rate and the heating temperature respectively according to the calculation formulas, but either one is set as the main side and the sub side automatically sets and adjusts according to the setting of the main side. It can also be done.

FIG. 3 shows a second embodiment, in which a blood storage bag 7 is connected in the middle of the blood duct 1 after the heat treatment to temporarily store the blood having activated cell-mediated immunity and then It shows a device adapted to transfuse blood into a living body, and is otherwise the same as the first embodiment.

FIG. 4 shows a third embodiment of the present invention, in which the blood taken out from the living body H is stored in a blood storage bag 7 and heat-treated between blood lines 1 for transfusion to the original living body H. It is the same as the previous example except that an immune activation device was placed.

FIGS. 5 to 7 show the respective embodiments of the heating mechanism 3. First, FIG. 5 shows a flat plate body 9 having a fitting groove 8 into which the blood conduit 1 is fitted at a constant length on one surface thereof. Flat plate 9
The metal block type thermostat 13 is composed of a block body 11 having a lid body 10 for covering the fitting groove 8 side and a receiving tank portion 12 into which the block body 11 can be inserted and removed freely.

Next, FIG. 6 is provided with an inlet 14 and an outlet 15 for the blood conduit 1 on the upper surface, and the heated portion of the blood conduit 1 that has entered from the inlet 14 on the intermediate side circumference is wrapped around the outlet 15 at a constant length.
A metal block type thermostat 19 comprising a circular metal block 17 provided with a winding groove 16 for guiding to the inside and a receiving tank portion 18 into which the circular metal block 17 is freely inserted and withdrawn.
Water is filled in the voids in the receiving tank portion 18 into which the circular metal block 17 around which is wound is inserted so that the thermal conductivity is improved. Further, the winding groove 16 is formed in an inclined shape so that air does not stay inside when water is filled. Further, the depth of the receiving tank portion 18 is made deeper than the height of the circular metal block 17 in order to compensate for the loss due to the evaporation of water.

FIG. 7 shows an outer tank 20 and an inner tank 21 disposed inside the outer tank 20 for immersing the heated portion of the blood conduit 1 therein, and inside the outer tank 18 kept at a constant temperature by a control heater 22. The length of the blood conduit 1 for immersing the inner tank 20 in which the constant water level is kept constant by providing a pump 23 for injecting the constant temperature water into the inner tank 19 to cause the constant temperature water in the inner tank 20 to overflow. That is, the constant temperature water tank 24 that keeps the length of the heating part constant.
It shows. In addition to the overflow of the constant temperature water from the upper edge of the inner tank 20, a through hole 25 or a notched window 26 may be provided in the upper portion of the inner tank 20 as shown in FIGS. 8 (a) and 8 (b). .

The method for activating cell-mediated immunity by heat treatment of blood according to the present invention is carried out by using the above-described method and apparatus of the invention, and blood is heated at a heating temperature of 55 to 80 ° C.
The cell-mediated immunity is activated by heating to a temperature of 8 to 41 ° C.

Each experimental example of anti-AIDS virus and anti-cancer according to the present invention will be described below.

Anti-AIDS virus experiment AIDS is an immunodeficiency disease caused by AIDS virus, and it is necessary to consider the treatment from the viewpoint of cell-mediated immunity, and the effect of activating the immune function of lymphocytes by the heat treatment of blood of the present invention is a therapy. Can be expected as.

Generally, AIDS virus is vulnerable to heat, but this time, an experiment was carried out using the mouse AIDS virus (LP-BM5 MuLV), which is said to be close to human AIDS symptoms, under the above-mentioned heating conditions. . Preliminarily, when the mouse (B6) was inoculated with the mouse AIDS virus (LP-BM5 MuLV),
Hypertrophy of the spleen and lymph was observed as a specific symptom of viral infection. Next, in order to see the behavior of mouse AIDS virus against heat, the heating temperature was 70 ° C and the flow rate was 80 ml / mi.
n., 60 ml / min. and 40 ml / min.
After the virus suspension was heated, it was intraperitoneally administered to mice (B6). As a result, one of the infectious diseases, spleen enlargement, was observed, and no virus lethality was observed under these conditions.

Then, mouse splenocytes were taken out, heat-treated under the same conditions, and then mixed with untreated mouse AIDS virus (37 ° C., 6 hours, 5%, CO ² condition). After washing away the virus simply attached to the splenocytes, the splenocytes were intraperitoneally administered to mice (B6). As a result, no abnormality was observed compared with the spleen of a normal mouse. As a control group, spleen hypertrophy was observed when intraperitoneal administration of a mixture of splenocytes mixed with untreated mouse AIDS virus to a mouse (B6) was intraperitoneally administered without heat.

When the spleen is enlarged, not only the weight of the spleen is increased, but also the number of cells is increased as compared with the normal splenocytes. On the other hand, no change was observed in the spleen weight and the cell number in the mice in which the onset suppressing effect or the virus lethal effect was observed. This is because (1) the activity of splenocytes responsible for cell-mediated immunity (activity of lymphocytes) was increased, and the target AIDS virus was attacked and killed. Or (2) It is considered that it is due to the fact that the virus receptor on the splenocytes changes due to heat (inhibition of infection) (Table 2).

[0036]

[Table 2]

Anti-Cancer Experiment In order to examine the reaction of mouse cancer cells to heat treatment, the cells were cultured at 2 × 10 ⁶ cells / ml using mouse (ICR system) cancer cell Sarcoma 180 (S-180). Liquid (using RPMI 1640)
Prepared in, take 7 ml of each, and heat at 70 ℃
Set to 40ml, 60ml and 80ml /
Circulation heat treatment was performed through a silicon tube at a min. The control was untreated. After culturing, a fixed amount of cells was collected at 24 hours and 48 hours, stained with 0.2% trypan blue, and the number of stained dead cancer cells was measured to calculate the mortality rate.

Further, in order to observe the effect expected when heat treatment is applied to splenocytes involved in the cell-mediated immune function,
The spleen of a mouse (ICR) is taken, separated into single cells with a culture solution (using RPMI 1640), and heat-treated under the above-mentioned conditions. Then, using the same mouse cancer cells (untreated), spleen cells and cancer cells are separated. A mixed culture was carried out at a ratio of 30: 1 at a temperature of 37 ° C. for 48 hours, and staining treatment was performed in the same manner to determine the mortality of cancer cells.

As a result, no difference was observed in the heat-treated group of cancer cells at 70 ° C./80 ml as compared with the control, but at 70 ° C.
Under the more severe conditions of / 60 ml and 70 ° C / 40 ml, it was 18,48% and 22.5% in 24 hours. Further, at 48 hours, cell death increased as the conditions became more severe. On the contrary, in the mixed culture of heat-treated spleen cells and cancer cells, the mortality rate of 12.9% of the control was 2% at 70 ° C / 80 ml.
9.31%, increased to 38.4% at 70 ° C / 60 ml,
The mortality rate decreased to 20.46% at 70 ° C / 40 ml. Further culturing further increased mortality, compared to 22.93% of the control, increased to 57.23% at 70 ° C / 80ml and 78.78% at 70 ° C / 60ml, but more severe. Mortality decreased to 32.15% at 70 ° C / 40 ml.

A significant difference in the death rate of cancer cells was observed between the heat-treated group of cancer cells and the heat-treated group of spleen cells, because the activation of splenocytes by heat stimulation killed the cancer cells. is there. On the other hand, the severe conditions are 70 ℃ / 40ml
Therefore, it is considered that the heat damage to the spleen cells themselves was great. In the circulating heat treatment of cancer cells alone, a slight increase in the lethality was observed by extending the heating time and slowing the flow rate.

This strongly supports that heat treatment of blood, splenocytes and the like activates the cell-mediated immune function and, as described above, can be a useful means for hyperthermia treatment of cancer. (Table 3).

[0042]

[Table 3]

[0043]

INDUSTRIAL APPLICABILITY As described above, the present invention can activate cellular immunity such as lymphocytes by heat-treating blood at a heating temperature in a range that does not cause cell degeneration or damage. By enhancing the immunity, there is an effect that it can be expected as an effective treatment for infectious diseases and drug resistant diseases.

By adopting the circulation heating method, there is an effect that the heat treatment time can be shortened.

By adopting the blood storage method, it is possible to obtain a time margin, and it is possible to easily confirm the presence or absence of degeneration of cells.

Since it can be carried out by a simple device such as a flexible tube such as a silicon tube which is easily available and convenient to handle and a heating temperature and flow rate controlling means, the apparatus can be provided at a low price. Moreover, since it can be widely spread, there is an effect that the physical, time, and economic burden on the user can be reduced.

When the heating mechanism is a metal block type constant temperature machine, the flexible tube is fitted into the fitting groove of the block body,
Alternatively, since the preparation can be performed by a simple work of winding the winding groove, there is an effect that the handling can be facilitated and the time can be shortened.

Further, by filling the space in the receiving tank portion containing the heated portion of the flexible tube with a liquid, the heat conduction to the blood can be made uniform and the heat treatment can be performed more precisely.

When the heating mechanism is a double-structured constant temperature water tank, the liquid in the inner tank for accommodating the flexible tube is made to overflow, so that the liquid level in the inner tank is not affected by evaporation of the liquid. The effect is that the heating temperature of the blood can be accurately maintained without changing the length of the heating portion of the flexible tube.

[Brief description of the drawings]

FIG. 1 is a graph showing a safe / effective area depending on a heating temperature and a flow rate of blood.

FIG. 2 is a schematic diagram showing a first embodiment of the present invention.

FIG. 3 is a schematic diagram showing a second embodiment of the same.

FIG. 4 is a schematic view showing the same as the third embodiment.

FIG. 5 is a perspective view showing a metal block type constant temperature machine which is the first embodiment of the heating mechanism in a state before being set.

6A and 6B show a second embodiment of the present invention, in which FIG. 6A is a perspective view showing a state before setting, and FIG. 6B is a vertical sectional view showing a use state after setting.

FIG. 7 is a vertical sectional view showing a constant temperature water tank of the third embodiment.

FIG. 8 is a partial perspective view showing another two examples of the inner tank.

[Explanation of symbols]

 1 is a blood conduit (flexible tube) 2 is a blood delivery pump 3 is a heating mechanism 4 is an anticoagulant supply device 5 is a neutralizer supply device 6 is a filter device 7 is a blood storage bag 8 is a fitting groove 9 Is a flat plate body 10 is a lid body 11 is a block body 12 is a tank section 13 is a metal block type thermostat 14 is an inlet 15 is an outlet 16 is a winding groove 17 is a circular metal block 18 is a receiving section 19 is a metal block type thermostat 20 Is an outer tank 21 is an inner tank 22 is a controlled heater 23 is a pump 24 is a constant temperature water tank 25 is a through hole 26 is a notched window

Front Page Continuation (72) Inventor Kenichi Takayama 561-104 Katakura-cho, Hachioji City, Tokyo, Takayama Institute of Life Science Co., Ltd. (72) Inventor Takashi Katsura, 2693 Nishikata, Koshigaya, Saitama Prefecture

Claims (14)

[Claims] 1. A cell by heat treatment of blood, characterized by activating cell-mediated immunity such as lymphocytes by heating living blood to a temperature of 38 to 41 ° C. with a heat source of 55 to 80 ° C. Method of activating sexual immunity. 2. The method for activating cell-mediated immunity by heat treatment of blood according to claim 1, wherein the blood is taken out of the living body, heat-treated outside, and circulated to the original living body. 3. The method according to claim 1, wherein blood is taken out from the living body, heat-treated, and once stored, the blood is then transfused into the original living body.
The method for activating cell-mediated immunity by heat treatment of blood according to 1. 4. Taking blood from a living body and temporarily storing the blood,
The heat treatment is performed when the blood is transfused into the original living body.
A method for activating cell-mediated immunity by heat treatment of blood according to the description. 5. A blood conduit for extracting blood from a living body, a heating mechanism for heating blood in the range of 55 to 80 ° C. in the middle of the blood conduit, and a blood temperature corresponding to the heating temperature of the heating mechanism. A device for activating cell-mediated immunity by heat treatment of blood, which is provided with a blood delivery pump for adjusting the flow rate of blood so that the heating temperature is heated in the range of 38 to 41 ° C. 6. The apparatus for activating cell-mediated immunity by heat treatment of blood according to claim 5, wherein the blood duct is a circulation duct that circulates to a living body. 7. The apparatus for activating cellular immunity by heat treatment of blood according to claim 5, wherein a blood storage bag is provided at the end of the blood duct. 8. A heating means for heating the blood in the range of 55 to 80 ° C. in the middle of the blood conduit for transfusing the blood in the blood storage bag to the original body, and the blood corresponding to the heating temperature of the heating means. A device for activating cell-mediated immunity by heat treatment of blood, which is provided with a blood delivery pump for adjusting the flow rate of blood so that the heating temperature is heated in the range of 38 to 41 ° C. 9. The blood conduit has an outer diameter of 5.0 to 8.0 mm and an inner diameter of 2.5.
Flexible tube such as silicone tube, Tygon tube, Viton tube, Teflon tube, etc., with a length of 200m to be heated by the heating mechanism
The apparatus for activating cell-mediated immunity by heat treatment of blood according to any one of claims 5 to 8, which has a size of m to 600 mm. 10. A block body having a flat plate portion in which a heating mechanism has a flexible tube fitting groove formed on one surface, a lid body covering the fitting groove of the flat plate portion, and a receiving tank into which the block body is freely inserted and removed. The activation device for cell-mediated immunity by heat treatment of blood according to any one of claims 5 to 9, which is a metal block type thermostat consisting of a part. 11. A heating mechanism is provided with an inlet and an outlet of a flexible tube on an upper surface thereof, and a winding groove for winding a heating portion of the flexible tube that has entered from the inlet and guiding it to the outlet on an intermediate side circumference. The apparatus for activating cell-mediated immunity by heat treatment of blood according to any one of claims 5 to 9, which is a metal block thermostat consisting of a circular block provided and a receiving section into which the circular block can be inserted and removed freely. 12. The apparatus for activating cell-mediated immunity by heat treatment of blood according to claim 11, wherein a liquid is filled in a void in a receiving tank portion into which a circular block around which the flexible tube is wound is fitted. 13. The apparatus for activating cell-mediated immunity by heat treatment of blood according to claim 12, wherein the winding groove is formed in an inclined shape so as to prevent air in the receiving tank portion from staying when the liquid is filled. 14. A pump, wherein the heating mechanism comprises an outer tank and an inner tank which is disposed in the outer tank to immerse a heated portion of the flexible tube, and which pumps the liquid in the outer tank into the inner tank. 10. A constant temperature water tank in which a liquid level in the inner tank is maintained by maintaining a liquid surface height of the inner tank by maintaining the immersion length of the flexible tube constant by providing a liquid. An apparatus for activating cell-mediated immunity by heat treatment of blood according to any one of 1.