JP3354669B2 - Biological reaction test method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an interleukin 1 (hereinafter referred to as IL-
The present invention relates to a method for examining a biological reaction relating to immunity, inflammation and the like by inducing the production of 1) and measuring the production ability of the IL-1.

[0002]

2. Description of the Related Art As well known, as immune cells responsible for the immune monitoring mechanism of living organisms, leukocytes such as monocytes, macrophages, granulocytes or lymphocytes, killer cells, NK cells or LAK cells play an important role. Play. These cells perform immune monitoring in the blood and in various organs and organs, and play various roles in various immune reactions.

[0003] It is generally known that the functions of these cells are suppressed or enhanced by various immune reactions in various diseases and malignant tumors. Therefore, by examining the functions of these cells, it is possible to understand the immune function of the living body. For example, in immunological tests for various diseases, tests for immunocompetent cells
Lymphocyte blastogenesis test, neutrophil phagocytic function test, neutrophil bactericidal ability (active oxygen production ability)
Tests are being conducted.

On the other hand, various immune factors and the like secreted from these immune cells also play an important role in the body's immune system. In particular, due to the recent development of genetic engineering technology and the like, the gene structures of many humoral immune factors (such as cytokines) have been elucidated using recombinant DNA technology. A variety of cytokines such as interleukins, interferons, and TNF have been reported so far, and these factors are closely related in vivo and are very greatly involved in the immune function of the living body. I understand. At present, it is considered that knowing the dynamics of these cytokines and the like is extremely important in considering the immune system of a living body.

[0005] Therefore, in order to understand the immunological function of the living body against various diseases and malignant tumors, the behavior of these cytokines in the blood of patients has recently been investigated. However, since these cytokines act in vivo in a very small amount, it is very difficult to grasp the behavior in vivo. It is also important to measure the blood levels of these cytokines, but it is also important to go one step further and know the ability of blood to produce or react with these factors.

As a method for examining the immune function of a living body, in addition to grasping the behavior of cytokines in the living body, the function of the immunocompetent cell itself derived from the patient is examined. For example,
Interferon-γ-producing ability and interleukin-2-producing ability of leukocytes of patients have been examined as immunological tests. By examining the ability of the cells themselves to produce cytokines, it is also possible to grasp biological reactions.

[0007] IL-1 is a cytokine mainly produced from cells such as monocyte / macrophage cells and neutrophils. IL-1 acts on various cells, exhibits various biological activities, and plays an important role in biological reactions such as immunity, inflammation, hematopoiesis, endocrine and cranial nerves (Oppenheim, JJ et.
al .: There is more than one interleukin 1.Immunol.
Today, 7, 45-56, 1986).

[0008] Further, IL-1 is constantly produced even in a normal solid, and is considered to be an essential factor for maintaining the function of a living body. On the other hand, it has also been suggested that when IL-1 is abnormally produced, it may cause a disease accompanied by inflammation. For example, in a cerebrospinal fluid of a patient with rheumatoid arthritis,
Since L-1 activity has been detected and a correlation has been observed between the activity and the degree of progress, it has been suggested that IL-1 is involved in the disease state (Eastgate, JA et al .: Cor
relation of plasma interleukin 1 levels with disea
se activity in rheumatoid arthriitis, Lancet, 891
3, 706-709, 1988). In addition, IL-1 has been suggested to be involved in various diseases such as glomerulonephritis, granuloma, Alzheimer's disease, Down's syndrome, Kawasaki disease, gout, and acute myeloid leukemia. IL-1 has a wide variety of actions in various tissues and cells throughout the body, and is involved in various responses of the living body to diseases such as infectious diseases and inflammatory diseases. Thus, it has become extremely important to know the production, production ability, and dynamics of IL-1 during a disease state.

[0009] However, there are not many reports that clearly show the relationship between the amount of IL-1 produced in blood and diseases. The reason for this is that since the action of IL-1 is very strong and acts at a concentration of pg / ml, changes in the amount of production that can cause functional abnormalities in living organisms cannot be accurately detected by current measurement systems. There is a possibility.

Therefore, recently, many studies have been conducted to collect peripheral blood monocytes and macrophages, examine their IL-1 producing ability in vitro, and clarify the relationship between disease and IL-1 producing ability. Have been done. However, collection and preparation of monocytes and the like from blood requires complicated operations including aseptic treatment, and the procedure during the operation greatly affects cells.

When lipopolysaccharide (LPS) or various drugs are used as stimulants, their IL-1 producing ability is greatly affected by the reactivity of a specific stimulant such as its receptor. That is, the I of a certain individual
It is considered that the sensitivity of a cell receptor to a certain drug has a greater effect than the L-1 production ability.
It is difficult to grasp the L-1 production ability. In addition, all of these tests are performed in a system in which various humoral factors and other cells contained in blood are not present, and cannot be said to accurately reflect a biological reaction in the body.

[0012]

SUMMARY OF THE INVENTION The object of the present invention is different from the conventional measurement of IL-1 producing ability by stimulants such as LPS and various drugs, and is simplified by using a blood sample such as whole blood. It is an object of the present invention to provide a novel biological reaction test method capable of measuring IL-1 production ability with a measurement system.

[0013]

The present invention SUMMARY OF], the center line average roughness Ra Ri 20μm der from 0.2 [mu] m, bumpy average
By spacing Sm value contacting the material with a blood sample with the following der Ru surface roughness 200 [mu] m, to induce the production of IL-1, biological, characterized by measuring the production amount of the IL-1 This is a reaction test method.

In the biological reaction test method of the present inventor, whole blood or its diluted blood or blood cells can be used as the blood sample, and whole blood or the like can be used as it is. There is no need to separate peripheral monocytes, neutrophils, etc. from the blood, and of course the operation and processing time involved are not required, as well as those operations,
There is no problem that the activity of the cell function is reduced or unnecessary activation is caused by the treatment time or the like. Furthermore, in the present invention, the amount of blood and the like used can be very small, and the burden on the blood-collecting person can be significantly reduced. In addition, since whole blood or its diluted blood is used, the involvement of various humoral factors and cells contained in the blood works in the same manner as in the living body, and the in vivo protective function can be grasped more accurately.

Hereinafter, the present invention will be described in detail. In one example of the embodiment of the method of the present invention, blood is collected as a sample, and this blood is added to a plate well, a test tube or the like as whole blood or diluted. These plate wells and the bottom or side surfaces of the test tube inner wall have IL-
1 In order to induce the production, a surface roughness in the above specific range is imparted, or a material having the surface roughness is filled. By culturing blood using these containers, the material having the surface roughness and the blood act to induce IL-1 production. By measuring the amount of IL-1 produced, biological reactions related to immunity, inflammation and the like can be grasped.

In the above, collection of a blood sample can be performed by an arbitrary method according to a conventional method. For example, heparin blood sampling, citrate blood sampling, and the like can be mentioned. For dilution, for example, phosphate buffer, Hanks' solution, MEM, RPM
Any ordinary medium such as I-1640 can be used. Culture is usually performed under a temperature condition of about 37 ° C.
C. to 42.degree. C. can also be performed.

It is preferable to mix a plate or a test tube with a shaker or a rotary incubator at the time of culturing to induce the production of IL-1, and it is easy to grasp the production ability of each blood sample. The measurement of the amount of IL-1 produced after the culture can be performed by various immunoassays, typically an immunoenzymatic antibody method. In addition, since this method for inducing IL-1 production does not use LPS, lectin, or the like, when the IL-1 activity is evaluated by a cytotoxicity test, there is no effect of these stimulants on various cells, and accurate cytotoxicity is obtained. Sex can be measured. Thus, the production of IL-1 can be accurately measured using various cytotoxicity tests.

Next, the surface roughness given to the material to be brought into contact with the blood sample will be described. It is known that cells such as leukocytes in blood phagocytose foreign substances and are sticky cells. In the living body, biological defense is performed by poorly invading invading bacteria and the like, but leukocytes also poorly eat fine particles made of a polymer such as latex. Further, when a polymer material is embedded in a living body or when blood and the polymer material are brought into contact with each other, adhesion of leukocytes and platelets occurs. Various reactions such as release of intracellular enzymes and release of activated oxygen occur when these phagocytosis and adhesion effects are exerted. These reactions are also affected by properties such as surface properties and composition of the polymer material to be deprived or adhered.

Hitherto, regarding the phagocytosis and adhesion of cells, the particle diameter, fiber diameter and composition of the material have been investigated, but the shape of the material surface has not been well understood. Therefore, we have studied the interaction between cells and the surface shape of polymer materials. As a result, it was found that the roughness or unevenness of the material surface caused the production of IL-1 by leukocytes, and by controlling the shape of the material surface, the IL-
Studies have been conducted to induce the production of Escherichia coli 1 with high efficiency. Then, in order to induce the production of IL-1, the value of the center line average roughness Ra is 0.2 μm to 20 μm, preferably,
0.2~10μm der is, and bumpy mean spacing Sm value is found that is required to have the uneven surface is 200μm or less.

We have also found that even under induction under the same conditions, the amount of IL-1 produced varies from individual to individual. It was recognized that this action was extremely useful for preliminarily grasping the potential bioreactivity of an individual, and reached the present invention. Exogenous microorganisms, fungi, cancers, and the like are considered to be a kind of foreign matter to the living body (host), and these materials are also foreign matters to the living body. It is thought to reflect the function well.

[0021] Further, it has been revealed that IL-1 plays an important role also in immunity and inflammatory response. Measuring IL-1 production ability using the blood of a healthy person serves as an index for understanding the individual's immunity and inflammation-related bioreactivity. It can also be used as a progress marker for disease.

The above Ra value is defined in JIS B0601-19.
It is the center line average roughness at 82. Further, the value of the irregular average interval Sm is a value defined as follows.

The average roughness Sm value of the irregularity The current JIS standard specifies the information of the surface roughness in the height direction, but does not specify the information of the surface direction. However, the irregularities in the present invention are:
Preferably, it is also limited by the spacing of the irregularities in the plane direction, as is apparent from the examples described later. Therefore,
In the present invention, the range of the unevenness in the plane direction is defined by preferably using the average mean spacing Sm value.

The value of the irregular average interval Sm is obtained as follows. First, an upper count level and a lower count level are drawn at positions of a constant height and depth, respectively, with respect to the center line B of the roughness curve A shown in FIG.
Next, when a point where the upper count level intersects with the roughness curve at least once intersects between two points where the lower count level intersects with the roughness curve A, a “mountain” is defined as one mountain. Is defined.

The average value of the average interval Sm is shown in FIG.
As shown in the figure, the interval between the peaks between the reference lengths L is Smi.
Is a value defined by the following equation.

[0026]

(Equation 1)

That is, the value of the average mean interval Sm is as follows:
The average value of the interval between the peaks located between the reference lengths L is shown. In this manner, the condition of the surface direction of the unevenness is defined by the average interval Sm value of the unevenness.

The present inventors conducted an IL-1 production induction test using various films, plates, porous particles, and the like having different Ra values. As a result, the center line average roughness Ra value was 0.1%.
When a material such as a film, a plate, or a porous particle having a surface roughness of 2 μm to 10 μm is brought into contact with blood or the like,
It was found that the induction of IL-1 production was significantly enhanced.

Therefore, in order to enhance the induction of IL-1 production, the center line average roughness Ra value needs to be 0.2 μm to 20 μm. In addition, the present inventors have found that R
By polishing a PET film having an a value of 0.2 μm or more and 20 μm or less with a 1200-mesh sandpaper while changing the polishing time, the Sm value was 474, 374, 2 or more.
Each film of 13, 101, 56, 31 μm is made,
As a result of an IL-1 production induction test using a film described below, it was found that when the Sm value was 200 μm or less, the induction of IL-1 production was significantly increased.

As a material that can be used in the present invention, any material can be used as long as no harmful metals or additives such as plasticizers are eluted when the material comes into contact with blood or the like. Can be. That is, synthetic and natural organic polymer materials and inorganic materials such as glass and alumina can be used, and the surface has a center line average roughness Ra value of 0.2 μm to 2 μm.
Any material may be used as long as the material has irregularities of 0 μm. Synthetic and natural organic polymer materials include cellulose acetate, polystyrene, nylon, polytetrafluoroethylene, polytrifluoroethylene, perfluoroethylene-propylene copolymer, polyethylene terephthalate, polyethylene, polyvinyl chloride, acrylic resin,
Ethyl cellulose and the like can be exemplified.

As a method for imparting surface roughness, polishing is generally used, but other methods can also be used to prepare a material having the above specific surface roughness. For example, a polymer porous material obtained by a method of physically or chemically fixing fine particles on the material surface, a suspension polymerization method, or a material having a porous surface obtained by a sol-gel method can be used.

As a method for fixing the fine particles on the surface of the material, for example, a method of coating fine particles of 0.1 μm to 20 μm can be mentioned, whereby the production of IL-1 can be induced. The fine particles used here can be prepared, for example, by emulsion polymerization or suspension polymerization of a styrene-based or acrylic-based vinyl monomer alone or a mixture of two or more components. Desirably, these fine particles are made of a copolymer with a polyfunctional monomer having two or more functional groups such as divinylbenzene.

Next, the fine particle coating method will be described. First, 0.1 to 5% by weight of a synthetic polymer or a natural polymer as a binder for coating these fine particles.
Suspend in the solution in which it has been dissolved to a certain extent. Next, a bead, fiber or film-like material made of an organic or inorganic material previously formed is immersed in the suspension, and then dried to produce the IL-1 production-inducing material of the present invention. Can be. In this case, the Ra value can be adjusted by the size of the fine particles, and the Sm value can be adjusted by the concentration of the suspended particles.

Also, in the case of a porous material, the shape may be any of a film shape, a fiber shape and a bead shape, and only the surface may be porous, or the whole material may be porous. . For the surface acoustic wave porous material, for example, a synthetic polymer or a natural polymer as a coating solution is added to a bead, fiber or film-like material made of an organic or inorganic material in a good solvent thereof in an amount of about 0.1 to 5% by weight. It is obtained by spraying the dissolved liquid into a mist by spraying and drying by heating.

In addition, as a method for manufacturing the whole material depending on the porosity, a general method for manufacturing a porous membrane, a porous bead or the like can be used. For example, in the case of a film, the synthetic polymer is dissolved in a good solvent, cast on a glass plate, and then the film is washed with a poor solvent of the synthetic polymer, and the good solvent is extracted. A membrane can be prepared.

The adjustment of the Ra value and the Sm value in the case of a porous material is achieved by adjusting the pore diameter and the pore amount.
The pore diameter and pore amount can be easily adjusted by changing the type and amount of the solvent in which the material is dissolved. In the case of particles or the like produced by the suspension polymerization method, it can be adjusted by changing the amount of a high boiling point poor solvent that does not dissolve in or dissolves in the polymer or the suspension phase.

The temperature at which the material is brought into contact with the blood sample ranges from 15 ° C. to 4 ° C. in consideration of the effect on cells and proteins.
2 ° C is desirable, but 25 ° C rather than the range of 15 ° C to 25 ° C
The range of -42 ° C is preferable because the induction of IL-1 production is greater.

[0038] For these reasons, using a material having Ri center line average roughness Ra value 0.2μm~20μm der and bumpy mean spacing Sm value of unevenness in the following range 200μm on the surface, such as blood Can induce IL-1 production, and can be used for an immune function test.

[0039]

In the present invention, the mechanism by which IL-1 production is induced when a material having a specific surface shape comes into contact with blood or blood cells is not always clear, but these materials interact with cells. It is considered that the production of IL-1 is induced by performing the treatment. IL-1
The cells that produce are not limited to cells in peripheral blood, but also include cells obtained from lymph vessels, lymph nodes, spleen, and the like. The blood is rich in these cells that produce IL-1. It is also considered that even if these IL-1 producing cells in the blood do not directly act on the material, some factor acts on the material, and IL-1 production is induced by another induced factor. Can be

[0040]

Next, the present invention will be described in more detail by giving Examples and Comparative Examples of the present invention.

First, the following Examples 1 to 17 and Comparative Example 1
The method for preparing the sample and the test method used in Nos. 13 to 13 will be described. (1) Method for Producing Polished Film (Examples 1 to 9, Comparative Examples 1 to 9) After preparing a film as an IL-1 production inducing material and washing one surface of the film with methyl alcohol,
Struers (Denmark), automatic polishing machine (trade name:
Plano Ball Pedemax), 220, 500, 12
Polishing was performed using a sheet having 00, 2400, and 4000 mesh side papers attached thereto to produce a polishing film having a surface roughness on one side of the surface.

(2) IL-1 Production Induction Test Method Using Polished Film (Examples 1 to 9 and Comparative Examples 1 to 9) The film was punched into a 15 mm diameter, and the surface with the above-mentioned surface roughness was turned upward. Well plate (Nun
c, multi-dish, manufactured by InterMed). After washing these wells with physiological saline for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.), 2.0 ml of heparin-collected healthy human fresh blood was added to the wells to which the film was adhered, and placed on a shaker. The cells were cultured with shaking for an hour.

Thereafter, blood was collected and the concentration of IL-1 in plasma was measured by the method described below. (3) Experiment of Inducing Tumor Necrosis Factor Production by Attachment of Microparticles (Examples 10 to 17 and Comparative Examples 10 to 14) Each microparticle was placed on the bottom of a well of a 24-well plate (Nunc, Multidush, manufactured by InterMed) as described below. And attached. After washing these wells with physiological saline for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.), 2.0 ml of heparin-collected healthy human fresh blood was added to the wells, placed on a shaker, and shaken at 37 ° C. for 2 hours. While culturing.

Thereafter, blood was collected, and the concentration of IL-1 in plasma was measured by the method described below. (4) Surface roughness measuring method The center line average roughness Ra value and the uneven average spacing Sm value described in each of Examples and Comparative Examples are measured by a surface roughness measuring device (manufactured by Kosaka Laboratory Co., Ltd., device name). : Surf Coder S
E-30D) or tomographic electron microscope (ELIONIX)
(Manufactured by the company, device name: ESA-3000).

(5) Method of Measuring IL-1 Concentration in Plasma Blood after the IL-1 production induction test was centrifuged to collect plasma, and the concentration of IL-1β in the plasma was determined using an IL-1β monoclonal antibody. , Immunoenzyme antibody method (R & D Syst
product name: Quantikine IL-1
β).

The detection limit concentration of this measuring method is 7 pg / m.
l. In addition, the blood immediately after blood collection was centrifuged to collect plasma, and the concentration of IL-1β in the plasma was measured in the same manner. The plasma IL-1β concentration of the blood immediately after the blood collection was below the detection limit.

Examples 1 to 3 and Comparative Examples 1 to 3 Using a PET film (polyethylene terephthalate film, manufactured by Unitika Ltd., trade name: EMBLET S-75), polishing was performed in five steps according to the above-described method for producing a polishing film. The prepared film was produced. Then, the Ra value was determined for each of the unpolished film and the film polished in five steps, and then the IL by the above-described film was determined.
-1 production induction test was performed. The results are shown in Table 1 below.

[0048]

[Table 1]

As is clear from Table 1, the Ra value was 0.6.
Above 1 μm, induction of IL-1 production increased rapidly.

Examples 4 to 6 and Comparative Examples 4 to 6 A PET film was prepared from a CA film (manufactured by Art Plus, cellulose acetate film, trade name: acetyl film VR-).
R), and instead of washing with methyl alcohol, soxhlet extraction (24 hours) was performed with methyl alcohol to extract the plasticizer, and the film was taken out.
After air-drying for hours, an unpolished and polished film was prepared in the same manner as in Examples 1 to 3 and Comparative Examples 1 to 3, except that the film was further dried at 80 ° C for 5 hours, and an IL-1 production induction test was performed. . Table 2 shows the results.

[0051]

[Table 2]

As is clear from Table 2, a polishing film (Ra) polished with a sandpaper of 1200 mesh or less.
(Value = 0.58 μm or more), the production induction of IL-1 increased rapidly.

Examples 7 to 9 and Comparative Examples 7 to 9 Examples 1 to 3 except that polishing was performed using a 500-mesh sandpaper and the polishing time was changed.
The surface roughness Ra value is 1.0 μm or more and
Five types of polishing films having values in the range of 20 to 370 μm were produced (see Table 4). The above five types of polishing films and an unpolished PET film (Comparative Example 7) were prepared, and an IL-1 production induction test was performed in the same manner as in Example 1.
Table 3 shows the results.

[0054]

[Table 3]

As is apparent from Table 3, the Sm value was 200
The Sm value was 20 compared to Comparative Examples 7 to 9, which exceeded μm.
In Examples 7 to 9 in which the thickness is 0 μm or less, it is found that induction of IL-1 production is remarkably large. That is, the same level of R
It can be seen that the effect of inducing IL-1 production is further enhanced when the Sm value is 200 μm or less even if it has an a value.

Example 10 and Comparative Example 10 A 24-well plate was mixed with styrene-divinylbenzene copolymer (copolymerization ratio: 1: 1) fine particles (manufactured by Sekisui Chemical Co., Ltd., particle size: 3 μm) prepared by suspension polymerization. Cellulose acetate suspended by weight% (manufactured by Daicel Chemical Industries, Ltd., trade name:
Linter) was immersed in a 1% by weight solution of methyl chloride-ethanol (9: 1 by weight) and then air-dried, thereby producing a fine particle coated plate (Example 10).

It was confirmed by an electron microscope that the fine particles were coated with cellulose acetate on the bead surface. The fine particle coated plate obtained as described above (Example 10) and a plate immersed in a similar solution containing no fine particles (Comparative Example 10)
Was used to conduct the above-mentioned IL-1 production induction test. Table 4 shows the results.

[0058]

[Table 4]

As apparent from Table 4, the Ra value of the fine particle coated plate (Example 10) was 2.46 μm.
m, and the induction of IL-1 production was 30 times higher than that of Comparative Example 10.

Example 11 and Comparative Example 11 A 24-well plate was coated with a 5% by weight solution of cellulose acetate resin (manufactured by Daicel Chemical Industries, Ltd.) in methylene chloride / methanol (9: 1 by weight) as a coating solution. (Freund Sangyo, product name: FL0-
5) was sprayed to prepare a plate coated with a cellulose acetate resin. The coated plate was photographed with a scanning electron microscope and observed. As a result, it was confirmed that a porous bottom surface having a 200 μm-thick coating layer was obtained. The coating conditions are as follows.

Spray air pressure: 3.5 kg / cm ² Spray liquid temperature: room temperature Spray liquid flow rate: 100 ml / min Spray temperature: 60 ° C. Spray nozzle diameter: 1.2 mm Coating plate obtained as described above (Example 1)
An IL-1 production induction test was performed using 1) and an uncoated plate (Comparative Example 11). Table 5 shows the results.

[0062]

[Table 5]

As is clear from Table 5, the Ra value of the coated plate (Example 11) was 1.18 μm, which was lower than that of the uncoated plate (Comparative Example 11).
Induction of IL-1 production showed a 20-fold performance.

Examples 12, 13, 14, 15 and Comparative Example
12 and 13 of the particle diameter 0.20μm polystyrene latex particles (Polybead: Polysciences, In
c. 500 μl of 50,000 particles / ml and 1 ml of acetone were simultaneously added to a 24-well plate, and the latex particles were welded to the bottom of the well. After 10 seconds, 1 ml of distilled water was added to stop the welding. After washing with distilled water and methanol, it was air-dried and washed with physiological saline for injection. Similarly, particle diameters 0.5, 1.0, 6.0, 20.
0,45.0 μm particles were deposited.

Using these latex-welded plates, the above-mentioned IL-1 production induction test was performed. Table 6 shows the results.

[0066]

[Table 6]

As is apparent from Table 6, the Ra value was 0.0
In a plate from 4 μm or more to 9.63 μm or less,
As the Ra value increased, the induction of IL-1 production increased rapidly.

Examples 16, 17, 18 and Comparative Example 14 Porous polystyrene particles Diaion HP-50, HP
-20 (manufactured by Mitsubishi Kasei), polyacrylester porous particles Diaion HP-1MG (manufactured by Mitsubishi Kasei), polyacrylester porous particles Amberlite XAD-7
(Manufactured by Organo Corporation) was washed three times with distilled water and allowed to stand at room temperature overnight. After washing with distilled water three times, washing with methanol, the resultant was suspended in methanol and allowed to stand at room temperature for 6 hours. This was washed with distilled water and physiological saline for injection, suspended in saline for injection, subjected to ultrasonic cleaning, and degassed using a suction pump.

Each of the particles thus washed was filled into a sterilized and washed polypropylene tube (for Eppendorf Safelock Tube 2.0 ml, manufactured by Diatron Co., Ltd.) in a volume of 500 μl. 1.6 ml of blood collected from heparin from a healthy person was added to this tube,
The mixture was gently inverted at 37 ° C. for 2 hours. The blood was taken out, the plasma was separated, and the plasma IL-1 concentration was measured by the above-described IL-1 concentration measurement method. Table 7 shows the results.

[0070]

[Table 7]

As is clear from Table 7, the effect of inducing TNF is not seen with HP-20 having a surface roughness Ra smaller than 0.2 μm, but HP-20 having a surface roughness Ra larger than 0.2 μm.
50, remarkable induction of IL-1 production was observed in HP-1MG and XAD-7.

[0072]

According to the present invention, it is only necessary to prepare a material having a surface roughness in the above-mentioned specific range and contact it with a blood sample, so that IL-1 can be easily and rapidly produced from blood. Can be guided. If this is used, for example, a disease state can be grasped by monitoring a change in IL-1 production ability at various disease states. Further, even in a healthy person, the ability to produce IL-1 can be monitored to prevent various diseases. Alternatively, since the ability of cells to produce IL-1 can be checked in advance outside the body, it can be a marker for determining the drug dose and treatment method for cancer and various diseases. In this way, biological reactions related to immunity, inflammation and the like can be grasped.

[Brief description of the drawings]

FIG. 1 is a view for explaining the definition of a peak in an uneven average interval Sm value.

FIG. 2 is a diagram for explaining an uneven average interval Sm value.

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-5-168706 (JP, A) JP-A-6-319796 (JP, A) JP-A-6-2099992 (JP, A) 98599 (JP, U) (58) Field surveyed (Int. Cl. ⁷ , DB name) G01N 33/50-33/98

Claims (1)

(57) [Claims] 1. A center line average roughness Ra of 0.2 μm to 2 μm.
0μm der is, bumpy average interval Sm value is 200μm or more
By contacting the material with a blood sample with a lower der Ru surface roughness, to induce the production of interleukin-1, biological response test method characterized by measuring the production amount of the interleukin-1.