JPH11267199A - Blood treatment method and device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To remove or inactivate a target substance such as a hazardous substance in blood by returning blood to a body after processing the blood by a carrier having a compound that adsorbs, removes or inactivates at least one target substance of hazardous substances in blood taken from the body and an anticoagulant on the surface. SOLUTION: Blood drawn by a drawing means having a drawing needle 8, after blood corpuscles are processed with a blood corpuscle processing device 6, is separated into plasmas and corpuscles with a blood plasma separator 4, and the plasmas are processed with a plasma processing device 5. As the plasma separator 4, a hollow thread type plasma separator or a centrifugal plasma separator can be cited. The plasma processing device 6 is filled with a carrier such as a porous flat membrane, hollow thread membrane, nonwoven fabric, woven fabric, or a porous bead in the housing. A compound having compatibility with a target substance and an anticoagulant exist on the surface of the carriers. Processed corpuscles and plasmas are mixed again and sent to an autotransfusion means comprised of an autotransfusion needle 10 and returned to the body of a subject.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a blood processing method and apparatus for removing harmful substances, pathogenic cells, viruses, virus-infected cells and the like in blood.

[0002]

2. Description of the Related Art A conventional blood processing system is disclosed in
-113463, JP-B-5-50302, JP-B-5
No. -50303, Japanese Patent Publication No. 5-50301, etc., after separating a plasma component and a blood cell component using a plasma separation membrane or a centrifugal separator, the plasma component is immunosorbent or low-density lipoprotein adsorbent. It was a system that directly contacted with such as to adsorb and remove the target substance. In these systems,
Since the target substance adsorbent comes into direct contact with the plasma component, it was used while administering an anticoagulant to prevent blood from coagulating.
In addition, there is a problem that the dose of the anticoagulant is increased. Systems for killing and adsorbing and removing viruses and virus-infected cells have also been studied, but all have had effects on blood protein components and blood coagulation problems.

[0003] Many viruses have an isoelectric point of 3 to 6, and have a negative charge in a neutral region. Therefore, viruses can be adsorbed from a buffer or the like by electrostatic interaction. Such a virus removing material is disclosed in
There is a material having a cationic compound on its surface as described in JP 123630. Also, JP-A-2-368.
No. 78 discloses HIV consisting of a solid substance having a weakly acidic or weakly basic surface and a remover for removing the related substance. This remover removes -COOH and -SO
_{It has 3} H or the like, and has a surface pH of 2.5 to 6.9 or 7.
It is 4 to 10.5 have been characterized, when the -COOH or -SO ₃ H or the like on the surface forms a salt,
It is stated that HIV cannot be removed. Furthermore, in the method described in JP-A-2-36878, it has been confirmed that when blood is brought into contact with a removing agent, the pH of blood changes, and denaturation of component proteins occurs, which is not always a favorable condition for blood. Was not. As described above, a problem common to these techniques is that there is a risk of blood coagulation due to denaturation of blood proteins when the material comes into contact with blood.

As a membrane relatively insensitive to blood contact compared to the above-mentioned system, Japanese Patent Laid-Open No. 2-167232 discloses a regenerated cellulose membrane for removing viruses from body fluids and protein solutions by pore size. . This membrane
The pore size is smaller than the HIV particles and does not transmit HIV. However, there is a problem that since the pore diameter is small, the transmission speed is small and clogging is likely to occur.

[0005]

DISCLOSURE OF THE INVENTION The present invention removes or inactivates target substances such as harmful substances, pathogenic cells, viruses, and virus-infected cells from blood, and also improves the problem of blood coagulation and blood pH change. It is an object of the present invention to provide a blood processing system.

[0006]

That is, the present invention provides a method for extracting blood from the body, removing harmful substances, pathogenic cells,
A method of treating blood with a carrier having on its surface a compound that adsorbs, removes or inactivates at least one target substance selected from viruses and virus-infected cells and an anticoagulant, and then returns the blood to the body I will provide a. Further, the present invention has a blood collecting means, a blood processing means connected to the blood collecting means, and a blood returning means connected to the blood processing means, wherein the blood processing means has harmful substances, pathogenic cells, viruses, virus infections in blood. Provided is a blood processing apparatus having a carrier that holds a compound that adsorbs, removes, or inactivates at least one target substance selected from cells, and an anticoagulant on its surface. The compound that adsorbs, removes, or inactivates the target substance is preferably a polyamine compound such as polyethyleneimine, polyallylamine, polyvinylamine, and polypropyleneamine.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION The method of the present invention comprises the steps of removing blood from the body, treating the blood with a carrier having a compound that adsorbs, removes, or inactivates a target substance, and an anticoagulant on the surface thereof, It is a method of returning blood to the body.

The blood treatment carrier used in the method of the present invention can be exemplified by a porous flat membrane, a hollow fiber membrane, a nonwoven fabric, a woven fabric, a porous bead, etc. In the case of a nonwoven fabric, the filament may be a monofilament or a multifilament. It may be a quality filament or an irregular filament. In addition, the material of the carrier is not particularly limited, and natural polymers such as cellulose and derivatives thereof, polyolefins, polyvinylidene fluoride, polyamide, polyimide, polyurethane, polyester, polysulfone, polyacrylonitrile, and other high polymer materials can be exemplified. It is preferable that the surface is modified to be hydrophilic by surface treatment.

When the carrier is a porous membrane, the average pore size is preferably 0.1 to 10 μm. If the pore size is less than 0.1 μm, a sufficient amount of filtration cannot be obtained, and clogging is likely to occur. If the pore size exceeds 10 μm, appropriate removal cannot be performed. When the carrier is in the form of a nonwoven fabric, a nonwoven fabric having a fiber diameter of 100 μm or less on average is preferable. If the fiber diameter is large, it becomes difficult to secure the surface area of the substrate, and it becomes difficult to immobilize a polyamine compound or the like on the surface of the substrate. When the carrier is a porous bead, the average particle size of the porous bead is preferably 25 μm to 300 μm. If the average particle diameter is smaller than 25 μm, the filtration pressure will be high, and if it is larger than 300 μm, the surface area per volume will be small, and it will not be possible to fix sufficient amino groups.

The target substance in the method of the present invention is a harmful substance, a pathogenic cell, a virus, a virus-infected cell or the like in blood. For example, AIDS and blood associated with the cause and progression of AIDS and AIDS-related syndrome are exemplified. HI inside
V, HIV-infected cells, HIV-related substances and complexes with these biological components, and I produced by AIDS progression
They are cytokines such as L-1, IL-6 and TNF, and pathogenic substances such as endotoxin.

As the compound that adsorbs, removes or inactivates the target substance, a polyamine compound is preferable. The polyamine compound is not particularly limited as long as it is a compound having a plurality of amino groups in a molecule, and examples thereof include polyethyleneimine exemplified above, polyallylamine, polyvinylamine, and polypropyleneamine. Among them, polyethyleneimine having particularly high charge density is preferable.

Various known methods can be applied as a method of having the polyamine compound on the surface of the carrier. To have here means to exist on the carrier surface so as not to elute in water or blood, for example, a graft polymerization method,
After introducing a functional group such as an epoxy group, amino group, formyl group, carboxyl group, hydroxyl group, acid halide group, cyanogen halide group, etc. onto the carrier surface by a coating method or the like, the polyamine compound is directly or a coupling agent or spacer. And the like.

The anticoagulant is not particularly limited as long as it is a compound having an anticoagulant activity. Heparin, Fusan, FOY, argatroban, citric acid and the like are preferred examples, and heparin is preferred. Anticoagulants
It releases anticoagulant activity while being sustainedly released or remaining on the surface of the material.

After dissolving the anticoagulant in the solvent, the anticoagulant is coated on the surface of the carrier by a coating method such as coating, spin coating, dipping or spraying.

In the present invention, it is preferable that both the polyamine compound and the anticoagulant are present on the surface of the carrier. It is desirable that the amino group of the polyamine compound and the anticoagulant form a salt with each other, but a free amino group that does not form a salt with the anticoagulant must be present. When the amino group and the anticoagulant do not form a salt, that is, when the amino group remains in a free state, the amino group alone is -CO
OH, and the like are formed with a salt -SO ₃ H or HCl. This is because pH change can be suppressed by forming a salt.

The method of the present invention can be suitably performed by the blood processing apparatus of the present invention. FIG. 1 shows the blood processing apparatus of the present invention.
This will be described using the embodiments shown in FIGS. The blood processing apparatus 1 according to the present invention is capable of detecting viruses, virus-infected cells,
It is a device for removing or inactivating harmful substances and the like, having a blood collecting means, a blood processing means, a blood returning means,
Each device is communicated with a tube. The blood to be processed is collected from the subject via the blood collecting means, then processed by the blood processing means, and returned to the subject again by the blood returning means.

The blood collecting means of the present apparatus has a blood collecting needle 8.
As the blood collection needle 8, a known metal needle or a synthetic resin needle is used. A tube is connected to the needle. The blood collecting means of the apparatus of the present invention may have a blood collecting pump 12 as necessary. When the blood collecting pump 12 is provided, the blood collecting needle 8 and the blood collecting pump 12 are communicated via a tube.

The blood processing means is connected to the blood collecting means, and the blood collected from the subject by the blood collecting means is sent to the blood processing means for processing. The blood processing means includes a plasma separation device 4, a plasma processing device 5, and a blood cell processing device 6.
As shown in FIG. 1 (b), the collected blood is first separated into a plasma component and a blood cell component by a plasma separation device 4, and distributed so that each component is processed by a plasma processing device 5 and a blood cell processing device 6. As shown in FIG. 1 (a), the collected blood is first treated with a blood cell component by a blood cell processing device 6 and then separated into a plasma component and a blood cell component by a plasma separation device 4. Preferably, the plasma component is processed by the plasma processing apparatus 5. This is because the blood cell processing device is likely to be clogged after the plasma is separated. If necessary, a plasma pump 13 may be provided in the blood separation device and the plasma processing device.

As the plasma separating device 4, a hollow fiber type plasma separating device or a centrifugal type plasma separating device is exemplified. In the hollow fiber type plasma separation apparatus, a whole blood component is caused to flow outside the hollow fiber, a plasma component is separated into the hollow fiber, and a blood cell component is separated and taken out from the hollow fiber. The centrifugal plasma separator centrifuges whole blood components to sediment blood cell components, collects the supernatant, and separates the supernatant as a plasma component.

The blood cell processing apparatus 6 is a housing in which a carrier such as a porous flat membrane, a hollow fiber membrane, a non-woven fabric, a woven fabric, or a porous bead is filled in a housing. There are compounds with affinity and anticoagulants.
When the carrier used for the blood cell processing device 6 is a porous body, the average pore size is preferably 1 to 10 μm. This is for processing without clogging. The treatment target is not particularly limited as long as it is a blood cell component, and includes red blood cells, platelets, and white blood cells (neutrophils, basophils, lymphocytes, and monocytes). Among them, white blood cells are preferable for AIDS treatment. .

The plasma processing apparatus 5 is an apparatus in which a housing having an inlet and an outlet for blood is filled with a carrier such as a porous flat membrane, a hollow fiber membrane, a nonwoven fabric, a woven fabric, and a porous bead. When the carrier used for the plasma processing apparatus 5 is a porous body,
The average pore size is preferably from 0.1 to 0.8 μm. This is for ensuring the surface area and processing without clogging. The carrier has on its surface a compound having an affinity for a target substance and an anticoagulant.

The detailed description of the compound for adsorbing, removing or inactivating the target substance, the anticoagulant, and the carrier holding these on the surface used in the apparatus of the present invention is as described in the method of the present invention. .

The housing of each device is made of a synthetic resin such as polypropylene, polycarbonate, polyethylene, polystyrene, polymethacrylic acid, or a metal such as glass or stainless steel.

In the present apparatus, the tubes communicating between the members are:
It is made of a transparent synthetic resin having transparency, such as a vinyl chloride resin and a silicone rubber.

The blood cell component and the plasma component processed by the blood processing means are mixed again, sent to the blood return means connected to the blood processing means, and returned to the body of the subject. The remixing of the blood cell component and the plasma component is performed by flushing the processed blood cell component discharged from the plasma separation device 4 or the blood cell processing device 6 with the processed plasma component. Further, mechanical stirring means may be used. Examples of the mechanical stirring means include stirring with a stirring blade and a magnetic stirrer. However, when using a mechanical stirring means, gently agitate so that blood cell components are not damaged. Blood return means is blood return needle 1
Consists of zero. As the blood return needle 10, a known metal needle or a synthetic resin needle is used.

The blood processing apparatus of the present invention may have the form shown in FIG. In this embodiment, the present apparatus has a temporary blood reservoir 14 between the blood collection means and the plasma separation device 4. By providing the temporary blood reservoir 14, the pressure loss can be adjusted and the flow rate can be kept constant. Temporary blood storage container 14
Is a sealed container made of a soft synthetic resin (for example, soft vinyl chloride), a hard synthetic resin (for example, polycarbonate, hard vinyl chloride), or an open container. ACD liquid as an anticoagulant in the temporary blood reservoir 14;
A CPD solution, sodium citrate, heparin, or the like may be injected in advance.

In the apparatus according to the present invention shown in FIG. 2, the blood collecting means also functions as the blood returning means. Thus, a single needle is sufficient without mounting two needles, a blood collection needle and a blood return needle, on the patient, so that the burden on the patient can be reduced. Further, a valve may be appropriately provided between each means and between each device constituting the blood processing means. By providing the valve, for example, it is possible to adjust the mixing ratio of the blood cell component and the plasma component that have been processed, or to take out only one of the components from the apparatus.

Further, the present apparatus may have, if necessary, means for removing air that has entered the apparatus.

According to the present invention, it is possible to remove and inactivate a target substance or the like in blood by suppressing pH change without coagulating blood. In addition, according to the present invention, blood from which a target substance (target substance, for example, HIV) has been removed can be returned to the body, which is effective in treating diseases corresponding to the target substance. For example, if a subject is suffering from AIDS, human immunodeficiency syndrome virus (HIV) and its related substances are obtained from blood and blood products for the purpose of preventing and treating acquired immunodeficiency syndrome (AIDS). , IL-1, IL- produced by HIV infection
6. It can remove cytokines such as TNF, endotoxin that causes sepsis, and the like. Journal of Clinical Apheresis, 3, 133-139 (1
986) describes that lymphoplasmapheresis, which is a combination of plasmapheresis and lymphocytopheresis, is effective for AIDS treatment. Blood cell processing and plasma processing are performed as a method or apparatus for performing the treatment. The combined method or device is useful for AIDS treatment.

[0030]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described by taking the following blood processing system for AIDS as an example, but the present invention is not limited to this.

A blood processing system was created with the configuration shown in FIG. 1) Blood collection means A tube made of vinyl chloride resin having an inner diameter of 3 mm was connected to a blood collection needle.

2) Blood Cell Processing Apparatus A leukocyte removal filter or the like was used as a blood cell processing apparatus. 3) Plasma separator A commercially available hollow fiber type plasma separator was used as the plasma separator.

4) Plasma treatment apparatus Production of porous membrane An argon plasma (100 W, 0.1 Ton) was applied to a polypropylene porous membrane having an average pore diameter of 0.45 µm and a film thickness of 80 µm.
rr, 15 seconds), and then contacted with a 2-methoxyethyl acrylate gas (1.0 Torr) for 3 minutes with a glycidyl methacrylate gas (0.7 Torr) to perform surface graft polymerization, thereby obtaining hydrophilicity having an epoxy group on the surface. A porous membrane was obtained. Subsequently, the film was immersed in an aqueous solution containing 0.3% by weight of polyethyleneimine (molecular weight: 70,000) and 0.01% by weight of pyridine for 10 seconds, and further immobilized at 90 ° C. for 10 minutes to immobilize the polyethyleneimine on the membrane surface. . The average pore diameter is a value obtained by using a perm porometer (manufactured by Porous Materials Co., Ltd.) based on ASTM-F316.

The membrane prepared in the evaluation of the porous membrane was immersed in a 5 U / ml heparin saline solution for 30 minutes, and then set in a NUCLIpore Sinlock filter holder (φ25 mm).
3 ml of human plasma collected from V-positive patients was filtered.
The presence or absence of HIV infectivity before and after filtration was measured and compared. As a result, the infectivity had disappeared after filtration.

Here, the infectivity was evaluated as follows. The plasma before and after the filtration was serially diluted in two-fold dilutions using RPMI1640 containing 10% FCS, and human lymphocytes were added thereto, cultured, and the presence or absence of infection was determined by HIV antigen (p24).
Antigen).

Production of Plasma Processing Apparatus The obtained membrane was pleated with a width of 24 mm, and then filled in a rectangular polycarbonate housing having a width of 25 mm, a length of 120 mm and a height of 60 mm. The effective film area of this module is about 0.7 m ² .

5) Blood return means A blood return needle was connected to the tip of a tube made of vinyl chloride resin having an inner diameter of 3 mm, and blood was returned from the blood return needle into the body while mixing the blood cell component and the plasma component.

[0038]

According to the blood processing method and apparatus of the present invention, a blood processing means is provided between a blood collecting means and a blood returning means, and the blood processing means comprises harmful substances, pathogenic cells, viruses,
A plasma processing apparatus and a blood cell processing apparatus including a carrier that adsorbs, removes, or inactivates at least one target substance selected from virus-infected cells, and a carrier having an anticoagulant on its surface, In the case that the blood of the patient containing the target substance is processed, the plasma processing apparatus and the blood cell component processed by the blood cell processing apparatus have means for mixing and returning the blood. It is possible to remove or inactivate a target substance or its related substance from plasma containing the target substance or its related substance, and remove a blood cell component having an antigen to be a target substance by a blood cell processing apparatus. . Further, if the compound that adsorbs, removes or inactivates the target substance is a polyamine compound, it forms a salt with an anticoagulant, so that blood coagulation and blood denaturation due to pH change can be suppressed. Take AIDS treatment as an example.
Reduction of viral load, prevention of virus reinfection, and prevention of virus replication are important for DS treatment. In other words, from the body fluid of a patient with HIV blood, HIV or HIV
Removing related substances or HIV infected cells
It is effective in improving QOL of patients and suppressing progression of disease.

[Brief description of the drawings]

FIGS. 1A and 1B are views showing a two-needle type embodiment of the blood processing apparatus of the present invention.

FIG. 2 is a diagram showing a single-needle mode of the blood processing apparatus of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Blood processing apparatus 4 Plasma separation apparatus 5 Plasma processing apparatus 6 Blood cell processing apparatus 7 Blood flow 8 Blood collection needle 10 Blood return needle 12 Blood collection pump 13 Plasma pump 14 Temporary blood storage container 15 Valve

Claims (4)

[Claims] 1. A compound which removes blood from the body, adsorbs, removes or inactivates at least one target substance selected from harmful substances, pathogenic cells, viruses and virus-infected cells in blood, and an anticoagulant And treating the blood with a carrier having on the surface thereof and then returning the blood to the body. 2. A blood collecting means, a blood processing means connected to the blood collecting means, and a blood returning means connected to the blood processing means, wherein the blood processing means comprises harmful substances, pathogenic cells, viruses and virus-infected cells in blood. A compound that adsorbs, removes, or inactivates at least one target substance selected from:
A blood processing apparatus having a carrier having on its surface an anticoagulant. 3. The blood processing apparatus according to claim 2, wherein the compound that adsorbs, removes, or inactivates the target substance is a polyamine compound. 4. The blood processing apparatus according to claim 3, wherein the polyamine compound is any one of polyethyleneimine, polyallylamine, polyvinylamine, and polypropyleneamine.