JPH0513696B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a blood purification adsorbent for removing malignant substances and malignant cells from blood and plasma, and particularly to an adsorbent suitable for whole blood blood purification using a direct perfusion method. More specifically, diseases and phenomena related to the body's immune system such as cancer, immunoproliferative syndrome, rheumatoid arthritis, systemic lupus erythematosus, allergies, rejection reactions during organ transplants, kidney diseases such as nephritis, hepatitis, etc. The present invention relates to an adsorbent that adsorbs and removes malignant substances and malignant cells from body fluids such as blood and plasma that are thought to be closely related to the cause or progression of the disease in liver diseases.

Conventionally, activated carbon or activated carbon coated with a hydrophilic polymer has been used as an adsorbent for body fluid purification treatment, mainly as an artificial liver for liver diseases. However, in many diseases, various malignant substances and malignant cells that are closely related to the cause or progression of the disease have become known, and there is a growing need to selectively remove these malignant substances and malignant cells from body fluids. However, adsorbents based on activated carbon are currently unable to meet this demand because the substances they can adsorb are limited.

Furthermore, in body fluid purification treatments using adsorbents, blood coagulation induced by contact between adsorbents such as activated carbon, which are foreign substances to living organisms, and body fluids such as blood has been a major problem. In order to prevent this blood coagulation, attempts have been made to coat activated carbon with hydrophilic polymers, but sufficient antithrombotic properties have not been obtained, and heparin, an anticoagulant, is supplied into the blood. The current situation is that it prevents blood coagulation. In this case, since a large amount of heparin is used, there are problems such as side effects such as bleeding in some cases.

As a result of intensive research in response to the demand for selective adsorption and removal of malignant substances, the present inventors have found that the carrier retains a special substance that selectively interacts chemically with the adsorbed substance through chemical bonds. Various adsorbents have been discovered (Japanese Patent Publication No. 63-53971, Japanese Patent Publication No. 57
−192560, Special Publication No. 1-34067, Japanese Unexamined Patent Publication No. 1983-
(Refer to each publication No. 134164). The present invention was achieved as a result of further research into the above-mentioned blood coagulation problems.

That is, in view of the problem of blood coagulation due to contact between an adsorbent and blood based on the conventional technology as described above, the present inventors have developed a method for selectively adsorbing malignant substances and malignant cells in body fluids and by contacting them. As a result of intensive research on adsorbents that do not coagulate blood, we have discovered that an organic compound (hereinafter referred to as an organic ligand) that has a polyanion graft chain and a functional site that can bind to the adsorbed substance on the surface of an insoluble carrier. The present inventors have discovered that an adsorbent formed by bonding the above can solve the above-mentioned problems, and have completed the present invention.

Considering the two functions of platelet adhesion inhibition and affinity-type adsorption ability, which have not been easily solved in the past, we have developed an organic ligand that has affinity for malignant substances and malignant cells in the blood, while binding it to a carrier. It is necessary to provide a means for inhibiting platelet adhesion on the same carrier, and the present inventors have discovered the binding of polyanion graft chains as an excellent means for inhibiting platelet adhesion. This is expected to make platelets more difficult to approach due to the electrostatic repulsion of platelets due to their negative charge and the whisker effect (excluded volume effect) on the carrier due to the hydrophilic graft chain, and this effect is actually observed. I was able to confirm that.

As the insoluble carrier in the present invention, when the malignant substance is a soluble body fluid component, it is preferable to use a porous carrier, particularly a porous polymer. The exclusion limit molecular weight (protein) of the porous polymer used in the present invention corresponds to the size of the malignant substance to be adsorbed, and should be at least 20,000 or more to effectively purify blood for the purpose of the present invention. It is necessary to achieve this. When the adsorbed substance is an immune complex, especially an IgM immune complex, the number reaches 10 million.
10 million is preferred. The most common exclusion limit molecular weight for purposes of this invention is 100,000 to 5,000,000. The specific surface area of the insoluble carrier in the present invention is preferably 5 m ² /g or more, more preferably 55 m ² /g or more. There are various methods for measuring the specific surface area, but in the present invention, it was determined by the most common BET method using nitrogen gas. In addition, the sample used for specific surface area measurement must be sufficiently dry, but if the carrier is difficult to dry, after equilibrating the wet carrier with acetone,
The sample was dried under reduced pressure at 60°C or lower to prepare a measurement sample.

In the present invention, the insoluble carrier is particulate, fibrous,
Any known shape such as hollow fiber, membrane, or sheet shape can be used, but in terms of the amount of organic ligand retained and ease of handling as an adsorbent, particulate,
Fibrous materials are preferred, and particularly granular materials give favorable results.

The particle size of a spherical or particulate carrier is determined from its specific surface area (adsorption capacity as an adsorbent) and body fluid circulation.
A thickness of 50 to 1500 μm is preferable. A more preferable range is 200 to 1000 μm.

Further, when a fibrous carrier is used, the fiber diameter thereof is preferably in the range of 0.02 denier to 10 denier, more preferably 0.1 denier to 5 denier. If the fiber diameter is too large, the adsorption amount and rate of globulin compounds will be reduced, and if it is too small, activation of the coagulation system, blood cell adhesion, and clogging may occur.

In order to immobilize the ligand, the above-mentioned insoluble carrier only needs to be able to be activated and to be physically stable throughout the entire process including the carrier activation reaction, immobilization reaction, adsorption operation, etc. It can be selected from either hydrophilic compounds or hydrophobic compounds. Specifically, for inorganic-based products,
Activated carbon, glass, etc. and their derivatives are available, and natural polymer-derived carriers include simple polysaccharides and their derivatives such as cellulose, Cephalose, dextran, starch, alginic acid, and chitin, and complex polysaccharides such as agar, pectin, konjac, and gum arabic. There are sugars and their derivatives, proteins such as wool and silk proteins, and their derivatives.
After undergoing insolubilization treatment such as crosslinking reaction, it is used as a carrier.

Regarding synthetic polymers, vinyl polymers include styrene, vinyl acetate, methacrylate ester, acrylate ester, vinyl halide,
There are polymers and copolymers of vinylidene halides, acrylonitrile, acrylamide, ethyl vinyl ketone, vinylpyrrolidone, 2-vinylpyridine, ethylene, propylene, butadiene, isobrene, etc. and their derivatives, and ring-opening polymers of cyclic compounds include , dimethylcyclopropane, spiro-di-o-xylylene, norbornene, cyclobutene, trioxane, lactide, cyclopolysiloxane, phosphonitrile chloride, N-carboxy-
Polymers and copolymers of α-amino acid anhydrides, etc. and their derivatives, polyformaldehyde, polyethylene oxide, polypropylene glycol, poly-3,3-bis(chloromethyl)oxacyclobutane, polytetrahydrofuran, polycaprolactam, etc., and their derivatives. .

Examples of the polycondensate include polyester, polyamide, polyanhydride, polycarbonate, polyurea, polysulfonamide, polyimide, polybenzimidazole, and derivatives thereof.

For resins and other materials, acrylic resin,
Examples include methacrylic resin, fluororesin, epoxy resin, urea resin, amino resin, styrene resin, melamine resin, polyurethane, silicone resin, alkyd resin, and derivatives thereof.

The above-mentioned polymer carriers can be made into insolubilized carriers by using appropriate comonomers and crosslinking agents as necessary. , epoxy compounds, dienes, glutaraldehyde, etc. can be selected according to the functional group of the object to be crosslinked (Taiseisha, "Crosslinking Agent Handbook", p.3-77,
1981).

Among these insoluble carriers, since adsorbents are created that utilize the affinity of the ligand immobilized on the carrier, hydrophilic carriers are generally used since the carrier itself must not non-specifically adsorb biological substances. Preferably, it is a sexual carrier.

Further, since increasing the immobilization capacity of the ligand is advantageous in increasing the adsorption capacity, it is preferable to use an organic polymer carrier that can easily obtain a high activation rate carrier. Furthermore, it is preferable to use an organic synthetic polymer carrier that can control mechanical and/or hydrodynamic properties more appropriately. That is, a hydrophilic organic synthetic polymer provides a more preferable carrier. Among such carriers, polymers or copolymers containing vinyl alcohol units are more preferred carriers because of their ease of handling and wide applicability.

In addition, in the case of polymers or copolymers containing vinyl alcohol units, compounds having a triazine ring such as carboxylic acid vinyl ester and triallyl cyanurate, triallyl isocyanurate, ethylene glycol dimethacrylate, diethylene glycol dimethacrylate, etc. Di(meth)acrylates, butanediol divinyl ether,
A crosslinkable monomer selected from polyvinyl ethers such as diethylene glycol divinyl ether and tetravinylglyoxal, and polyallyl ethers such as diarylidene pentaerythrite and tetraallyloxyethane is copolymerized and the copolymer is esterified. Insoluble carriers obtained by exchange or saponification are preferred from the viewpoints of strength, micropore structure, and chemical stability.

The polyanion graft chain retained on the carrier of the present invention exhibits a negative charge in a neutral electrolyte such as blood or body fluid while bound to the carrier. In the present invention, the negative charge density of the polyanion graft chain is at least
A concentration of 0.02 μeq/m ² can be expected to have the effect of reducing interaction with platelets. Furthermore, the preferred range of negative charge density is
The range is from 0.2 μeq/m ² to 2500 μeq/m ² , and a more preferred range is from 0.5 μeq/m ² to 500 μeq/m ² .

In the present invention, in the case of a multiphase structure in which the negative electrode is localized on the carrier surface, it is sufficient that the negative charge density in the surface layer is within the above range.

The molecular weight of the polyanion graft chains retained on the carrier of the present invention is preferably 600 daltons or more on average in number, in view of the excluded volume effect of the graft chains. In addition, a more stable platelet adhesion suppressing effect can be expected, and the molecular weight range that does not impede adsorption ability is
It ranges from 1,500 to 1,000,000 Daltons.

To illustrate the above polyanion graft chains, as negative electrode functional groups, sulfonic acid groups, carboxylic acid groups, phosphoric acid ester groups, etc. are preferably used, and specific examples include polygalacuronic acid,
Phosphorylated mannan, chondroitin, chondroitin sulfate A, chondroitin C, hyaluronic acid,
Those based on acidic polysaccharides such as alginic acid, pectic acid, and fucanic acid, and natural polymers include carboxymethyl cellulose, cellulose sulfate, carboxymethyl starch, cellulose phosphate, and their derivatives, and those obtained by polymerization methods. Examples include polyacrylic acid, polymethacrylic acid, polyvinyl sulfuric acid, polystyrene sulfonic acid, maleic acid copolymer, polyphosphoric acid, polyvinyl phosphoric acid, polystyrene phosphoric acid, polyglutamic acid, polyaspartic acid, polyphosphate ester, etc., and their derivatives. However, the present invention is not limited to the above substances as long as they are polyanionic polymers.

The organic ligand to be supported on the carrier of the present invention can be freely selected depending on the purpose, and some of them are exemplified below.

For the treatment of systemic lupus erythematosus, adenine,
Homopolymers or copolymers of mono-, di-, and trinucleotides such as guanine, cytosine, uracil, and thymine, naturally occurring nucleic acids such as DNA and RNA, as well as nucleobases, sugars, oligosaccharides, and the like can be used. Also, the DNA present in the blood,
Anti-single-stranded DNA antibody, anti-double-stranded DNA antibody, anti-RNA antibody, anti-ENA for adsorption and removal of RNA and ENA.
Anti-nucleic acid antibodies such as antibodies, basic compounds such as methylated albumin actinomycin D, etc. can be used. Furthermore, for the adsorption and removal of immune complexes in the blood, complement components such as Clq, specific proteins such as protein A, anti-snake chain constant region 2 phase antibodies,
Antibodies against immune complexes such as rheumatoid factor can be used.

For the treatment of rheumatoid arthritis and malignant rheumatoid arthritis, chemical denaturation (coagulation) methods such as urea, guanidine hydrochloride, mercaptoethanol, surfactants, and organic solvents, physical denaturation (coagulation) methods such as heat, ultrasound, and gas bubbling are used. ) Modified γ modified by method
-Globulin, modified immunoglobulin, aggregated γ-
Antigen-like substances against rheumatoid factors, such as globulins, aggregated immunoglobulins, Fc regions of immunoglobulins, second phase of immunoglobulin heavy-chain invariant regions, and modified forms thereof by the above-mentioned denaturation methods, anti-rheumatoid factor antibodies, and tryptophan;
Hydrophobic organic compounds such as phenylalunine can be used. In addition, for the removal of rheumatoid immune complexes, complement components such as Clq, specific proteins such as protein A, anti-heavichain constant region 2 phase antibodies,
Antibodies against immune complexes such as rheumatoid factor can be used.

For the treatment of Hashimoto's disease, thyrogloprine, a microsomal fractionated component of the thyroid gland, can be used.
For the treatment of myasthenia gravis, neuromuscular acetylcholine receptor fraction components can be used.

Glomerular basement membrane components are used to treat glomerulonephritis, platelet membrane components and platelet granule fraction components are used to treat idiopathic thrombocytopenic purpura, and transcortisone and anticortisone antibodies are used to treat Cushing syndrome. be able to.

For the prevention and treatment of hepatitis, hepatitis A virus,
Antibodies against viral surface antigens such as hepatitis B virus can be used.

For hypertension treatment, anti-angiotensin antibodies,
Heparin and anti-riboprotein antibodies can be used to treat hyperlipidemia.

For the treatment of immune diseases based on lymphocyte abnormalities, anti-lymphocyte antibodies such as anti-B cell antibodies, anti-sublesser T antibodies, and anti-helper T antibodies, anti-monocyte antibodies, anti-natural killer cell antibodies, and anti-granulocyte antibodies are used. Can be used.

Anti-erythrocyte antibodies and anti-platelet antibodies can be used for membrane diseases of red blood cells and platelets.

For cancer treatment such as breast cancer, protein A, anti-immunoglobulin antibodies, and antibodies against immunosuppressive factors can be used.

Ligands that can be used in the present invention are not limited to the above examples, but include lectins such as congnitinin, concanavalin A, and phytohemaagglutinin, nucleic acids, amino acids, lipids, glycolipids, protamine, heparin, antigens, antibodies, enzymes,
Known substances capable of binding to adsorbed substances such as substrates, coenzymes, and glycoproteins can be used.

Furthermore, the carrier of the present invention can also be used with two or more types of ligands retained therein. Furthermore, two or more types of carriers holding ligands can be used in combination.

In the present invention, any known method such as covalent bonding, ionic bonding, physical adsorption, embedding, or precipitation insolubilization on the surface of the carrier can be used to immobilize the organic ligand on the surface of the insoluble carrier. From the viewpoint of elution properties, it is preferable to use it after it is fixed and insolubilized by covalent bonding.
For organic ligands, conventional immobilized enzymes, known methods for activating insoluble carriers used in affinity chromatography, and methods for binding to ligands can be used. Furthermore, if necessary, a molecule (spacer) of arbitrary length can be introduced between the insoluble carrier and the ligand.

Examples of functional groups that can form covalent bonds include hydroxyl group, amino group, carboxyl group, epoxy group, chloroformyl group, azide group, formyl group, bromoacetyl group, isocyanate group, cyano group, acid chloride group, and acid chloride group. Examples include anhydride groups and diazocoum groups, but if these functional groups have low reactivity and it is difficult to directly form a covalent bond with a ligand, highly reactive functional groups can be activated using known activation methods. After conversion, a covalent bond may be formed with the ligand. For example, the hydroxyl group of the carrier can be activated by a cyanogen halide method, an epichlorohydrin method, a bisepoxide method, a halogenated triazine method, a bromoacetyl bromide method, an ethyl chloroformate method, a carbonyldiimidazole method, or the like. These functional groups are amino groups of organic ligands,
It can react with nucleophilic reactive groups such as hydroxyl groups, carboxyl groups, and thiol groups to form covalent bonds.

On the other hand, methods for bonding polyanion graft chains to the surface of an insoluble carrier include methods similar to the covalent bonding and ionic bonding in the organic ligands described above, as well as oxidative graft polymerization using oxygen, ozone, cerium salt, etc. Examples include graft polymerization using an organic peroxide such as bisisobutyronitrile or benzoyl peroxide, and graft polymerization of an anionic monomer onto a carrier such as radiation graft polymerization. Examples include a method of coupling.

The method for producing an adsorbent according to the present invention, which is comprised of the above-mentioned elements, does not specify the order in which the constituent elements are combined. Specifically, a carrier or adsorbent obtained by introducing a polyanion graft chain into a carrier or adsorbent that does not have a polyanion graft chain also represents one embodiment of the present invention.
Furthermore, the bonding order of the polyanion graft chain and the organic ligand can also be selected depending on the difficulty in manufacturing. Furthermore, in the method of introducing an organic ligand, the purpose can be achieved by a method in which the organic ligand is bonded to a monomer and polymerized, or a simultaneous reaction is performed during the formation of an insoluble carrier by post-crosslinking of a polymeric substance.

That is, the present invention is not dependent on the manufacturing method, as long as the polyanion graft chain and the organic ligand are present in the adsorbent.

The adsorbent of the present invention can be used by being filled and held in a container equipped with an inlet and outlet for body fluids.

The drawing shows an example of an adsorption device using the adsorption body of the present invention, in which a cap 6 having a body fluid inlet 5 is screwed into an opening at one end of a cylinder 2 through a packing 4 with a filter 3 stretched inside. mated,
A cap 8 having a body fluid outlet 7 is screwed into the opening at the other end of the cylinder 2 through a packing 4' with a filter 3' stretched inside to form a container, and the cap 8 is sucked into the gap between the filters 3 and 3'. The adsorbent layer 9 is formed by filling and holding the adsorbent.

The adsorbent layer 9 may be filled with the adsorbent of the present invention alone, or may be mixed or laminated with other adsorbents. As other adsorbents, for example, activated carbon having a wide range of adsorption capacities can be used.
As a result, a wider range of clinical effects can be expected due to the synergistic effect of the adsorbent. The appropriate volume of the adsorbent layer 9 is about 50 to 400 ml when used for extracorporeal circulation.

When the device of the present invention is used for extracorporeal circulation, there are roughly two methods as follows. One method is to directly pass blood taken from the body through the device for purification, and the other is to use a centrifuge or membrane plasma separator to purify blood taken from the body. This is a method in which specific components are separated and then malignant substances and malignant cells contained in those components are removed. At this time, the method of passing body fluids may be continuous or intermittent, depending on clinical needs or equipment conditions.

As described above, the adsorbent of the present invention can be applied to general uses for purifying and regenerating body fluids such as autologous blood and autologous plasma, and is applicable to cancer, immunoproliferative syndrome, rheumatoid arthritis, etc. Collagen diseases such as systemic lupus erythematosus, autoimmune diseases such as myasthenia gravis, allergic diseases such as bronchial asthma, diseases and phenomena related to biological immune function such as organ transplant rejection, psoriasis, and whitening. It maintains high performance without sacrificing the adsorption capacity, which is the basic property of adsorbents, for extracorporeal circulation treatment of skin diseases such as acne, kidney diseases such as nephritis, liver diseases such as hepatitis, and blood diseases. It is a blood purification adsorbent that can perform blood purification of whole blood through direct extracorporeal circulation, as it exhibits excellent antithrombotic properties.

Embodiments of the present invention will be explained in more detail with reference to Examples below.

Example 1 A homogeneous liquid mixture consisting of 100 g of vinyl acetate, 41 g of triallyl isocyanurate, 100 g of ethyl acetate, 7 g of polyvinyl acetate (degree of polymerization 500) and 3.6 g of 2,2'-azobisisobutyronitrile, and 1 g of polyvinyl alcohol % by weight, 0.05% by weight of sodium dihydrogen phosphate dihydrate and 0.4% by weight of water containing 1.5% by weight of disodium hydrogen phosphate dodecahydrate were placed in a flask and heated at 65°C for 18 hours and then at 75°C. 5
Suspension polymerization was carried out by heating and stirring for a period of time to obtain a granular copolymer. This polymer was hydrolyzed with caustic soda to obtain a polyvinyl alcohol crosslinked polymer. This polymer has an average particle size of 350 μm and a surface area of 120
m ² /g, and the protein exclusion limit molecular weight was 750,000. Using this polymer, an organic ligand was introduced by the following method. That is, 15 g of dry carrier was mixed with 180 ml of dimethyl sulfoxide and 120 ml of epichlorohydrin.
15 ml of 30% aqueous sodium hydroxide solution was added, and the mixture was stirred at 30°C for 75 hours to obtain an epoxy activated carrier.

The amount of epoxy group bonded in the gel was 35 μmol per 1 ml of gel. Using the epoxy binding gel,
An adsorbent was prepared by binding L-phenylalanine. A solution of phenylalanine dissolved at 0.025 mmol/ml in carbonate buffer of pH 9.8 was prepared, and 40 ml of the solution was added to 20 ml of the epoxy binding gel, followed by reaction at 50° C. for 20 hours. The excess active group is
Blocking was performed with 0.1 mol/glycine. The amount of phenylalanine immobilized was calculated from the absorption at 257.5 nm of phenylalanine remaining in the reaction solution. The amount of phenylalanine bound in the adsorbent is
It was 31 μmol/ml. After thoroughly washing the adsorbent with water, it was washed with physiological saline, dehydrated, and used as an evaluation test subject.

Furthermore, take 10 ml of this adsorbent, freeze-dry it, and then
Graft polymerization of styrene sulfonic acid onto the hydroxyl groups of the adsorbent using ceric ammonium nitrate was carried out by a conventional method. The reaction is styrene sulfonic acid
0.14mol/, cerium nitrate salt 10mmol/ in water
The mixture was dissolved in 250 ml and stirred at 35°C for 3 hours to mix with the adsorbent.

The negative charge density of the obtained polyanion was determined by cation exchange capacity measurement and was found to be 6.0 μeq/m ² . The polyanion graft chain of this adsorbent was prepared in the form of a sodium salt by a conventional method and subjected to evaluation.

This adsorbent was packed into a 3 ml column (L/D=5), and fresh bovine whole blood (heparin-added 500 U/100 ml blood) was passed through the column at 1.0 ml/min for 1 hour each in a single bath method. As a result, no decrease in packing volume, clogging, or decrease in flow rate was observed, and the change in the pressure gauge in front of the column was 10 to 20 mmHg.

The adsorption experiment was carried out by mixing 3 ml of chronic rheumatism patient plasma and 1 ml of the adsorbent and incubating the mixture at 37°C for 2 hours. For analysis of rheumatoid factor, adsorption reaction supernatant was used with RAHA Chist (HA No. 3) [Fujirebio
Co., Ltd.] and RA (KW) Kituto [Kyowa Pharmaceutical Co., Ltd.]
Positive or negative results were determined based on the presence or absence of an agglutination reaction that occurred when mixed with a serially diluted solution of patient plasma, and the antibody titer was determined using the highest dilution factor that gave a positive result. The antibody titer of patient plasma is RA160,
The antibody titer of the adsorbent according to this example was 20.80 against RAHA1280.

On the other hand, a similarly prepared adsorbent without polyanion graft chains resulted in a decrease in platelets. Also,
The antibody that did not bind phenylalanine had an antibody titer of RA80 and RAHA320 in the same patient's plasma, and did not have good rheumatoid factor adsorption ability.

Example 2 As a vinyl compound-based porous polymer, an experiment was conducted using a terpolymer consisting of hydroxyl ethyl methacrylate-ethylene glycol dimethacrylate glycidyl methacrylate with an average particle size and an average pore size of 350 μm. I went. The epoxy density in the polymer is 1 mol%. D-(+)-lactose (4-o-β-
β to D-galactopyranosyl-D-glucose)
β-(p-aminophenyl)-ethylated D- reacted with -(p-aminophenyl)-ethylamine
(+)-Lactose was used, and as the polyanion graft chain, one with an average molecular weight of 50,000 obtained by radical telomerization of acrylic acid monomer and 2-aminoethanethiol by a conventional method was used (Journal of the Chemical Society of Japan, 1977 , (1), p88-92). 200 mg of the former organic ligand and 1.0 g of the latter one-terminated amine/polyacrylic acid were competitively reacted with 20 ml of the polymer to obtain adsorbents fixed with 2.7(d) ml of resin and 3.0 mg/ml of resin, respectively.

At this time, the negative charge density of the polyanion graft chain was 1.5 μeq/m ² and the average molecular weight was 15,000 Daltons. As in Example 1, a column flow test using fresh bovine whole blood was conducted, and no thrombus occurred, and the pressure gauge in front of the column was 5 mmHg or less, indicating stable flow.

The adsorption experiment was performed on systemic lupus erythematosus patient plasma 5
and 1 volume of adsorbent were mixed and incubated for 3 hours. The amount of anti-T cell antibodies in the plasma after adsorption was measured by a fluorescent antibody method using normal T cells as a detector. The measurement operations and conditions are as follows.

1. Separate lymphocytes from peripheral blood of healthy individuals by specific gravity centrifugation, and collect T cells by nylon wool column method [“Lymphocyte Function Search Method” Chugai Medical Co., Ltd.
p15, p51 (1980)].

2. Add 0.1 ml of plasma after the adsorption experiment to ¹ × 10 ⁶ isolated healthy human T cells, and treat at 4°C for 1 hour.

3. After washing, apply FITC-labeled anti-human immunoglobulin rabbit antiserum and count fluorescence-positive cells.

4 Express this as a percentage of fluorescence-positive cells.

The results showed that the patient's plasma before adsorption was 23.6%, and the plasma treated with this adsorbent was 0.3%. In addition, the globulin and albumin levels in each plasma were measured using an A/G test (A/GB test, Wako Pure Chemical Industries, Ltd.).
There were no major differences.

On the other hand, the amount of anti-T cell antibodies in plasma treated with an adsorbent that does not bind the organic ligand D-(+)-lactose but only contains polyanion graft chains is
At 22.8%, almost no systemic lupus erythematosus anti-lymphocyte antibodies were adsorbed. Furthermore, in the case of an adsorbent bonded only to D-(+)-lactose without polyanion graft chains, a decrease in platelets was observed in a column flow test using fresh bovine blood.

[Brief explanation of the drawing]

The drawing is a sectional view showing an example of the form of an adsorption device using the blood purification adsorption body of the present invention. 1... Adsorption device, 2... Cylinder, 3, 3'... Filter, 4, 4'... Packing, 5... Body fluid inlet, 6... Cap, 7... Body fluid outlet, 8
... Cap, 9 ... Adsorbent layer.

Claims (1)

[Claims] 1 The polyanion graft chain and the organic compound having a functional site capable of binding to the adsorbed substance are each directly bonded to the surface of the insoluble carrier, and the polyanion graft chain is bonded to the surface of the insoluble carrier through a chemical bond. A blood purification adsorbent that is characterized by the ability to