JPH0412986B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a blood processing device for removing leukocyte components from blood. More specifically, the present invention relates to a blood processing device that can safely and effectively remove leukocytes, including lymphocytes, from blood by extracting and processing a concentrated leukocyte component from blood.

In recent years, with the remarkable development of so-called artificial organs, therapeutic methods have been put into practical use that perform operations such as dialysis, fractionation, or adsorption on blood taken out of the body through extracorporeal circulation, replacing the functions of the kidneys and liver. In addition, it is increasingly being applied to collagen diseases such as rheumatoid arthritis and systemic lupus erythematosus, autoimmune diseases, allergic diseases, and malignant tumors, in which it is clear that immune reactions are deeply involved. The importance of treatment methods is beginning to be recognized.

However, conventional extracorporeal circulation techniques for diseases involving immune reactions only remove substances produced based on abnormal immune reactions, such as immunoglobulins and their aggregates, immune complexes, or cold aggregates. However, it was not effective in correcting abnormal immune responses.

On the other hand, recent advances in immunology have revealed that the main carriers of these immune reactions are white blood cells, especially lymphocytes, which exist in small amounts in the blood, and treatments have been proposed to remove large amounts of white blood cells. ,
Thoracic tube drainage was performed, but there were problems such as the high risk of infection and the difficulty of repeating the procedure, and in order to improve these problems, there was a strong demand for frequent removal of white blood cells from the blood. Ta.

A leukocyte removal filter is known as a separator for separating white blood cells from blood, but this filter only collects granulocytes and monocytes, which have relatively strong stickiness among the white blood cell fraction. It was not possible to collect relatively weak lymphocytes. In addition, when the collection ability is increased, a large amount of platelets are collected at the same time as lymphocytes, resulting in side effects such as bleeding tendency, which precludes frequent use.

In order to overcome the above-mentioned problems and safely and effectively remove leukocytes including lymphocytes from blood, the inventors of the present invention have conducted various studies to extract and process leukocyte-enriched components from blood. I got the idea that
After many more experiments, the present invention was finally completed and the intended purpose was achieved.

That is, the present invention provides a blood processing apparatus for removing leukocyte components from blood, in which a leukocyte concentration means and a leukocyte concentration means for removing leukocyte components from the concentrated leukocyte concentration component are provided between a blood introduction means and a processed blood extraction means. A blood processing apparatus characterized by having a separation means.

Next, a preferred method for concentrating leukocyte components will be described. Blood consists of blood components [red blood cells, white blood cells (granulocytes, lymphocytes, monocytes, monocytes), platelets] and plasma components.When placed in a gravitational field or centrifugal force field, those with higher specific gravity will sediment, and under normal separation conditions The cells are separated from the lower layer into a red blood cell layer, a granulocyte layer, a lymphocyte/monocyte layer, and a platelet-rich plasma layer. Here, the white blood cell layer (granulocyte layer - lymphocytes,
The monocytic layer) is usually called the buffy coat layer.
Among white blood cells, granulocytes tend to sediment due to their larger specific gravity or cell size compared to lymphocytes and monocytes. A large proportion of it is mixed into the layer. Therefore, lymphocytes and monocytes that are more strongly involved in immune reactions tend to be concentrated in the buffy coat layer. After separating the blood into red blood cell concentrated component, white blood cell concentrated component, and platelet-rich plasma component using normal blood component separation technology that processes blood continuously or intermittently in a gravity field or centrifugal force field,
By extracting concentrated leukocyte components and processing them using leukocyte separation means, we can minimize the decrease in platelets and increase the proportion of lymphocytes and monocytes, which are immune cells, compared to granulocytes, which are involved in the defense against infection. It has a more favorable tendency.

In the present invention, the leukocyte-enriched component refers to a leukocyte component in which the concentration of leukocyte components in the fractionated liquid is relatively increased compared to that in the blood as a result of separating the leukocyte component from blood, and the concentration of leukocyte components is determined by the fractionation means. varies somewhat. When practicing the present invention, this white blood cell concentration is
Preferably approximately 100% relative to the concentration in blood.
Double the amount. In addition, leukocyte components include those containing granulocytes, lymphocytes, and monocytes.

Hereinafter, the blood processing apparatus of the present invention will be explained based on the drawings.

FIG. 1 and FIG. 2 are schematic diagrams showing the main parts of an example of the blood processing apparatus of the present invention. In FIG. 1, blood 7 enters leukocyte concentration means 2 from blood introduction means 1, blood components excluding leukocyte concentration component 9,
That is, the concentrated red blood cell component 8, the plasma component 10, and the concentrated white blood cell component 9 that has passed through the white blood cell separating means 3 are each derived from the treated blood deriving means 5. The blood 7 is sent by a pump 6 placed before the blood introduction means 1 or after the blood extraction means 5, as required. FIG. 2 shows the blood components that are mixed by the mixing means 4 to return the viscosity and other properties to the same level as the original blood, and then extracted from the processed blood deriving means 5. Since the transport distance can be shortened, hemolysis and other undesirable phenomena are less likely to occur. It is also possible to use so-called plasma exchange by discarding the plasma component and introducing a plasma substitute into the mixing means. Here, it is desirable that the mixing means be capable of sufficient mixing by stirring, but it may be sufficient to stir the treated blood components by the force of the flow when they are combined. In addition, in FIG. 1 and FIG. 2, 11 is a treated leukocyte concentration component;
12 indicates treated blood.

The leukocyte separation means of the present invention uses fibrous materials such as synthetic fibers and natural fibers, spherical materials such as synthetic polymer beads and glass beads, and porous materials such as polyurethane foam as separation materials to separate blood inlets and outlets. It is a separation means for collecting or removing the target blood. In order to explain in detail the structure of a leukocyte separation means using a fibrous substance by giving an example, one specific example is shown in FIG.

The separation material 13 is filled in a cylinder 14, and a mesh 16 whose periphery is welded and fixed with a ring 15 is provided at both end openings of the cylinder 14, and silicon packing 17 and 18 provided with the ring 15 sandwiched therebetween. It is designed to be fixed by being tightened. Then, in the center is the leukocyte concentration component introduction/extraction tube 1.
A lid 20 provided with a hole 9 is placed over the mesh 16 from the outside, and is tightened and fixed together with the mesh 16 by tightening rings 21 screwed onto the outer periphery of both ends of the cylinder 14.

As the separating material, it is preferable to use fibers of 10 μm or less and packed with a packing density of 0.1 g/cm ² or more.

The average fiber diameter (Dcm) referred to here is defined by the following formula from the weight (xg), length (ycm) of a single fiber, and material density (pg/cm ² ). .

In this case, the cross section of the fiber is generally circular, but the cross section may be any of the various non-circular cross sections found in natural cotton fibers and artificial fibers.

There may be two or more leukocyte separation means, and if a separation means for separating monocytes and granulocytes and a separation means for separating all white blood cells including lymphocytes are combined, lymphocytes, monocytes, and granulocytes can be separated respectively. Can be separated separately.

In other words, in the case of white blood cells that tend to adhere to fibers, such as monocytes and granulocytes, the fiber diameter is 10~
In the case of whole white blood cells including lymphocytes, 60 μm fibers packed at a packing density of 0.04 to 0.4 g/cm ² are used.
A container filled with fibers with a diameter of 10 μm or less at 0.04 to 0.4 g/cm ² is used. This difference in fiber diameter is due to the difference in the adhesion of white blood cells to monocytes, granulocytes, and lymphocytes, and since lymphocytes have weak adhesion, their capture rate will be low unless the fiber diameter is 10μ or less. becomes very low. In the case of a leukocyte separator for the purpose of separating monocytes and granulocytes, a more desirable configuration of the separation means is one with a fiber diameter of 10 to 10.
In the case of a leukocyte separation means that aims to separate white blood cells including lymphocytes, fibers with a fiber diameter of 5 to 8 μm are packed at a packing density of 0.1 to 0.18 g/ ^cm2 . It is packed with a packing density of ^cm2 .

Regarding separation materials other than fibrous materials, lymphocytes, monocytes,
The adhesion of granulocytes can be controlled.

The leukocyte concentration means of the present invention may be any device, regardless of its structure or principle, as long as it is a device that continuously or intermittently introduces blood into a container and causes the blood to settle by the action of centrifugal force or gravity. However, blood is introduced continuously from one side, flows through one or more thin flat sealed containers, and then the red blood cell concentrate component, leukocyte concentrate component, and plasma component are separated from the other side. Particularly preferably used are devices that allow continuous extraction and continuous centrifuges that do not have sliding parts. For example, "Online Plasmaphaeresis of Complete Blood" (W. Ito, J. E. Saudow, R. L. Bowman; Science (USA), vol. 189, p. 999, 1975).
An apparatus such as that described in can be used. namely, a rotating body containing a blood container and a rotating body holder, which, along a flexible cable, include a flexible blood inlet conduit, at least one flexible blood cell enrichment blood outlet conduit, and a flexible blood cell concentrating blood outlet conduit. a plasma outlet conduit, one end of which is coupled to the rotating body along the rotating axis, an intermediate portion bypassing the rotating body, and the other end coupled to the stationary system along the rotating axis on the opposite side of the coupled end. A continuous centrifugation device having a structure in which the inlet conduit and the outlet conduit are connected to the container so that they can communicate with each other by rotating the rotating body and the rotating body holder at a speed ratio of 2:1 by a driving device. A separator is used.

However, in a blood processing device, in order to prevent undesirable side effects such as infection, it is preferable that the parts that come into contact with blood etc. be disposable or replaceable. It became difficult to replace blood containers, etc. The inventors of the present invention have been able to improve the above-mentioned drawbacks by making the blood container and the fluid passage integrated therewith a single assembly that can be easily attached to or detached from the main body of the apparatus. That is, a rotating body containing a blood container is rotatably mounted around a predetermined rotational axis, and both ends of a flexible cable that can be easily bent and cannot be twisted are connected to the rotating body. It is arranged so as to be located approximately on the axis of rotation, and one end is connected to the stationary system with one end facing away from the rotating body, the rotating body is bypassed in the middle, and the other end is connected to the opposite side from the stationary system. a flexible blood inlet conduit and at least one flexible hemoconcentrate blood outlet conduit and a flexible plasma outlet conduit coupled to the rotating body from the cable along the cable; and a driving means for rotating an intermediate portion of the cable at a predetermined rotation speed around the rotation axis, and the rotating body rotates at a predetermined rotation speed of 2 due to the anti-twisting force of the cable.
A continuous centrifugal separator that rotates at twice the rotational speed and has a structure in which the inlet conduit and the outlet conduit are connected to the blood container so as to be able to communicate with each other is more preferably used.

The blood introduction means in the present invention is a part that can be connected to a normal blood collection device or blood storage device such as a shunt or a syringe needle, and if necessary, it can be used in conjunction with other conduits, cots, pumps, etc. to transfer blood to white blood cells. It can be introduced into a concentration means. The treated blood lead-out means is a part that can be connected to a shunt, drip set, etc., and if necessary, it can be used in conjunction with other conduits, pots, pumps, blood pressure control valves, etc. to lead out the treated blood from the blood processing device. can do.

In carrying out the present invention, a buffer tank may be further provided in the circuits shown in FIGS. 1 and 2, or plasma purification means may be combined.

Next, the present invention will be described in more detail with reference to Examples.

Example 1 An IBM2997 continuous centrifugation device was used as a leukocyte concentration means, and a leukocyte separation means having the structure shown in Fig. 3 (separation material: acrylonitrile fiber 0.7 denier,
Blood processing was performed using the blood processing apparatus shown in FIG. 2 using pig blood 2 whose packing density was 0.16 g/cm ² ) and whose coagulation was prevented with ACD-A solution.

The number of red blood cells in pig blood before treatment was 5.7 million cells/μ,
White blood cell count is 18,000/μ, platelet count is 650,000/μ.
It was μ. The number of white blood cells after passing through the leukocyte separation means is 150 cells/μ, the number of red blood cells in the blood after passing through the mixing means is 5.7 million cells/μ, and the number of white blood cells is 860 cells/μ.
The number of platelets was 580,000/μm ^{3 .}

Comparative Example 1 When pig blood was directly passed through the leukocyte separation means of Example 1, the number of red blood cells in pig blood immediately after the distribution was
The number of white blood cells was 5.45 million cells/μ, the number of white blood cells was 530 cells/μ, and the number of platelets was 47,000 cells/μ. After approximately 800 ml of pig blood had passed through, the blood stopped flowing due to blood clog.

Example 2 The same treatment as in Example 1 was carried out, except that the blood treatment was carried out at 37°C using the apparatus shown in Figure 5 of Japanese Patent Application No. 16190/1983 as a leukocyte concentration means. However, the number of red blood cells in the blood after passing through the mixing means is 5.7 million cells/μ, the number of white blood cells is 1600 cells/μ,
The platelet count was 560,000/μ.

Example 3 As leukocyte separation means, polyester fibers (1.5 denier, packing density 0.10 g/cm ² ), polyacrylonitrile fibers (0.7 denier, packing density
Blood processing was carried out in the same manner as in Example 1, except that two separation means, each filled with 0.16 g/cm ² ), were used.

The number of white blood cells in the blood after passing through the polyester fiber-filled separation means is 8,700 cells/μ, and the number of white blood cells after passing through the polyacrylonitrile fiber-filled separation means is 57 cells/μ.
It was μ. After passing through the mixing means, the number of red blood cells in the blood was 5.6 million/μ, the number of white blood cells was 430/μ, and the number of platelets was 540,000/μ.

As described above in detail, by using the blood processing method and device of the present invention, it is possible to safely treat diseases related to immune reactions by treating leukocytes or lymphocytes without causing side effects such as promotion of bleeding tendency. The cells can be removed from the blood, making it possible to provide a new and extremely useful therapeutic means for diseases involving immune reactions, which have traditionally been considered intractable diseases.

[Brief explanation of drawings]

FIG. 1 is a schematic diagram showing an embodiment of the blood processing apparatus of the present invention, FIG. 2 is a schematic diagram showing another embodiment, and FIG.
The figure is a perspective view showing the configuration of the leukocyte separation means of the present invention. 1... Blood introduction means, 2... Leukocyte concentration means,
3... Leukocyte separation means, 4... Mixing means, 5...
Processed blood derivation means, 6...pump, 7...blood,
8... Red blood cell concentration component, 9... White blood cell concentration component,
10...Plasma component, 11...Processed leukocyte concentration component, 12...Processed blood, 13...Separation material, 14...
...Cylinder, 15...Ring, 16...Mesh, 1
7, 18...Silicone packing, 19...Leukocyte concentration component lead-in/out tube, 20...Lid, 21...Tightening ring.

Claims (1)

[Scope of Claims] 1. In a blood processing device for removing leukocyte components from blood, a leukocyte concentration means and a method for removing leukocyte components from the concentrated leukocyte concentration component are provided between the blood introduction means and the processed blood extraction means. 1. A blood processing device comprising leukocyte separation means. 2. A leukocyte concentration means provided between the blood introduction means and the processed blood extraction means is configured to separate blood into a leukocyte concentration component, a red blood cell concentration component, and a plasma component, and further separates the treated leukocyte concentration component from the red blood cell concentration component. 2. The blood processing apparatus according to claim 1, wherein a mixing means for mixing the components and the plasma component or its substitute is provided before the treated blood deriving means.