JPH02150274A - Suspension of immunological memory cell and preparation thereof - Google Patents

Abstract

PURPOSE:To prepare a cell suspension containing immunological memory cells in high density by effectively utilizing various characteristics and attributes of an immunological memory cell included in T-cell, thereby separating and purifying the cell in high efficiency. CONSTITUTION:A cell group is collected from a body immunized with a desired materies morbi and a cell cluster consisting essentially and exclusively of T-cell is separated from the cell group. An antibody specifically reactive with a desired histocompatibility antigen is added to the separated cluster and made to react with cell exhibiting positive surface antigen activity to the histocompatibility antigen among the cells of the cell cluster. The antigen-antibody complex formed by the reaction is bonded with a proper complement to break and remove the cell having positive antibody activity and exclusively leave an immunological memory cell group having negative histocompatibility antigen activity. Cells having surface antigen is separated and collected from an immunological memory cell group by reacting the immunological memory cell with an antibody against the surface antigen characteristic to said cell.

Description

[Detailed description of the invention]

[Industrial Field of Application 1] The present invention relates to a method for preparing an immune memory cell suspension, and in particular to a method for preparing an immune memory cell suspension that is effective for the prevention and treatment of infectious diseases and malignant tumors, and This relates to an immune memory cell suspension obtained by the preparation method. [Prior Art] Once a living body reacts to a certain type of antigen, it exhibits a strong reaction when it is next stimulated by the same type of antigen.
In the immune phenomenon of a living body, especially the acquired immune phenomenon, such an effect is known as immunological memory, and the above-mentioned immune reaction is called a secondary immune response or an anamnesis reaction. Such an immune phenomenon is caused by excessive proliferation of lymphoid cells capable of reacting to the antigen as a result of a response to the previous antigen stimulation. These cells are generally referred to as immunological memory cells, and are a type of lymphocyte contained in the lymph and blood circulating in the lymphatic system and vascular system of the living body. Lymphocytes migrate from the circulatory system into the thymus and combine with thymosin and 1
flil! ! mature by doing]7. They acquire immune ability and differentiate into so-called T cells (1972 cells). These T1111 cells are also called thymus-dependent cells or thymus-derived cells, and are composed of nine 1111! cells.
around the postcapillary veins in the paracortex of Iia and lymph nodes;
In addition to being particularly abundant in areas such as the central arteries of the NV and M white pulp, it is also known that they are present in almost all peripheral vessels of the circulatory system. 1. Immunological memory cells, which are a type of mature lymphocytes as described in 1-1, have so far only been conceptually recognized as immunological memory cells, as described above. The cells themselves, or at least the entities responsible for them, have not yet been isolated or extracted in a glass container, and such cells cannot be used to prevent or treat infectious diseases, malignant tumors, etc. There has never been a concrete prospect of putting it into practical use for this purpose, and no clinical or even experimental attempts have been made for that purpose. [Means for solving the problem] Not yet used. By effectively utilizing the various characteristics and attributes of immune memory cells contained in T cells involved in immune response, we are able to isolate and purify these cells with high efficiency to create a high density of immune memory cells. Preparing a suspension containing a
The object of the present invention is to provide a method for activating immunological memory cells, and a suspension of immune memory cells obtained thereby. In order to achieve this purpose, the invention has been developed to treat certain diseased individuals,
In other words, a cell population consisting essentially only of T cells from a cell group collected from an individual immunized against a specific virus, bacteria, or other pathogen, or cancer cells (in the present invention, these are collectively referred to as diseased individuals). Minute #17. An antibody that specifically reacts with a predetermined histocompatibility antigen is added to this cell population, and the histocompatibility antigen is reacted with cells in the cell population that exhibit positive surface antigen activity, and the antigen formed as a result of this reaction is By binding an appropriate complement to the antibody complex, the cells positive for the antigen activity are destroyed and removed, leaving only the immune memory cell group showing negative for the histocompatibility antigen activity, and thus obtained. An immunological memory cell group characterized by reacting an antibody against a surface antigen specific to the immune memory cell group, and separating and collecting cells having the surface antigen from the immune memory cell group! The present invention provides a method for preparing a suspension. In this W manufacturing method of an immune memory cell suspension, when separating and collecting cells having the surface antigen from the immune memory cell group, a layer of the anti-IgG antibody on the surface coated with the anti-IgG antibody is used. By causing the immune memory cell group to undergo a contact 2 reaction. It is preferable that cells having the surface antigen among the immune memory cell group are allowed to adhere to the surface, and then cells that are adherent to the surface are collected. When using a mouse as an individual, the predetermined histocompatibility antigen is the Ia antigen, and an anti-Ia monoclonal antibody that specifically reacts with this is added to the cell population to prepare the cell population. By forming an antigen-antibody complex with cells in which the Ia antigen activity is positive, and binding the appropriate complement to this, the cells that are positive in the Ia antigen activity are destroyed and removed, and the Ia antigen activity is reduced. It is possible to leave only negative immune memory cells and to react with the surface antigen by adding an antibody against it to these immune memory cells. In addition. As is well known, the Ia antigen in mice corresponds to the histocompatibility antigen HLA-R (or HLA-DR) in humans. Furthermore, the present invention provides for isolation of cells from a group of cells collected from individuals immunized against a predetermined pathogen. The present invention provides an immune memory cell suspension consisting essentially only of T cells, characterized in that approximately 58% of these T cells contain at least 1×10 3 immune memory cells.

[Effect]

The immune memory cell suspension according to the present invention can theoretically be isolated and purified from the thorax-dependent region of the circulatory system of individuals who have healed from all currently known infectious diseases and malignant tumors. Therefore, as long as individuals with a history of healing are available, they exert an immune effect against any bacterial or viral infection or malignant tumor. Bacterial and viral infections are caused by two types of bacteria, such as Salmonella and Carnebacterium.
, Pseudomonas , Pasteurella Streptococcus , Ephthromelia virus
lia virus), Sendai virus (HVJ)
) and other pathogenic microorganisms are typical. Furthermore, the immune memory cell suspension according to the present invention. This is because the immune mechanism resulting from this is fundamentally different from the immune mechanism based on antigen-antibody reactions. The conventional methods of preventing infectious diseases that use vaccines as antigens do not cause side effects.
It also has the advantage of being completely impossible theoretically. [General Aspect of the Invention J In order to prepare an immune memory cell suspension by the method described above, the present invention first involves immunizing cells against a suitable cancer or some bacterial or viral infection. Cell groups containing lymphocytes are isolated from individuals with a history of disease (hereinafter simply referred to as cured individuals). This cell group containing lymphocytes can be extracted from the peripheral blood collected from the heart or vein of the cured individual, or dissociated from the spleen tissue of the individual, and then start with either of these cell groups. It is easy to use as a material. In the following description, these peripheral blood or spleen tissues will be used as starting materials. From this lymphocyte-containing cell group separated from peripheral blood or spleen tissue as a starting material, T cells are collected into the cell suspension by column chromatography or the like using a solid phase adsorbent as a separation means to selectively elute T cells. Cells such as B cells, which are responsible for humoral immunity, are removed from the cell group contained therein, and a suspension containing a cell population consisting only of T cells is fractionated and eluted. Nylon wool is preferably used as the solid phase adsorbent used in this column chromatography process. However, the solid-phase adsorbent such as Nylon Cool adsorbs not only B cells but also macrophages, white blood cells, etc.
The adsorption efficiency of B cells themselves becomes relatively low. Therefore, in order to increase the adsorption efficiency of B cells to this nylon wool. As is known, prior to column chromatography using nylon wool, column chromatography using another solid phase adsorbent, preferably glass wool, is preferably performed. Both T cells and B cells have low adhesion to the solid-phase adsorbent made of glass wool, but macrophages and leukocytes show high adhesion; therefore,
By performing column chromatography treatment using this glass wool, macrophages and leukocytes are adsorbed and removed in advance, and then by column chromatography treatment using nylon wool, Tm cells and B cells can be separated with high efficiency. It becomes usable to do this. However, in addition to column chromatography using a solid-phase adsorbent as described above, the separation of T cells and B cells can also be performed using methods that utilize the differences in surface antigens specific to these cells. In this case, when a mouse is used as a cured individual as described above, T cells and B cells can be separated using thyt antigen as a surface antigen for identifying T cells. The cell population thus obtained is mostly composed of T cells as described above, but it also contains a small amount of immature lymphocytes that lack immunological ability. Therefore, in the next treatment, these immature lymphocytes are removed by causing the cell population to experience an antigen-antibody reaction using the cell population as an antigen. To do this, first, an antibody that specifically reacts with a predetermined histocompatibility antigen is added to the cell population, and the antibody is reacted with cells in the cell population that exhibit positive surface antigen activity for the histocompatibility antigen. By binding complement consisting of an appropriate serum (typically rabbit serum) to the antigen-antibody complex formed as a result of this antigen-antibody reaction. Lymphocytes positive for the antigen activity are destroyed and removed, leaving only immune memory cells negative for the histocompatibility antigen activity. When using a mouse as a cured individual, the predetermined histocompatibility antigen is Ia
An anti-Ia monoclonal antibody that specifically reacts with the Ia antigen is added to the cell population to form an antigen-antibody complex with cells positive for Ia antigen activity in the cell population. By fixing complement with the above-mentioned appropriate serum, lymphocytes positive for Ia antigen activity are destroyed and removed.
Ensure that only immune memory cells negative for antigen activity remain. As mentioned above, the Ia antigen in mice is the histocompatibility antigen HLA-R (or HLA-DR) in humans.
This corresponds to The cells obtained as a result of the above-mentioned antigen-antibody reaction and complement fixation reaction are reacted with a surface antigen specific to the immune memory cells by adding, for example, a rat/rat antibody that opposes the surface antigen as an antigen. In addition to immune memory cells that have been subjected to such treatment, in which the surface antigen is coated with antibodies,
anti-depressants using appropriate anti-depressant IgG serum as a starting material)
An IgG antibody is prepared, and a cell suspension containing the above immune memory cells is placed in a glass vessel whose inner surface is coated with the anti-IgG antibody and allowed to react. As a result, the anti-cat IgG antibody acts as an antibody against the rat antibody covering the surface antigen, that is, as an antigen in the reaction system, and an antigen-antibody reaction occurs between the anti-cat IgG antibody and the rat antibody. To occur. The cells coated with the rat antibody will adhere to the inner surface of the glassware. However. Cells non-adherent to the inner surface of this glassware. That is, after removing the cells that do not have the surface antigen, only the cells that are adherent to the inner surface of the glass are collected, and a cell suspension containing these cells is used as the immune memory cell according to the present invention. Make a suspension. In addition, the surface antigens of the immune memory cells include Thyl antigen, l,! tl antigen, Lrt2 antigen, L,! T3 antigen and the like are known. This immune memory cell suspension according to the present invention corresponds to approximately 0.1% of the Ta cells physically separated as described above. As will be clear from what will be described later, from the results of various experiments conducted by the present inventor, the majority of these cells, approximately 58% of the cells, are
When mice are used as the cured individuals, it has been confirmed that immunological memory transfer will occur with a substantially 100% probability if the number of these cells is at least 1×103. "Effect" The immune memory cell suspension according to the present invention prepared as described above causes cells to express an antigen-specific immune mechanism, that is, a cellular immune mechanism, and is capable of producing immunoglobulins and monoclonal antibodies. This is fundamentally different from the mechanism of immunity based on the antigen-antibody reaction (humoral immune reaction), which is controlled by B cells and contributes to the immune system. Therefore, clinically, the immune memory cell suspension according to the present invention is completely unlikely to cause any side effects associated with, for example, conventional infectious disease prevention methods using vaccines as antigens. The immune effects obtained by immune memory cell suspensions can theoretically be used to prevent or treat all known bacterial and viral infections and malignant tumors, as long as cured individuals are available. It is effective against [Example] Next, specific examples of the method for preparing an immune memory cell suspension according to the present invention will be described.

[1] Isolation of immune memory cells In this example, a mouse with a history of being cured from cancer or a bacterial or viral infection was used as the cured individual, and peripheral blood or The lung tissue was collected. <<t-i) Preparation of cell suspension■ When using peripheral blood as a starting material, no pretreatment procedure is required, and the collected peripheral blood is immediately diluted with 5 times the amount of Hanks. Suspended in pressurized liquid. This mixture was centrifuged at 4°C for 15 minutes at 9 revolutions and 2000 rP to extract the white blood layer (buffy (coat) was gently sucked up and separated. ■This white blood layer fraction was similarly suspended in 5 times the volume of Hank's eluate, and then centrifuged at 4°C for 15 minutes at 9 rotations of 200 rpm in the same manner as the above centrifugation.
The supernatant was removed. The cell suspension thus obtained was further centrifuged twice under the same conditions as above, and the supernatant was removed each time. The cell group obtained by washing by centrifugation three times was placed in Hank's solution containing 5% fetal bovine serum (FSC) at a cell density of 4.
This was used as the "initial J cell suspension" in this example. ■On the other hand, when lung tissue is used as a starting material, first the cured individual mouse After cutting out a part of the lung tissue, the cut tissue pieces were placed in a stainless steel strainer (mesh number = ↑yler standard screen scale No. 1).
00), and the cells that passed through the mesh pores were added to an appropriate amount of Hank's eluate and suspended. For the suspension containing lung cells mechanically dissociated in this way, 4
℃ for 15 minutes. Centrifuged at a rotational speed of 2000 rp,■
Thereafter, the same centrifugation operation as in the washing process described in (1) above was performed three times, and Hank's eluate containing 5% fetal bovine serum was added to the cell population thus obtained to a cell density of 4 x 107 cells. In addition, this was used as the "initial" cell suspension in this example. 1-2) Isolation of T cells ■ The initial cell suspension obtained from peripheral blood or lung tissue as described above was first transferred to a glass tube (inner diameter 2cm high, length 10mm).
cm) was infiltrated into a glass wool column packed with a glass wool column column, and Hanks' eluate containing 5% fetal bovine serum was layered on the top of the column at 37°C for 4 hours.
Incubated for 5 minutes. Thereafter, while maintaining the temperature of the column at 37° C. and gently adding the above solution to the column, the cock at the bottom of the glass tube was opened, and the cell suspension eluted in 9 fractions was collected in a centrifuge tube. As mentioned above, T cells and B
All cells have low adhesion to glass walls,
Therefore, by column chromatography treatment using glass wool, cells other than T cells and B cells, particularly macrophages and leukocytes, selectively remain attached to the glass wool. Therefore, the cell group contained in the cell suspension separately eluted from the glass wool column by the two-tube treatment consists essentially of only T cells and B cells. The cell suspension thus collected contains approximately 30% of the cell population in the initial cell suspension initially added to the glass wool column. ■Next, the cell suspension collected in the centrifuge tube was heated to 4°C.
After centrifugation at 9 rotations and 200 rpm for 15 minutes, the supernatant was removed. The thus obtained cell population was added to Hank's Pressure Solution containing 5% fetal bovine serum until the cell density was 4 x 10.
Suspend the cells so that 7 cells/cover are obtained, and transfer this cell suspension to a glass tube (inner diameter 2C, length 10C) with nylon wool.
0 g (liB+11114 borataries) packed with nylon wool column,
The column was incubated again at 37° C. for 45 minutes with Hank's pressure solution containing % fetal bovine serum layered on top of the column. Thereafter, as in case ① above, while maintaining the temperature at 37°C, the above solution was gently added to the column, the cock at the bottom of the glass tube was opened, and the cell suspension eluted in 9 fractions was collected. This eluted cell suspension contains approximately 33% of the cells in the cell suspension added to the nylon wool column.
%, and therefore approximately 10% of the cell population initially added to the glass wool column was recovered. Most of the cell groups obtained by the two-step column chromatography treatment as described above, that is, approximately 85% according to the experimental results, are T cells.9 The cell groups unnecessary in the present invention are T cells. Most of it was adsorbed by the fibers in the glass wool column or nylon wool column and remained in the column. However, as mentioned above. The cell group at this stage also contains a small amount of unnecessary lymphocytes. (1-3>> Purification of T cells ■ In order to remove these unnecessary lymphocytes, etc., the cell QsJ solution fractionated and eluted by the nylon wool column chromatography treatment was first rotated at 4°C for 15 minutes at 1 rotation speed. Centrifugation was performed at 200 rpm.The supernatant was removed from the cell suspension obtained by this centrifugation, and the resulting cell group was placed in 199 medium containing 2% fetal bovine serum.
The cells were suspended at a density of 8 x 107 cells/mayumi. Next. For one volume of this cell suspension, the same medium as above (2%
Anti-Ia monoclonal antibody (Cedarlane Laboratories) diluted 30 times with 199 medium containing fetal bovine serum (the same applies hereinafter)
ries) and the same medium as above.
Add 1 volume of double diluted rabbit serum and incubate for 30 minutes at 37°C.
Allowed to react for minutes. As a result, Ia in the cell suspension
An antigen-antibody complex is formed between cells positive for antigen activity and the anti-Ia monoclonal antibody, and complement from rabbit serum binds to this antigen-antibody complex, and cells positive for Ia antigen activity are destroyed and removed. Only immune memory cells negative for Ia antigen activity will remain. The cell suspension thus obtained was centrifuged at 4° C. for 15 minutes at a rotation speed of 200° rpm to collect the cells. ■After centrifuging these cells three times under the same conditions and using the same medium as above, the cell density becomes 8 x 107 cells in the same medium as above. It was suspended like this. (2) Separately from this cell suspension, a rat antibody was prepared by the method described below, and diluted 5 times with the same medium as above. The rat antibody and the cell suspension were added in equal amounts, and reacted at 4°C for 60 minutes. With this process,
Rat antibodies, which are antibodies against antigens specific to immune memory cells, bind to and coat the antigens. A cell suspension containing an immune memory cell group having such an antigen-antibody complex was heated at 4°C for 15 minutes at 2,000 revolutions.
Cells were collected by centrifugation as rp■. These cell groups were further centrifuged three times using the same medium as above under the same conditions, and then the same medium as above was added so that the cell density was 8×IO6 cells/layer. suspended in. ■Furthermore, separately from this cell suspension, anti-cat IgG goat serum was diluted 30 times with the same medium as above, and the diluted 10 times
Plastic petri dish (inner diameter 10cm, Becton Dickinson)
(manufactured by Company N) and left it at 4°C overnight. Excess antibody was removed by washing, and the anti-cat IgG goat antibody was uniformly adhered and remained on the inner surface of the petri dish. In this way, 10 mounds of the above cell suspension were placed in a petri dish on which a liquid film of IgG goat antibody was formed on the inner surface and gently held horizontally at 4°C for 60 minutes. (2) Thereafter, the petri dish was gently shaken to remove cells that had relatively low adhesion to the liquid film on the inner surface of the petri dish, ie, cells that did not have the antigen specific to the immune memory cells. R containing 10% fetal bovine serum on the inner surface of this petri dish.
The relatively highly adherent cells were collected by washing three times with PMI-1640 medium and then strongly spraying the medium against the inner surface of the petri dish. An immune memory cell suspension was prepared. The cells contained in the cell suspension prepared by the above operations ■ to ■, that is, the immune memory cells, are the cells contained in the cell suspension prepared from the above operations, that is, the cells contained in the cell suspension eluted from the nylon wool column. This corresponds to approximately 0.1% of the cell group consisting mostly of T cells. This 0
．． Results of various experiments conducted by the present inventor on 1% of cells. Approximately 58% of these cells were confirmed to have the following characteristics. Belongs to small spherical lymphocytes. ■Shows positive antigen activity for memory cell-specific antigen. ■ThY1 antigen, Lytl antigen, Lyt2 antigen,
The antigenic activity was positive for the t4t3 antigen, and negative for the Ia antigen. ■When this immune memory cell suspension is injected into a mouse of the same strain as the mouse used as a cured individual in this example, the immune memory cells in the immune memory cell suspension are at least I
If 103 are included, there is a virtually 100% probability that immune memory will be transferred. ■If an individual transplanted with the immune memory cells is irradiated with cobalt gamma rays, it will not lose its memory even if it is irradiated with 600 roentgens, but will lose its memory if it is irradiated with 1100 roentgens. This indicates that the cells belong to a cell group that has considerable resistance to radiation.

[2] Preventive effect of immune memory cells on disease An individual transplanted with the immune memory cells contained in the immune memory cell suspension of the present invention isolated and purified as described above (transplanted individual) was infected with the disease than the initially cured individual. The immune memory cells were challenged with pathogenic microorganisms or cancer cells of the same species as the diseased individual, and the disease prevention effect of the immune memory cells was evaluated. A mouse was used as the healing body. In addition, the diseased individuals include cancer cells, Salmonella enterica, Corynebacterium, Pseudomonas bacterium, and Pasteurella bacterium. Streptococcus, Ectromelia Φ virus, and Sendai virus (HVJ) were used. The table below shows the results. mouse power

[Cancer cells]

LJL Agriculture JLJ 4 IXIQ l　X　103 pieces disease E-U-pot 100% 100% 6 x 102 pieces 3 x lO2 pieces l　X　102 pieces

[Salmonella]

1” L cliff JLJ IX 104 pieces IXIQ” pieces 6 x 102 pieces 3 x 102 pieces IX 102 pieces

[Coryneba Clear]

Vines - one plant, one plant, one plant 4 IXIQ 3 IXIQ 6 x 102 pieces 3 x 102 pieces 1 x 102 pieces

[Pseudomonas]

1jL Cliff” Ll 4 IXIQ 70% 42% 34% E-” missing-” L-ri 100% 100% 68% 34% 20% ヱJLIL-釆 100% 100% 48% 26% 14% 1-” Beg-” Shi-! 100% 1 x 103 pieces 100% 6 x 100 pieces 41% 3 x 102 pieces 23% lX102 pieces 11%

[Basturella bacteria]

Affairs-Suichi 1-4-Dou Sat-JLJL-Kan txio
'1 piece 100% 1 x 103 pieces 100% 6 x 102 pieces 78% 3 x 100 pieces 53% 1 x 102 pieces 35%

[Streptococcus]

Vines - 1 - follicles - fi 1-1L" 1-jlX
104 pieces 100% I X 103 pieces 100% 6 X 102 pieces 44% 3

[Eftromelia virus]

1-JL”9-】100% 100% 51% 27% 13% 1 x 104 pieces 1 x 103 pieces 6 x 102 pieces 3 x 102 pieces 1 x 102 pieces

[Sendai virus]

Fu-Shi-nicho-Emperor IJLj Kai-''lX104
I [9100% 1X103 pieces 100% 6 x 102 pieces 88% 3
63% l x 102 cells 29% As is clear from the above experimental results, for any of the above diseased individuals, immune memory cells derived from cured individuals were
If 103 or more immune memory cells are transplanted, the individual who has not had the immune memory cells transplanted (non-transplanted individual) will be infected with an amount of bacterial or viral pathogen that has a 100% probability of becoming ill, or the non-transplanted individual It has been experimentally confirmed in mice that even if a patient is transplanted with an amount of cancer cells that causes a 100% probability of becoming infected, it is possible to completely suppress the onset of disease caused by pathogenic microorganisms or cancer cells. Furthermore, as is clear from the above experimental results, when mice are treated as cured individuals, the number of transplanted immune memory cells is l x 1
Even if the number of immune memory cells is less than 03, although it differs slightly depending on the target disease individual, there is approximately a 50% probability that by transplanting 6×102 ([1) immune memory cells, and 3×102 immune memory cells will also be transplanted. It has also been experimentally confirmed that by doing so, it is possible to prevent the onset of diseases caused by the respective disease individuals with a probability of approximately 30%. [3] Preparation of anti-memory cell rat antibodies The rat antibody used in the treatment of (1-3)■ was prepared as follows. First, the immune memory cells collected as described above (1-:l■) were phosphate-buffered. The suspension was suspended in physiological saline, and the suspension was subjected to normal centrifugation under appropriate conditions.These operations were repeated three times to wash the resulting cells. The cell population was suspended in the above-mentioned phosphate buffered saline at a cell density of 1×10” cells/an. 0.25 ml of the suspension thus obtained was added to a Wistar female rat. Furthermore, on the 10th and 20th day from the day of this first administration, the same number of cells was administered subcutaneously to the thigh of the female rat. On the 30th and 40th days from the day when the above-mentioned phosphoric acid Suspended in buffered saline, its 0.2
5mM was administered intraperitoneally to the rats. Further, on the 50th to 55th day from the day of the first administration, whole blood was collected from the rats. Thus, 5 mi to 6 ml of serum was obtained from one rat. Separately from the above-mentioned treatments, a portion of lung tissue was excised from a newly born mouse that was syngeneic to the mouse from which the immune memory cells were collected. This cut tissue piece was strongly pressed against a stainless steel strainer similar to that described above to collect cells that had passed through the mesh pores. WII mechanically dissociated in this way
1%! The cells were suspended in the phosphate buffered saline, and the resulting cell suspension was centrifuged. Repeat this process three times, add about 1.5 times the amount of rat serum to the final cell mass, stir thoroughly, and then leave it in water for 60 minutes. The mixture was further stirred thoroughly. Thereafter, after 60 minutes had elapsed, the suspension was subjected to normal centrifugation under appropriate conditions, and the supernatant thus obtained was subjected to the same operations as above. This treatment was repeated three times, and the supernatant finally obtained was used as the target anti-memory cell rat antibody in the treatment described in ((1-3)) (2) above. [Effects of the Invention] The immune memory cell suspension according to the present invention can be isolated, purified, and cultured from the thymus-dependent region of the circulatory system of individuals with a history of healing from any currently known infectious disease or malignant tumor. Therefore, as long as individuals with a history of healing are available, they exhibit an immune effect against any bacterial or viral infection or malignant tumor. Furthermore, since the immunity mechanism of the immune memory cell suspension according to the present invention is fundamentally different from the immunity mechanism based on antigen-antibody reactions, infection using a conventional vaccine as an antigen is not possible. It is completely impossible for the method to prevent the disease to cause side effects, etc. Although two embodiments of the present invention have been described above, the method according to the present invention may be carried out by adding or changing the described embodiments as appropriate. Needless to say. Applicant Geo Research Co., Ltd.

Claims (3)

[Claims] (1) Separating a cell population consisting essentially only of T cells from a cell population collected from an individual immunized against a predetermined pathogen, and reacting specifically with a predetermined histocompatibility antigen with this cell population adding an antibody to react with cells in the cell population showing positive surface antigen activity for the histocompatibility antigen;
By binding appropriate complement to the antigen-antibody complex formed as a result of this reaction, only the immune memory cell group showing negative histocompatibility antigen activity remains, and the thus obtained immune memory cell group A method for preparing an immune memory cell suspension, which comprises reacting an antibody against a surface antigen specific to the cells, and separating and collecting cells having the surface antigen from the immune memory cell group. (2) When separating and collecting cells having the surface antigen among the immune memory cell group, the immune memory cell group is brought into contact with the anti-IgG antibody layer on the surface coated with the anti-IgG antibody and reacted with it. By attaching cells having the surface antigen among the immune memory cell group to the surface,
The method for preparing an immune memory cell suspension according to claim 1, wherein cells adherent to this surface are then collected. (3) Isolated from a population of cells collected from an individual immunized against a given pathogenic agent and consisting essentially only of T cells, approximately 58% of which are at least 1 x 10^ An immune memory cell suspension characterized by containing three immune memory cells.