WO2003012080A2

WO2003012080A2 - Ex-vivo selection of t-cells with reduced alloreactivity and use thereof in bone marrow transplantations

Info

Publication number: WO2003012080A2
Application number: PCT/EP2002/008052
Authority: WO
Inventors: Ahmed Sheriff
Original assignee: Genethor Gmbh
Priority date: 2001-07-21
Filing date: 2002-07-19
Publication date: 2003-02-13
Also published as: WO2003012080A3; AU2002321240A1

Abstract

The invention relates to a reduction of allologous immune reactions. In said method the regulatory properties of antigen-presenting cells are used to eliminate ex-vivo graft vs host reactions.

Description

Ex vivo selection of T cells

The present invention relates to antigen-presenting cells, methods for producing antigen-presenting cells, medicaments containing antigen-presenting cells and use of the antigen-presenting cells.

description

For almost 10 years, gene therapy methods have been developed and used for various diseases and animal products. So far, they have not become routine. Most of the time, this is the treatment of genetically determined, serious diseases for which other therapies are not available. Furthermore, gene therapies are used to treat severe and also non-treatable cancer.

General information about transplants

Tissue transplantation to replace diseased organs is an important medical therapy today. In most cases, an adaptive immune system response to the graft poses the greatest threat to successful treatment. Adaptive immune system responses are induced by antigen presenting cells by activating T helper lymphocytes. When transfusing blood, which is the first and most commonly used graft, the ABO and Rh blood group antigens must be matched to avoid the rapid destruction of inappropriate erythrocytes. For other tissues, the very polymorphic major histocompatibility complexes (Tissue Compatibility Complexes, MHC) can be compared to each other, since these almost always trigger the immune response. Unfortunately, perfect matching of the MHCs is almost impossible except for relatives, since the degree of agreement in the MHC pattern is often too low. Stem cell transplants for leukemia

Leukemia is cancer of the blood cells. 50 out of 1 million people get leukemia every year. When leukemia develops, the body produces large amounts of abnormal blood cells. In most types of leukemia, the abnormal cells are white blood cells.

leukemias

There are several types of leukemia, which essentially show two characteristics. One characteristic is how quickly the disease develops and worsens. The other is determined by the type of blood cell that is affected.

Leukemia is either acute or chronic. In acute leukemia, the abnormal blood cells are very immature blasts that cannot perform their normal functions. The number of blasts increases rapidly and the ailments quickly get worse. Some blasts are present in chronic leukemia, but generally these cells are more mature and can perform some of their normal functions. The number of blasts also rises more slowly than with acute leukemia. With chronic leukemia, the disease gradually gets worse.

Leukemia essentially appears in two main types of white blood cells, lymphoid cells or myeloid cells. Accordingly, leukemia occurs as lymphatic leukemia or _. myeloid leukemia. The most common leukemias are:

- Acute lymphoblastic leukemia (ALL) is the most common leukemia in young children. This disease can also occur in adults, especially from the age of 65.

- Acute myeloid leukemia (AML) occurs in adults and children. This type of leukemia is sometimes called acute non-lymphoblastic leukemia (ANLL). - Chronic lymphoblastic leukemia (CLL) mostly affects adults over the age of 55. It sometimes occurs in younger adults, but hardly ever affects children.

- Chronic myeloid leukemia (CML) mostly occurs in adults. A small number of children also develop this cancer.

Symptoms of leukemia

Leukaemic cells are abnormal cells that cannot function as normal blood cells. They cannot help the body fight infections. Because of this, people with leukemia often develop infections and have a fever.

People with leukemia often also have less healthy red blood cells and platelets, so there are not enough red blood cells available to transport oxygen through the body. Due to anemia, patients are often pale and feel dull and tired. If the number of platelets is too low, patients tend to bleed easily and quickly develop blue hematomas.

Like all blood cells, leukemic cells circulate in the body. Due to the number of abnormal cells and their location, patients with leukemia can show various symptoms. In acute leukemia, symptoms appear quickly and the patient's condition also deteriorates rapidly. Chronic leukemia appears

symptoms for a long time. When symptoms appear, they are usually mild at first and the patient's general condition only gradually worsens.

Symptoms for leukemia:

* Fever, chills, and other flu-like symptoms;

* Weakness and fatigue;

* frequent infections;

* Loss of appetite and / or weight; * swollen or sensitive lymph nodes, liver, or spleen;

* rapid bleeding or bruising;

* small red spots under the skin;

* swollen or bleeding gums;

* Sweating, especially at night; and / or * bone or joint pain.

In acute leukemia, the abnormal cells can collect in the brain or spinal cord (central nervous system or CNS). The result can be headache, nausea, nausea, confusion, loss of muscle control and strokes. Leukaemic cells can also collect in the testes and cause swelling there. Some patients develop painful eyes or skin. Leukemia can also affect the digestive tract, kidneys, lungs, or other parts of the body.

With chronic leukemia, the abnormal cells can gradually collect in different parts of the body. Chronic leukemia can affect the skin, CNS, digestive tract, kidneys and testicles. Treatment of leukemia:

Treating leukemia is complex. It varies with the type of leukemia and is not the same in all patients. Treatment also depends on

certain characteristics of the leukemic cells, the extent of the disease, whether the leukemia has been treated before. The patient's age, symptoms and general health are also relevant factors.

Acute leukemia must be treated immediately. Patients with chronic leukemia who show no symptoms do not need to be treated immediately. Unfortunately, chronic leukemia can rarely be cured.

Treatment methods:

Most patients with leukemia are treated with chemotherapy. Some are additionally irradiated and / or receive a bone marrow transplant (KMT, English: bone marrow transplantation = BMT) or biological therapies. In some cases, removal of the spleen can help. Before bone marrow transplantation, patients receive full body radiation combined with chemotherapy to destroy the leukemia-producing bone marrow.

The healthy bone marrow can come from a donor or it can come from the patient. It is then removed and treated outside the body to remove leukemic cells before high dose treatment. After that, a hospital stay of several weeks is necessary so that the transplant grows and enough white blood cells (leukocytes), such as T, B lymphocytes, neutrophils, basophils and eosinophils, platelets (platelets), monocytes and other subpopulations for the reconstitution (restoration) of the hematopoietic hematopoietic) system can produce. In the meantime, patients need to be protected from infection. The biological therapies include treatments with substances that influence the immune response to cancer. Cytokines, such as especially interferons, interleukins and colony stimulating factors (CSF) are used for example

some types of leukemia. They are usually combined with chemotherapy or bone marrow transplantation.

Bone marrow transplant

Bone marrow transplant patients have an increased risk of infection, bleeding, and other side effects from high doses of the chemotherapy drugs and radiation they receive. Graft versus host disease (GVHD) responses may also occur. The liver, skin and digestive tract are the organs most frequently affected. GVHD can be mild or severe and can occur at any time after the transplant (even years later). Immunosuppressants are given to reduce and treat the risk of GVHD.

Chronic myeloid leukemia

The mode of action of bone marrow transplants is explained below using the example of chronic myeloid leukemia: Allologic bone marrow transplantations (BMT) have provided treatment options for chronic myeloid leukemia (CML) in the past 20-30 years. However, no suitable donors can be found for about 60% of the patients.

CML is characterized in the early chronic phase by a single transformative genetic abnormality identified. It is the t (9; 22) translocation (Philadelphia Chromosome, Ph) that the bcr-abl oncogene creates, the only factor that is essential for the development of the Disease is needed (Daley et al 1990). Compared to other tumor types in the chronic phase, CML patients have a relatively intact immune system (Lewalle et al 1996). CML is accessible for a tumor-specific immune response. In the past 20-30 years, anti-tumor

Answers from allologic bone marrow transplants (allo-BMT) and recently also from donor leukocyte infusions [Donor leukocyte infusion (DLI)] clinically used for the treatment of CML.

The detection and extinction of remaining tumor cells by donor immune cells seems to be essential for the induction of a molecular remission. Transplants from which the T cells have been removed increase the risk of relapse to CML (Champlin et al 1988; Horowitz et al 1990). A stringent demonstration of the anti-leukemia effect generated by allo-BMT can be seen when DLI is administered as soon as the disease reappears. In this constellation, DLI can restore sustained molecular remission in up to 70% of cases (MacKinnon 2000). However, DLI is also associated with significant toxicity caused by GVHD. This often accompanies the graft-against-leukemia (GVL) effect. The mortality rate is 50 - 90% (MacKinnon 2000).

Instructions for immune regulation in CML:

- Increased risk of relapse in the event of T-cell exclusion from the graft. - Increased risk of relapse in the absence of GvHD

- BMT from syngeneic twins is less effective than matching BMT from siblings

- The relapse reacts to the discontinuation of immunosuppression

- Donor leukocyte infusions are effective in relapse.

Allo-BMT is a fairly rough approach with significant graft-related morbidity and mortality. The risk of dying is 20-41% (Silver et al 1999). This form of immunotherapy also offers egg ne up to 70% leukemia-free survival rate for the transplant recipients (Clift &. Anasetti 1997). This GvHD overlays the desired anti-leukemic effect.

Bone marrow transplants are also used for other leukemias.

Graft-versus-host disease (GvHD)

Graffc versus host disease (GvHD) is a major cause of morbidity and mortality after allogeneic transplantation and is caused by donor T-lymphocytes in the transplant. In HLA-compatible transplants, the GvHD response is directed against minor histocompatibility antigens. The recipient's APC presumably present the foreign tissue antigens to the donor T lymphocytes, in particular since the activity of the APC is increased by chemo- and radiotherapy-induced cytokines such as interleukin (IL) -1, IL-6 and TNFa. GvHD mainly affects the skin (small-spot rash), liver (cholestatic hepatitis) and intestine (diarrhea). In addition, mucous membranes and conjunctiva (Sicca syndrome) dry out and myositis. To prevent GvHD, immunosuppression is carried out with cyclosporin, possibly in combination with methotrexate and corticosteroids.

T cell depletion and graft versus leukemia effect

Depletion of T lymphocytes in the graft reduces the risk of GvHD. At the same time, however, the risk of tumor recurrence increases because the donor T lymphocytes also mediate the favorable graft versus leukemia effect (GvL). If you want to maintain the GvL effect while reducing the GvHD risk, you have to partially deplete the T cells and gradually reinfuse the donor T lymphocytes at later times. _ g.

Fas and trail

FasL is known as a potent apoptosis-inducing molecule (membrane protein). It communicates its effects via Fas, a receptor that occurs on T cells. If DCs are transfected with FasL, they induce apoptosis in

T cells (Matsue et al 1999). FasL-transfected DCs can cause an Ag-specific reduction in graft rejection. This is based on the interaction (apoposis induction) of FasL on DCs and Fas receptor on the T cells (Min et al 2000).

TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) is a recently identified member of the tumor necrosis factor cytokine superfamily. TRAIL induces apoptosis in various tumor cell lines. It is also suspected that TRAIL plays a role in the death of lymphocytes (Simon et al 2001; Miura et al 2001).

Co-stimulatory molecules

Functional, mature DCs arise from circulating progenitor cells after a maturation period. DCs can be characterized by a specific population of marker molecules. These include accessory, costimulatory gene products such as CD40, CD80, and CD86 as well as MHC class I and II. The CD83 molecule in particular is a well-characterized marker for fully mature DCs, since CD83 cannot be detected on immature DC precursors. However, the functional significance of CD83 is unknown. Its increasing presence during the ripening indicates an important function (Berchtold et al 1999).

Herpes simplex virus is able to suppress the expression of CD83 in an unexplained manner, while the production of other molecules typical of DCs such as CD25, CD40, CD80, CD86, CD95 and MHC of class I and class II remains unaffected (Kruse et al 2000b). Interestingly, Inhibition of CD83 expression on the plasma membrane leads to a dramatic reduction in DC-mediated T cell stimulation. If the eukaryotic initiation factor 5A is hindered by a specific inhibitor, CD83 expression can be prevented, which also leads to a strong reduction in DC-mediated T cell stimulation (Kruse et al 2000a). eIF-5A is a

Protein that is included in the export path of specific RNAs from the cell nucleus.

eIF-5A is the only cellular protein known to contain the unusual amino acid hypusin. This modification appears to be necessary for cell division. Hypusin modification is a spermidine-dependent post-translational reaction that is catalyzed by two enzymes. This includes the transfer of the aminobutyl group of spermidine to the ε-NH ₂ group of lysine at position 50 in eIF-5A by the deoxyhypusin synthase (Kruse et al 2000a).

The intermediate that is generated is then hydroxylated by the deoxyhypusin hydroxylase. In this way the active form of eIF-5A is created. Although eIF-5A was originally referred to as an "initiation factor", recent in vitro and in vivo experiments have shown that eIF-5A is not an initiator of protein translation. eIF-5A appears to be part of a specific export path from the cell nucleus, which is used, for example, by the Rev / Rex class of retroviral RNA transport factors. It was also shown that the eIF-5A protein accumulates on the nucleoplasmic side of the nuclear pore complex in order to interact with the general nuclear export receptor CRM1 and to migrate from the nucleus into the cytoplasm in mammalian cells. Studies of the amount of eIF-5A mRNA in human cells showed that elF-5A is constitutively expressed in cell lines as well as various other tissues. In contrast, the eIF-5A gene is induced in primary lymphoid cells during the activation and / or proliferation of primary human blood cells. If an inhibitor of hypusin modification [GC7 (N ¹ - guanyl-l, 7-diaminoheptan)], the expression of CD83 and thus the full stimulatory activity of mature DCs is inhibited (Kruse et al 2000a).

The cell according to the invention is preferably a dendritic cell or another antigen-presenting cell or one of the precursors of such a cell.

The APCs are the switching points of the adaptable immune system. Only they can trigger or stop a T cell-mediated immune response.

The allologic immune reactions - cannot be maintained without the help and mediation of APCs and T-helper lymphocytes. The cytotoxic T cells are also activated by APCs. They mediate a cellular immune response in which cells are killed directly by the cytotoxic T cells.

The molecular mechanisms of interaction - cell-cell contact - of APCs with the T helper and cytotoxic T lymphocytes are known in detail at the receptor level.

If the antigen is e.g. a bacterial protein, the immune response is useful for the organism. However, if the antigen is a transplant antigen, one speaks of a (pathological) rejection reaction.

In addition to the antigen to be presented (against which the antibodies to be produced are directed), various receptors and auxiliary receptors on the cell surface are involved in cell-cell contact and cell activation. An essential molecule of the intercellular interaction in the antigen presentation (contact of APCs with the T-lymphocytes) are costimulatory receptors with the designation B7 (CD80, CD86). WO 01/29192 discloses a method of using antigen-presenting cells to prevent GvHD effects. However, the antigen-presenting cells are co-incubated with dead or dying cell parts of the transplant recipient in order to contain loaded antigen-presenting cells

To get cells. The cell components should contain antigens for which a reduction in an immune response is desired. According to the invention, this step is not required since cells of the receiver are used. In addition, further changes are made or immature DCs are used to reduce a repulsive immune response.

The problem on which the invention is based is, inter alia, to make available therapeutically usable products for the reduction of allologic immune reactions in bone marrow transplants.

Figure 1 shows the recovery of non-alloreactive T cells with mature DC. Figure 2 shows the extraction of non-alloreactive T cells with immature DC. Figure 3 shows the treatment scheme for DC from Balb / c mice. Figure 4 shows the first and second incubation of T cells with alloglogic cells.

Figure 5 shows CD86 expression of the Balb / c-DC before starting the first incubation. Figure 6 shows the cytotoxicity of the CD8 + T cells, which were incubated in the first incubation with differently treated DC.

The problem is solved by cells of the recipient which are antigen-presenting cells or are cells from which antigen-presenting cells can be generated. Of particular interest in the case of the antigen-presenting cells are the dendritic cells, but also B cells. The antigen-presenting cells are to be incubated with allologic bone marrow beam planets. These grafts contain lymphocytes. In a preferred embodiment, the antigen-presenting cells can can also be incubated alone with the lymphocytes or T cells (T lymphocytes). In a preferred embodiment, the antigen-presenting cells can also be incubated alone with the lymphocytes or T cells (T-lymphocytes). Among the lymphocytes, the T cells are of particular interest because they are important reactions for the recipient / patient

trigger onen. One of the reactions is wanted and useful, the other is harmful and often leads to the death of the recipient. The useful response is anti-leukemic GvL, the harmful response is GvHD. The incubation of the antigen-presenting cells with the transplant and here in particular with the T cells occurring therein is said to switch off the GvHD reaction and to maintain only the GvL reaction. For this purpose, the antigen-presenting cells are selected or treated in such a way that they can no longer activate T cells, but can still present the recipient's alloantigens. If an alloreactive T cell encounters such an antigen-presenting cell, it is permanently shut down, or sent to programmed cell death (apoptosis), or converted into a regulatory T cell. A regulatory T cell is understood to be a T cell that prevents immune responses to specific antigens, here alloantigens. The following are ways to solve the problem:

A.

The problem is solved by the cell according to the invention, which originates from the recipient of the transplant and is characterized in that one of the functions of co-stimulatory receptors, such as a CD83 receptor and / or B7 and / or CD40 receptor, is suppressed and / or CTLA4-binding molecules are added which bind CTLA4 to T cells of the transplant and / or Fas binding molecules are added which bind the Fas to T cells of the transplant and / or TRAIL receptor binding molecules are added, bind the TRAIL receptors to T cells of the graft, or interleukin 10 (IL-10) is added. The molecules that bind CD83, CD80, CD86, CD40 block costimulatory receptors APCs. By blocking the costimulation, T cells are switched off and / or they are driven into programmed cell death (apoptosis) and / or they differentiate into regulatory T cells if the T cells simultaneously present an antigen with their T cell receptor detect. Molecules that bind Fas and / or TRAIL receptors to T cells in the transplant can cause T cells to enter programmed cell death (apoptosis)

be driven if they simultaneously recognize a presented antigen with their T cell receptor. IL-10 hinders the activation of T cells (Corinti et al 2001). The antigens recognized by this method are predominantly transplantation antigens (main histocompatibility complexes (HLA, MHC I, MHC II), rhesus factor, secondary histocompatibility antigens (minor histocompatibility antigens)), since these differ between donor and recipient of the transplant , In a preferred embodiment, the binding molecules are crosslinked, which e.g. by other binding molecules, e.g. Antibodies can happen, which in turn bind the binding molecules against CD83, CD80, CD86, CD40, CTLA4, Fas, TRAIL receptors. Networking promotes the desired effect.

B.

The cells according to the invention can also be represented in whole or in part by immature dendritic cells (DC; e.g. those derived in vitro). Due to the lack of costimulation in immature DCs, T cells (as in A.) are removed

switches and / or they are driven into programmed cell death (apoptosis) and / or they differentiate into regulatory T cells. The immature DCs behave like DCs whose costimulation (see A.) is blocked (Dhodapkar et al 2001).

C.

The cell according to the invention can also have at least one gene for a

Transfected binding molecule. Through the expression of FasL on the plasma membrane of dendritic cells, T cells (as in A.) can be forced to programmed cell death (apoptosis).

D.

The cell according to the invention can also contain at least one gene for a tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor.

Transfected binding molecule. The expression of TRAIL receptor binding molecules on the plasma membrane of dendritic cells can force T cells (as in A.) to programmed cell death (apoptosis).

E. The cell according to the invention can also be added by adding aspirin, la, 25 dihydroxyvitamin D3, la, 25 (OH) 2-16-ene-23-yne-26,27-hexafluoro-19-nor-vitamin D3 (D3 analog), vitamin D3, glucocorticoids are treated (Adorini et al 2001; Griffin et al 2001; Griffin et al 2000; Hackstein et al 2001; Penna & Adorini 2000; Penna & Adorini 2001) (WO 01/24818). As a result of this intervention, dendritic cells remain in a state which limits the immunostimulatory properties of the dendritic cells. The reduced costimulation switches off T cells (as in A.) and / or drives them to programmed cell death (apoptosis) and / or differentiates them into regulatory T cells.

F.

The cell according to the invention can also be transfected with at least one gene for a CTLA4 binding molecule. By expressing CTLA4 binding molecules on the plasma membrane of dendritic cells, T cells can be switched off and / or driven into programmed cell death (apoptosis) and / or differentiated into regulatory T cells. G.

The cell according to the invention can also be transfected with at least one gene for a CD83 binding molecule and / or for B7 (CD80 / CD86) binding molecules and / or for a CD40 binding molecule. By expressing CD83 binding molecules and / or B7 (CD80 / CD86) binding molecules and / or CD40 binding molecules in the endoplasmic reticulum and / or Golgi and / or extracellularly and / or on the plasma membrane of dendritic cells, T cells can be switched off and on / or driven into programmed cell death (apoptosis) and / or differentiated into regulatory T cells.

H.

The cell according to the invention can also be transfected with at least one gene for IL-10. Expression of IL-10 from dendritic cells prevents the dendritic cells from activating T cells (Corinti et al 2001), so that T cells can be switched off and / or driven into programmed cell death (apoptosis) and / or be differentiated into regulatory T cells.

I.

The cell according to the invention can be induced by incubation with the interferons (IFNa or IFNγ or IFNß) to synthesize tumor necrosis factor-related apoptosis inducing ligand (TRAIL). The expression of TRAIL- on the plasma membrane of dendritic cells can force T cells (as in A.) to programmed cell death (apoptosis) (Liu et al 2001; Fanger et al 1999).

One of the procedures (A.-I.) and / or any sequence of these procedures (A.-I.) is used to treat the cell according to the invention in vitro with allogeneic stem cells (eg including lymphocytes) or with allogeneic T cells or incubate with allogeneic lymphocytes from a foreign stem cell transplant donor (e.g. bone marrow). The cells mentioned, which come from the recipient of the transplant and in cancer patients, esp especially those with leukaemias that can contain cancer cells can be treated in a preferred embodiment of the method according to the invention before incubation with the transplant in such a way that these cells will not proliferate. This is preferably achieved by irradiating these cells.

(Apoptosis) driven and / or differentiated into regulatory T cells.

H.

The cell according to the invention can also be transfected with at least one gene for IL-10. Expression of IL-10 from dendritic cells prevents the dendritic cells from activating T cells (Corinti et al 2001), so that T cells can be switched off and / or driven into programmed cell death (apoptosis) and / or to be differentiated into regulatory T cells.

I. The cell according to the invention can be induced by incubation with the interferons (IFNa or IFNγ or IFNß) to synthesize tumor necrosis factor-related apoptosis inducing ligand (TRAIL). Expression of TRAIL- on the plasma membrane of dendritic cells can force T cells (as in A.) to programmed cell death (apoptosis) (Liu et al 2001; Fanger et al 1999).

One of the procedures (A.-I.) and / or any sequence of these procedures (A.-I.) is used to treat the cell according to the invention in vitro with allogeneic stem cells (eg including lymphocytes) or with allogeneic T cells or incubate with allogeneic lymphocytes from a foreign stem cell transplant donor (e.g. bone marrow). The cells mentioned, which originate from the recipient of the transplant and can contain cancer cells in cancer patients, in particular those with leukemia, can be found in ei ner preferred embodiment of the method according to the invention are treated before incubation with the graft so that proliferation of these cells will not occur. This is preferably achieved by irradiating these cells.

The non-alloreactive, but allologic T-cells positively selected by one of these procedures and / or any sequence of these procedures are presented to the patient together with the allologic stem cells (e.g. bone marrow)

infused, preferably into the bone marrow. In this way, a graft-versus-leukemia effect (GvL) is obtained, but none, or a significantly reduced graft-versus-host disease (GvHD).

The CD83 binding molecules can preferably be antibodies, monoclonal antibodies, F (ab) ₂ , single chain antibodies (scFv), and / or Fab fragments.

The proteins which have affine structures for B7 receptors can preferably be CTLA4, CTLA4 derivatives (eg CTLA4Ig), CD28, antibodies, F (ab) ₂ , scFv and / or Fab fragments.

The proteins which have structures affinity to CD40 receptors can preferably be CD40L, CD40L derivatives, antibodies, F (ab) ₂ , scFv and / or Fab fragments.

The proteins which have affine structures for CTLA4 (CD152) receptors can preferably be antibodies, F (ab) ₂ , scFv and / or Fab fragments. The proteins which have affine structures for Fas ligands can preferably be FasL, FasL derivatives, antibodies, F (ab) ₂ , scFv and / or Fab fragments. The proteins which have affine structures for TRAIL receptors can preferably be TRAIL, TRAIL derivatives, antibodies, F (ab) ₂ , scFv and / or Fab fragments.

The method according to the invention can be based on genetic engineering interventions on the patient's own cells (pre-cells: stem cells, monocytes, DCs, CD34 + stem cells, CD34 + progenitor stem cells and / or B cells). The interventions take place by means of suitable probes and bring about the reduction of CD83 molecules and possibly B7 molecules and / or CD40 molecules by hindering or preventing the formation of the molecule on the surface of the antigen-presenting cells and / or CTLA4 binding Molecules, preferably antibodies, and / or the production of Fas binding molecules, preferably FasL and / or the production of TRAIL receptor binding molecules, preferably TRAIL and / or the production of IL-10.

Transfection of the pre-cells (stem cells, monocytes, DCs, CD34 + stem cells, CD34 + progenitor stem cells and / or B cells) or antigen-presenting cells causes expression of Fas binding molecules and / or TRAIL receptor binding molecules and / or IL-10 and / or B7 binding molecules and / or CTLA4 binding molecules and / or CD83 binding molecules and / or CD40 binding molecules.

This is achieved by nucleic acids coding for Fas binding molecules and / or TRAIL receptor binding molecules and / or IL-10 and / or B7 binding molecules and / or CTLA4 binding molecules and / or CD83 binding molecules and / or CD40 binding molecules, reached.

The nucleic acids can be DNA, RNA, oligonucleotides, polynucleotides, ribozymes or peptide nucleic acids (PNA). In a preferred embodiment, the DNA contains regulatory elements such as enhancers, promoters, polyA-coding 3'-ends for the transcription of the DNA into RNA. The RNA in turn should contain regulatory elements for translating the RNA into protein.

For some of the expression products described (Fas, TRAIL receptor and CTLA4 binding molecules), a localization in the plasma membrane of the pre-cells and / or APCs is sought. For this purpose, the nucleic acids are provided with a signal sequence which effects the transport of the expression products to the plasma membrane.

In the case of other expression products described (B7, CD83 and CD40 binding molecules), in a preferred embodiment, localization in the endoplasmic reticulum, the Golgi apparatus, the Trans-Golgi network or intracellular vesicles is sought. For this purpose, the nucleic acids encode a signal sequence which causes the expression products to remain in the respective cell compartments.

The cells mentioned can be transfected ex vivo by treatment with viruses, viral vectors, bacterial vectors, plasmids which are suitable by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules into eukaryotic cells.

Said cell can be treated with viruses, viral vectors, bacterial vectors, plasmids by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules into a cell with increased production of Fas binding molecule and / or TRAIL receptor binding molecule and / or CD83 binding molecules, CD40 binding molecules, B7 (CD80 / CD86) binding molecules, CTLA4 binding molecule, IL-10 are transfected, whereby T cells which have allologic antigens, for example on MHC -Molecules are presented to the antigen- bind presenting cell, be switched off. Molecules such as antibodies, proteins, peptides, peptidomimetics, CTLA4, CTLA4 derivatives such as CTLA4Ig, CD28, CD40L and / or constituents and / or combinations of these molecules, which bind, for example, B7-1, B7-2, CD83, CD40, can be used as binding molecules , serve. These molecules hinder stimulation and / or co-stimulation of T cells which take place in the presence of an alloantigen presentation and are brought into contact with the cell according to the invention.

The molecules (Fas binding molecule, TRAIL receptor binding molecule, CD83 binding molecules, CD40 binding molecules, B7 (CD80 / CD86) binding molecules, CTLA4 binding molecule, IL-10 gene) can be generated by vehicles such as liposomes, hydrogels,

Cyclodextrins, nanocapsules, nanoparticles, bio-adhesive microspheres and / or by electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules into the cell according to the invention.

Nucleic acids can be transferred in particular by viruses, viral vectors, bacterial vectors, plasmids, which are transferred into the monoantigenic antigen-presenting cell and / or the antigen-presenting cell by electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules.

The APC or pre-cell which consists of the group consisting of stem cells, monocytes, DCs, CD34 + stem cells, CD34 + progenitor stem cells and / or B-

Cells are selected, can be incubated with an inhibitor of hypusin biosynthesis, such as GC7 (N ¹ -guanyl-l, 7-diaminoheptane), to hinder the production of functional eIF-5A, which in turn hampers CD83 biosynthesis. A medicament containing at least one cell according to the invention is also claimed according to the invention. The medicament according to the invention is preferably formulated as an infusion solution for intravenous or intraperitoneal application. The formulation is chosen so that when the drug is administered there is no significant impairment of the effectiveness of the antigen-presenting cell according to the invention.

For example, physiological saline is preferred as the infusion solution. In principle, other solutions with a pH of 5.5 to 8.5 are also suitable. Serum, for example human serum, autologous serum or serum of other species, solutions with plasma substitutes, such as polyvinylpyrrolidone, are also suitable. Typically 0.5 ml to 500 ml should be applied. These amounts and pH values are self

understandable not absolutely, but can be varied by the specialist, depending on the conditions and requirements, and adapted to the specific needs of a patient.

The antigen-presenting cell according to the invention can be used in particular for the production of a medicament for the treatment of immune reactions against allologic tissue features or for the prevention of GvHD. According to the invention, he is claimed to have a use in which the immune reactions to be treated are connected with allologic tissue characteristics, their gene sequences and / or partial sequencesή, in particular major histocompatibility complexes, MHC I, MHC II, rhesus factor, minor histocompatibility antigens (minor histocompatibility antigens) , A use is claimed according to the invention in which the binding molecules are crosslinked in a preferred embodiment, which is achieved by other binding molecules, e.g. Antibodies, which in turn bind the molecules against CD83, CD80,

Bind CD86, CD40, CTLA4, Fas, TRAIL receptors. Such cross-linking enhances the effect of these binding molecules. According to the invention, it also becomes a Use claimed in which the ex vivo selected cells are infused into the patient and the patient is then subjected to an in vivo treatment with IL-2. The cells selected ex vivo can also be incubated ex vivo with IL-2 or other stimulants such as phytohemaglutinin (PHA) and then infused into the patient. It has been shown that the GvL effect can be enhanced in this way. This effect is described in the literature (Klingemann HG 1995; Sykes M 1994; Uharek L 1998; Vourka-Karussis U 1995; Weiss L 1995; Weiss L 1999; Xun CQ 1995). The invention is illustrated by the following examples.

Examples:

objective:

Changes in the response of helper CD4 + T cells and cytotoxic CD8 + T cells (C3H mouse) against target cells such as A20 cells (lymphoma cells from Balb / c) and Balb / c-DC after they were incubated with differently treated DC. In the following, the CD8 + T cells are referred to as effector cells. A20 cells and Balb / c-DC are the target cells. The cytotoxicity assay consists of two incubations. The first is used to incubate allologic T cells (C3H) with "autologous" DC (Balb / c). In the second incubation (the actual cytotoxicity assay), the T cells are removed from the first incubation and incubated with target cells for 4 h. The target cells are lysed by allologic, cytotoxic CD8 + T cells, to an extent that depends on how many allologic T cells react against the foreign target cells. Mature and immature DC were used as DC. These should act differently on the T cells. Mature DC stimulate T cells, while immature DC turn off T cells. Mature DCs blocked in Kostimulus B7 turn off T cells. The experimental approach is shown in Fig. 1 and Fig. 2. Execution:

First incubation

BALB / c DC:

Immature DC (imDC from immature DC) were obtained from bone marrow from Balb / c-

Mice won. On day 8, half of the immature Balb / c-DC was overlaid with 10 ng TNFα per ml complete medium. (22 h) mature. Half of the mature and immature DC were irradiated (13 sec in the Irradiator OB

29). Half of all batches were then blocked with CTLA4-Ig (50 μg / ml).

This resulted in eight different approaches, which are shown in Tab. 1 and

Fig. 3 can be found again.

Tab. 1: Treatment of immature DC

C3H T cells:

The spleens were prepared from two C3H mice and the T cells isolated using MACS.

The isolated T cells were sown with a cell density of 5xl0 ⁵ / 500μl complete medium in a well of a 24-well plate and thus above sea level. incubated at 37 ° C in the incubator.

The complete medium had the following composition:

- 10 x RPMI (biochrom, F 1225)

L-glutamine 200 mM (biochrom, K 0283)

- NaHCO ₃ 7.5% (biochrom, L 1713)

- sodium pyruvate 100mM (biochrom, L 0473) - sterile water

- 10% fetal calf serum (Sigma, Lot No. 38H8408)

Mercaptoethanol (50 mM in PBS)

The complete medium was set up as follows:

100 ml of 10 x RPMI were diluted with 870 ml of water. For this purpose, 30 ml of NaHCO ₃ , 20 ml of L-glutamine and 10 ml of sodium pyruvate were pipetted. The finished medium was mixed by swirling and could then be portioned. Then the appropriate amount of fetal calf serum and 1 ml of 50 mM mercaptoethanol are added.

The DCs were prepared the day before C3H T cells with a cell density of lxl0 ^4/500 .mu.l of complete medium (per well of a 24-well plate) was added.

The DC were incubated with T cells in a ratio of 1:50 (in cell numbers 1x10 ⁴ : 5x10 ⁵ per well of a 24-well plate) for 6 days (FIG. 4).

Second incubation

After the first incubation, the cells (all cells -> also the Balb / c-DC) were removed from the well and incubated for 4 hours with the target cells A20 cells or Balb / c-DC (Fig. 4). 5xl0 ³ target cells and lxlO ⁵ effector cells were used for each approach. The total volume was 200 μl complete medium.

Preparation of the target cells

A20 cells

A20 cells were from a continuous cell culture. They are a Balb / c B cell lymphoma line. BALB / c DC

Balb / c-DC were differentiated from bone marrow cells and used as target cells in the immature stage on day 7.

All target cells were stained with CFSE (0.5 μM).

To quantify the lysed target cells, TruCount beads (Becton

Dickinson) used.

To identify the lysed target cells, the batches were stained with propidium iodide (PI) shortly before the measurement. The cells that were CFSE / PI double positive were considered for the evaluation. The target cells that died without the influence of T cells were determined as a control.

Discussion and conclusion:

The effectiveness of the B7 blockade was determined using anti-CD86 (B7-2, aCD86) monoclonal antibodies. If the CD86 signal becomes smaller in the FACS, there are fewer unblocked B7-2 molecules on the cell surface of the DC. Fig. 5 shows that the TNFα treated and therefore mature DC have a higher CD86 expression than the immature DC. CD86 could be blocked in mature and immature DC. Irradiation had no significant effect on CD86 expression.

Fig. 6 shows the percentage of the lysed target cells as a function of the irradiated / unirradiated or blocked / unblocked DC (during the first incubation), which were incubated with the effector cells.

Against the Balb / c-DC, a significantly lower cytotoxic activity can be seen in the effector cells, which were incubated during the initial incubation on DCLA4-Ig blocked DC or on immature DC (blocked and unblocked), and thus may have become anergic or tolerogenic. The highest rate of dead target cells is noted for the approaches that had A20 cells as target cells. A guess would be that this response is specific to tumor antigen.

It can be seen from the results that both immature DC (imDC) (blocked and unblocked) and blocked mature DC (DC) are able to prevent the activation of T cells on alloantigens. Afterwards, these T cells are no longer able or only able to react to these alloantigens to a greatly reduced extent.

Unmodified DC, on the other hand, have promoted the activation of alloantigen-specific T cells.

literature

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Corinti, S., Albanesi, C, la Sala, A., Pastore, S., Girolomoni, G. 2001.

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Dhodapkar, MV, Steinman, RM, Krasovsky, J., Münz, C, Bhardwaj, N. 2001. Antigen-specific inhibition of effector T cell function in humans after injection of immature dendritic cells. J Exp Med 193: 233-8. Fanger, NA, Maliszewski, CR, Schooley, K., Griffith, TS 1999. Human dendritic cells mediate cellular apoptosis via tumor necrosis factor-related a-poptosis-inducing ligand (TRAIL). J Exp Med 190: 1155-64. Griffin, MD, Lutz, W., Phan, VA, Bachman, LA, McKean, D. _., Kumar, R. 2001. Dendritic cell modulation by lalpha, 25 dihydroxyvitamin D3 and is analogous: a vitamin D receptor- dependent pathway that promotes a persistent State of immaturity in vitro and in vivo. Proc Natl Acad Sei USA 98: 6800-5. Griffin, MD, Lutz, WH, Phan, VA, Bachman, LA, McKean, DJ, Kumar, R. 2000. Potent inhibition of dendritic cell differentiation and maturation by vitamin D analogs. Biochem Biophys Res Commun 270: 701-8. Hackstein, H., Morelli, AE, Larregina, AT, Ganster, RW, Papworth, GD, et al. 2001. Aspirin inhibits in vitro maturation and in vivo immunostimulatory function of murine myeloid dendritic cells. J Immunol 166: 7053-62. Klingemann HG, PG 1995. Is there a place for immunotherapy with interleukin-2 to prevent relapse after autologous stem cell transplantation for acute leukemia? Leuk Lymphoma 16: 397-405 Kruse, M., Rosorius, O., Kratzer, F., Bevec, D., Kuhnt, C, et al. 2000a. Inhibition of CD83 cell surface expression during dendritic cell maturation by interference with nuclear export of CD83 mRNA. J Exp Med 191: 1581-90. Kruse, M., Rosorius, O., Kratzer, F., Stelz, G., Kuhnt, C, et al. 2000b. Mature dendritic cells infected with herpes Simplex virus type 1 exhibit inhibited T-cell stimulatory capacity. J Virol 74: 7127-36. Liu, S., Yu, Y., Zhang, M., Wang, W., Cao, X. 2001. The involvement of tnf-alpha-related apoptosis-inducing ligand in the enhanced cytotoxicity of ifn-beta-stimulated human dendritic cells to tumor cells. J Immunol 166: 5407-15.

Matsue, H., Matsue, K., Walters, M., Okumura, K., Yagita, H., Takashima, A. 1999. Induction of antigen-specific immunosuppression by CD95L cDNA-transfected 'killer' dendritic cells. Nat Med 5: 930-7. Min, W.P., Gorczynski, R., Huang, X.Y., Kushida, M., Kim, P., et al. 2000. Dendritic cells genetically engineered to express Fas ligand induce donor-specific hyporesponsiveness and prolong allograft survival. J Immunol 164: 161-7. Miura, Y., Misawa, N., Maeda, N., Inagaki, Y., Tanaka, Y., et al. 2001. Critical contribution of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to apoptosis of human CD4 + T cells in HIV-1 infected hu-PBL-NOD-SCID mice. J Exp Med 193: 651-60.

Penna, G., Adorini, L. 2000.1 Alpha, 25-dihydroxyvitamin D3 inhibits differentiation, maturation, activation, and survival of dendritic cells leading to impaired alloreactive T cell activation. J Immunol 164: 2405-11. Penna, G., Adorini, L. 2001. Inhibition of costimulatory pathways for T-cell 25 activation by 1,25-dihydroxyvitamin D (3). Transplant Pro 33: 2083-4.

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Claims

claims

1. antigen presenting cell available from pre-cells selected from the group consisting of stem cells, monocytes, DCs, CD34 + stem cells,

CD34 + progenitor stem cells, B-cells of a patient to be treated with an allogeneic bone marrow transplant, which may also contain blood lymphocytes, and / or from pre-existing antigen-presenting cells (APC), the pre-cells or the pre-existing ones (APC) of a treatment with binding molecules against B7 (1 and / or 2), CD83, CD40, CTLA4, Fas TRAIL receptor and / or combinations thereof on T cells, and these pre-cells or APC are incubated in vitro with allogeneic lymphocytes from a foreign stem cell transplant donor are used, and / or completely or partially immature dendritic cells (DC; e.g. in vitro derived) and these are incubated in vitro with allogeneic lymphocytes from a foreign stem cell transplant donor, - and / or the cells mentioned with at least one gene for one

Fas binding molecule is transfected and these are incubated in vitro with allogeneic lymphocytes from a foreign stem cell transplant donor, and / or the cells mentioned contain at least one gene for a tumor necrosis factor-related apoptosis-inducing ligand

(TRAIL) receptor binding molecule and these are incubated in vitro with allogeneic lymphocytes including blood lymphocytes from a foreign stem cell transplant donor, and / or the cells mentioned by adding aspirin, la, 25 dihydroxyvitamin D3, la, 25 (OH) 2-16-ene-23-yne-26,27-he- xafluoro-19-nor-vitamin D3 (D3 analogue), vitamin D3, glucocorticoids or combinations thereof dendritic cells in one Condition that limits the immunostimulatory properties of the dendritic cells and these are incubated in vitro with allogeneic lymphocytes from a foreign stem cell transplant donor,

and / or the said cells are transfected with at least one genetic sequence for interleukin 10 (IL-10) and these are incubated in vitro with allogeneic lymphocytes from a foreign stem cell transplant donor, and / or the said cells by adding interleukin 10 at the activation are prevented by T cells and these are incubated in vitro with allogeneic lymphocytes from a foreign stem cell transplant donor, and / or the named cells are added by adding interferon a or g or b (IFNa or IFNg or IFNb) to synthesize tumor cells. Necrosis factor-related apoptosis-inducing ligand (TRAIL) is initiated and these are incubated in vitro with allogeneic lymphocytes from a foreign stem cell transplant donor, and / or the said cells are transfected with at least one gene for CTLA4 binding molecules and these are transfected with allogeneic cells in vitro

Lymphocytes from a foreign stem cell transplant donor are incubated, and / or the cells mentioned are transfected with at least one gene for CD83 binding molecules and optionally for CD40 binding molecules and optionally for B7 (CD80 / CD86) binding molecules, and these are transfected in vitro with allogeneic lymphocytes from a third party Stem cell transplant donors are incubated.

2. Cell according to claim 1, wherein the cell originate from the recipient of the transplant and can contain cancer cells in cancer patients, in particular those with leukemia, before incubation with the Transplant are treated so that there is no proliferation of these cells, in particular by irradiation of these cells.

3. Cell according to one of claims 1 and / or 2, characterized in that a transfection of the pre-cells (stem cells, monocytes, DCs,

CD34 + stem cells, CD34 + progenitor stem cells and / or B cells) or APC an expression of Fas binding molecules and / or TRAIL receptor binding molecules and / or IL-10 and / or B7 binding molecules and / or CTLA4 binding molecules and / or CD83 binding molecules and / or CD40 binding molecules.

4. Cell according to at least one of claims 1 to 3 containing nucleic acids which encode Fas binding molecules, TRAIL receptor binding molecules, IL-10, B7 binding molecules, CTLA4 binding molecules, CD83 binding molecules, CD40 binding molecules or combinations thereof.

5. Cell according to at least one of claims 1 to 4, in which the CD83-binding molecules are preferably antibodies, monoclonal antibodies, F (ab) ₂ , single chain antibodies (scFv), Fab fragments or combinations thereof.

6. Cell according to at least one of claims 1 to 5, wherein the proteins which have affine structures for B7 receptors CTLA4, CTLA4 derivatives (eg CTLA4Ig), CD28, antibodies, F (ab) ₂ , scFv, Fab fragments or Combinations of these are.

7. The cell according to the invention according to at least one of claims 1 to 6, wherein the proteins which have structures affine to CD40 receptors are CD40L, CD40L derivatives, antibodies, F (ab) ₂ , scFv, Fab fragments or combinations thereof.

8. The cell according to the invention according to at least one of claims 1 to 7, wherein the proteins which have affine structures for CTLA4 (CD152) receptors are antibodies, F (ab) ₂ , scFv, Fab fragments or combinations thereof.

9. Cell according to at least one of claims 1 to 8, wherein the proteins which have structures affine to Fas receptors are FasL, FasL derivatives, antibodies, F (ab) ₂ , scFv, Fab fragments or combinations thereof.

10. Cell according to at least one of claims 1 to 9, wherein the proteins that have affine structures for the TRAIL receptor are TRAIL, TRAIL derivatives, antibodies, F (ab) ₂ , scFv, Fab fragments or combinations thereof.

11. Cell according to at least one of claims 1 to 10, characterized in that the nucleic acids are DNA, RNA, oligonucleotides, polynucleotides, ribozymes, peptide nucleic acids (PNA) or combinations thereof.

12. Cell according to at least one of claims 1 to 11, characterized in that the DNA regulatory elements such as enhancers, promoters, polyA-coding 3 'ends for transcription of the DNA into RNA contains the RNA regulatory elements for translating the RNA into protein.

13. Cell according to at least one of claims 1 to 12, wherein the nucleic acids code for a signal sequence or the expression products have a signal sequence which brings about the transport of the expression products to the plasma membrane.

14. Cell according to at least one of claims 1 to 13, wherein the nucleic acids code for a signal sequence or the expression products ne have a signal sequence which causes the expression products to remain in the endoplasmic reticulum, the Golgi apparatus, the Trans-Golgi network or intracellular vesicles.

15. A method for producing a cell according to one of claims 1 to 14, from stem cells, monocytes, DCs, CD34 + stem cells, CD34 + progenitor stem cells, B cells or combinations thereof of a patient to be treated with a bone marrow transplant to generate antigen-presenting cells which are subject to treatment with binding molecules against B7, CD83, CD40, CTLA4, Fas, the TRAIL receptor on T cells or combinations thereof, and the DC in vitro with allogeneic stem cells or with allogeneic T cells or with allogeneic lymphocytes of a foreign person Incubate stem cell graft donors to turn off alloreactive T cells, which interact intensively with the dendritic cells via complex adhesion systems and TCR / MHC complexes, by blocking the costimulation by the dendritic cells and / or turning them into programmed cell death ( To force apoptosis) and / or to differentiate them into regulatory T cells, - and / or quite o to use the partially immature dendritic cells and incubate them in vitro with allogeneic stem cells or with allogeneic T cells or with allogeneic lymphocytes from a foreign stem cell transplant donor in order to intensify the alloreactive T cells> via complex adhesion systems and TCR / MHC complexes Interact with the dendritic cells, which are missing in immature DCs

Switch off costimulation and / or force them into programmed cell death (apoptosis) and / or differentiate them into regulatory T cells, and / or transfect the cells mentioned - with at least one gene for a Fas binding molecule and use them in vitro incubate allogeneic stem cells or with allogeneic T cells or with allogeneic lymphocytes from a foreign stem cell transplant donor in order to increase the alloreactive T cells, which enter into intensive interaction with the dendritic cells via complex adhesion systems and TCR / MHC complexes, to be forced into programmed cell death (apoptosis) by the FasL expressed by the dendritic cells on the plasma membrane, and / or the cells mentioned with at least one gene for the (eg human) tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor binding molecule and transfect this in vitro with allogeneic stem cells or with allogeneic T cells or with allogeneic lymphocytes from a foreign stem cell -Graft donors to incubate the alloreactive T cells that have complex adhesion systems and

TCR / MHC complexes enter into intensive interaction with the dendritic cells, by forcing the TRAIL receptor binding molecule expressed by the dendritic cells on the plasma membrane into programmed cell death (apoptosis), and / or by adding aspirin, la, 25 dihydroxyvitamin D3, la, 25 (OH) 2-16-ene-23-yne-26,27-hexafluoro-19-nor-vitamin D3, vitamin D3, glucocorticoids to keep dendritic cells in a state that limits the immunostimulatory properties of the dendritic cells and incubate them in vitro with allogeneic stem cells or with allogeneic T cells or with allogeneic lymphocytes from a foreign stem cell transplant donor in order to incubate the alloreactive T cells via complex adhesion systems and TCR / MHC Complex intensive interaction with the dendritic cells, switch off by the reduced costimulation and / or force them into programmed cell death (apoptosis) and / or regulato them differentiate T cells, and / or transfect the cells with at least one gene for interleukin 10 (IL-10), which limits the immunostimulatory properties of the dendritic cells and these in vitro with allogeneic stem cells or with allogeneic T cells or with Incubate allogeneic lymphocytes from a foreign stem cell graft donor to the alloreactive T cells, which have complex adhesion systems and TCR / MHC Enter into complex intensive interaction with the dendritic cells, switch off, force them into programmed cell death (apoptosis), differentiate them into regulatory T cells or a combination of this measure and / or the said cells are mixed with interleukin 10, which has the immunostimulatory properties of the dendritic cells Cells and incubate them in vitro with allogeneic stem cells or with allogeneic T cells or with allogeneic lymphocytes from a foreign stem cell transplant donor in order to intensively interact with the alloreactive T cells, which have complex adhesion systems and TCR / MHC complexes dendritic cells enter, switch off, force them into programmed cell death (apoptosis), differentiate them into regulatory T cells or combinations of these measures, and / or transfect the cells mentioned with at least one gene for CTLA4 binding molecules and these in vitro with allogeneic stem cells or incubate with allogeneic T cells or with allogeneic lymphocytes from a foreign stem cell transplant donor in order to allow the alloreactive T cells, which interact intensively with the dendritic cells via complex adhesion systems and TCR / MHC complexes, by means of the dendritic cells to switch off the CTLA4 binding molecule expressed on the plasma membrane, to force it into programmed cell death (apoptosis), to differentiate it into regulatory T cells or combinations of these measures and / or the named cells with at least one gene for CD83 binding molecules, for CD40 Transfect binding molecules for B7 (CD80 / CD86) binding molecules or combinations thereof and incubate them in vitro with allogeneic stem cells, with allogeneic T cells or with allogeneic lymphocytes from a foreign stem cell transplant donor in order to generate the alloreactive T cells via complex adhesion systems and TCR / MHC complexes intensive interaction with the dendritic n cells enter, through which extracellularly expressed by the dendritic cells in the endoplasmic reticulum, Golgi, on the plasma membrane CD83 and / or CD40 binding molecules, B7 binding molecule, which binds the CD83 and / or CD40 and / or B7 of the dendritic cells, to switch them off, to force them into programmed cell death (apoptosis) or to differentiate them into regulatory T cells or Combinations thereof, which are positively selected by one of these procedures and / or any sequence of these procedures, non-alloreactive, allologic T cells are infused with the allologic stem cells, preferably into the bone marrow, for a graft-versus-leukemia effect (GvL). to obtain.

16. The method according to claim 15, wherein said cells ex vivo by treatment with viruses, viral vectors, bacterial vectors, plasmids by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules into eukaryotic Cells is transfected.

17. The method of claim 15 and / or 16, wherein said cell by treatment with viruses, viral vectors, bacterial vectors, plasmids by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods and / or other techniques for the introduction of molecules in a cell with increased production of Fas binding molecule, RAIL receptor binding molecule, CD83 binding molecules, CD40 binding molecules, B7 (CD80 / CD86) binding molecules, CTLA4 binding molecule, IL-10 or combinations thereof, whereby T - Cells, which have allolog antigens, for example are presented on MHC molecules, to which the antigen presenting cell binds, are switched off.

18. The method according to at least one of claims 15 to 17, wherein molecules such as antibodies, proteins, peptides, peptidomimetics, CTLA4, CTLA4 derivatives such as CTLA4Ig, CD28, CD40L, components or combinations of these molecules, for example B7-1, B7-2 , CD83, CD40 which stimulate in the presence of an alloantigen presentation and / or co-stimulation of T cells that are brought into contact with the cell according to the invention.

19. The method according to at least one of claims 15 to 16, wherein the molecules (Fas binding molecule, TRAIL receptor binding molecule, CD83 binding molecules, CD40 binding molecules, B7 (CD80 / CD86) binding molecules, CTLA4 binding molecule, IL-10 Gene) or combinations thereof according to claim 12 by vehicles such as liposomes, hydrogels, cyclodextrins, nanocapsules, nanoparticles, bio-adhesive microspheres, by electroporation techniques, iontophoresis, ballistic methods or combinations of these techniques for introducing molecules into the invention Cell are transferred.

20. The method according to at least one of claims 15 to 19, wherein nucleic acids by viruses, viral vectors, bacterial vectors, plasmids transferred by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods or combinations of these techniques for the introduction of molecules into said cells become.

21. Pre-cell selected from the group consisting of stem cells, monocytes, DCs, CD34 + stem cells, CD34 + progenitor stem cells, B cells or combinations thereof, characterized in that an inhibitor of hypusin biosynthesis, such as GC7 (N ¹ -guanyl- 1,7-diaminoheptane) is used to hinder the production of functional eIF-5A, which in turn hampers CD83 biosynthesis.

22. Medicament containing at least one cell according to the invention or according to at least one of claims 1 to 14.

23. Medicament according to claim 22, wherein at least one cell according to the invention is formulated for ex vivo application.

24. Use of at least one cell according to the invention according to at least one of claims 1 to 14 for the manufacture of a medicament for the treatment of immune reactions against allologic tissue features.

25. Use according to claim 24, wherein the immune reactions to be treated are associated with allologic tissue characteristics, their gene sequences and / or partial sequences, in particular major histocompatibility complexes, MHC I, MHC II, rhesus factor, minor histocompatibility antigens (minor histocompatibility antigens).

Use according to claims 1 to 25, wherein the binding molecules are crosslinked in a preferred embodiment, which is prevented by other binding molecules, e.g. Antibodies can happen, which in turn bind the binding molecules against CD83, CD80, CD86, CD40, CTLA4, Fas, TRAIL receptors.

27. Use according to claim 1 to 26, wherein the ex vivo selected cells are infused into the patient and the patient is then subjected to an in vivo treatment with IL-2.

28. Use according to claim 1 to 26, wherein the ex vivo selected cells are incubated ex vivo with IL-2 or other stimulants such as phytohemaglutinin (PHA) and then infused into the patient.