WO2003006636A1

WO2003006636A1 - Reduction of the stimulatory capacity of antigen-presenting cells

Info

Publication number: WO2003006636A1
Application number: PCT/EP2002/007740
Authority: WO
Inventors: Ahmed Sheriff
Original assignee: Genethor Gmbh
Priority date: 2001-07-12
Filing date: 2002-07-11
Publication date: 2003-01-23

Abstract

The invention relates to a method for reducing immune reactions. The inventive method is characterized by manipulating the stimulatory properties of antigen-presenting cells and optionally at the same time inducing the antigen-presenting cells to present defined antigens.

Description

Reduction of the stimulation ability of antigen presenting cells

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

For almost 10 years, gene therapy methods have been developed and used to treat various diseases and in animal models. So far, however, they have not yet been put into daily practice. Most of the time, this is the treatment of genetically determined, serious diseases for which other therapies are not available. Gene therapy is also used to treat severe and also non-treatable cancer.

Experimental medicine has so far paid little attention to the treatment of allergies and autoimmune diseases using genetic engineering methods.

General information about allergies

Allergies cause considerable healthcare costs in industrialized countries. After all, it is estimated that at least 20% of the population is allergic to any substance. Most suffer from allergic rhinitis, which includes hay fever in particular, or bronchial asthma: they sneeze or gasp for breath after inhaling certain pollen or other generally harmless substances. Many children and some adults are also allergic to food. Others experience rashes or even an allergic shock after taking medication such as penicillin. Still others, bee stings cause severe local swelling or severe systemic disorders that affect the entire organism. In extreme cases, allergic attacks can even lead to death. Immediate medical care for asthma patients in the United States swallowed up an estimated $ 3.6 billion in 1990, about one percent of all costs in the. Healthcare identified.

It is now known that some of the cellular and molecular interactions in allergic reactions are often similar, regardless of which substances the individual responds to and what symptoms he develops. Certain side effects of allergies usually only occur when the immune system is fighting parasites. The body responds to parasites as well as allergens with the massive production of molecules called immunoglobulin E antibodies (IgE). The production of these antibodies is induced by T helper cells, which in turn are activated by B cells.

sensitization

Different allergens cause different symptoms, among other things, because they come into contact with the immune system in different parts of the body. In the upper airways, the misguided immune response causes sneezing and a stuffy nose. In contrast, the bronchi in the lower airways can narrow and become mucous, so that typical asthmatic symptoms appear. Accordingly, immune activities in the tissues of the gastrointestinal tract cause nausea, abdominal cramps, diarrhea or vomiting. After all, an allergen that gets into the blood in some way can trigger anaphylaxis: an allergic reaction in regions of the body that are far from its point of entry. Severe anaphylactic shocks can upset all normal bodily functions and can be fatal.

Even if the allergic reactions appear differently, they are always set in motion by the same mechanism: sensitization. One-time contact with an allergen, typically a protein, can suffice for this. In the airways or other tissues, the allergenic substance meets so-called phagocytes or macrophages. These absorb the foreign substance, disassemble it and present the fragments on the cell surface with MHC II molecules. In the further course, some T-helper lymphocytes recognize the fragments presented and bind to it. The T helper lymphocytes are activated by the macrophages and then in turn activate some B lymphocytes, which also recognize the allergen. The B cells then mature into plasma cells that produce antibodies. First of all, these are antibodies of the so-called IgM type; from a certain point in time, however, the plasma cells switch to IgE antibodies.

It may take days or weeks for the antibodies to be made, and the allergen that started their production may already be gone. Not the IgE molecules. With their Fc region, they attach themselves to IgE receptors of two different classes of cells of the immune system. One class is mast cells, which are usually located in the body tissues near blood vessels and epithelial cells. There is contact with the outside world via the epithelium (this also includes the epithelium of the respiratory tract and gastrointestinal tract). IgE antibodies also bind to basophilic granulocytes (basophils). However, these cells circulate in the bloodstream.

Once production of the IgEs begins, it apparently lasts for months, sometimes even years. As a result, they constantly occupy IgE receptors on mast cells and basophils - ready to take immediate action the next time they come into contact with allergens.

Acute symptoms

So while the first encounter with an allergen does not cause symptoms even in people who later turn out to be allergy sufferers, the second exposure initiates a stage of the hypersensitivity reaction, which also appears externally. The allergy-causing substance binds to the IgEs of the mast cells within seconds of contact with human tissue. If it attaches to two or more IgE molecules at the same time, it forms a bridge between them. Such cross-links bring the affected IgE receptors closer together, and this activates the cell so that it releases highly effective substances that directly produce allergic symptoms. (The release can also affect others Species are caused; one only speaks of allergic reactions if IgEs are involved). The most important of these substances is histamine. It can stimulate the formation of mucus in the epithelia and thus contribute to the blockage of the air passages, as well as the smooth muscles that wrap around the bronchial tubes and intestines like an elastic band. It is also able to dilate the fine blood vessels and make them more permeable so that fluid can seep into the tissue. The result is redness and swelling. If these vascular changes affect large parts of the body, they can cause fatal circulatory failure: With such a shock, the blood pressure suddenly drops so much that the oxygen supply to the heart and brain is no longer guaranteed.

The second group of mediators consists mainly of prostaglandins and leukotrienes. They are only released after the allergen molecules have attached to the IgEs on the cells. Like histamine, they narrow the bronchi and dilate the blood vessels. However, their effects last longer.

In addition, stimulated mast cells emit a variety of potentially toxic enzymes. They also appear to release cytokines that regulate the activities of other immune cells. Treatment: 1. Allergic rhinitis

Antihistamines are usually effective and still serve as standard therapy. The latest variants can no longer easily cross the blood-brain barrier and no longer make the patient tired. If antihistamines remain ineffective in severe inflammation, inhalable corticosteroids, which are usually prescribed to relieve chronic inflammation in asthma, often help.

In severe cases, immunotherapy or hyposensitization (also known as allergy shots or desensitization), which was introduced in 1911, can provide relief in the long term. With this treatment, doctors inject patients with increasing doses of the allergen to which they are sensitive. The dose is decisive in all cases: Too little Allergen gives no tolerance. In addition, protection is rarely complete. 2. Asthma

Bronchodilators are the most widely used drugs for asthma. They relieve the symptoms caused by histamine and other bronchoconstrictors very quickly, but are unlikely to affect the underlying inflammation. In addition, their excessive use can cause a back reaction of the body, so that after their effects have subsided, the respiratory flow is more restricted than before. In addition, the methods listed under 1. are used. 3. Anaphylactic reactions

Insect bites cause anaphylaxis in some people. In severe cases these lead to death e.g. Circulatory failure or suffocation. Any severe anaphylaxis - whether it occurs for the first time or for the next time - is an emergency, in which an attempt must first be made to control the most threatening symptoms. This is usually done by injecting adrenaline, which inhibits the release of mediators, opens the airways and counteracts the dilation of the blood vessels. One can act preventively by immunizing with the poison of the health-threatening insect.

New approaches

The animal model is currently testing DNA-based immunization with an allergen (Der p 5) of the mite Dermatophagoides pteronyssinus. This immunization results in the production of IgG, but not IgE, and results in a 90% reduction in the amounts of specific IgE, which was caused by classic sensitization with Der p 5 and alum as adjuvant or allergen-induced rhinitis.

Another strategy is to use humanized anti-IgE monoclonal antibodies against the FceRI binding region for IgE. This prevents the binding of IgE to the IgE receptor, so that no mediators of the allergic reaction from mast cells or basophils can be released. This strategy has been used in clinical trials in patients with allergic rhinitis and allergic asthma have shown that these antibodies are well tolerated and reduce the allergic reactions.

(see also LM Lichtenstein: "Allergy and immune system"; in spectrum of science special: The immune system; 1994; 74-83; SK Huang, KY Chua and KH Hsieh: allergen gene transfer. Current Opinion in Immunology 1997; 800-804; C. Heusser and P. Jardieu: Therapeutic potential of anti-IgE antibodies. Current Opinion in Immunology 1997; 805-814).

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 activation of T-helper lyran phocytes. 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. In the case of other tissues, the very polymorphic main histocompatibility complexes (MHC) have to be compared with one another, since these almost always trigger the immune reaction. Unfortunately, perfect matching of the MHCs is almost impossible except for relatives.

Stem cell transplants for leukemia

Leukemia is cancer of the blood cells. 50 out of 1 million people develop leukemia each year. During the course of leukemia, the body produces large amounts of abnormal blood cells. In most types of leukemia, the abnormal cells are white blood cells. The appearance and functionality of the leukemia cells differs from that of normal blood cells. leukemias

There are several types of leukemia. Usually there are roughly two classes. One is determined by the speed with which the disease develops and worsens. The other is determined by the type of blood cell affected.

Leukemia is either acute or chronic. In acute leukemia, the abnormal blood cells are 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 increases more slowly than in acute leukemia. Chronic leukemia gradually worsens the disease.

Leukemia can appear in one of two main types of white blood cells: lymphoid cells or myeloid cells. Lymphoid cells affect lymphoid leukemia. If the myeloid cells are affected, the disease is called myeloid leukemia.

The most common leukemias are:

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

- Acute myeloid leukemia (AML) occurs in adults and children. This type of leukemia is also 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. Leukaemic cells are abnormal cells that cannot function as normal blood cells. You are unable to fight infections. Because of this, people with leukemia often develop infections associated with 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. These anemic side effects put additional strain on the patients.

Like all blood cells, leukemic cells migrate through the body. Depending on the number of abnormal cells and the location where these cells collect, patients with leukemia can have a number of symptoms.

With acute leukemia, the symptoms appear and worsen quickly. In chronic leukemia, the symptoms appear late. When symptoms appear, they are usually mild at first and gradually worsen.

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, 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 accumulate 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, and whether the leukemia has been treated before. The patient's age, symptoms and general health are also important.

Acute leukemia must be treated immediately. However, 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 receive additional radiation and / or receive a bone marrow transplant (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. Then it is removed before treatment with cytotoxic substances and treated ex vivo to treat leukemic cells remove. After that, a hospital stay of several weeks is necessary so that the graft can produce sufficient lymphocytes again. In the meantime, patients need to be protected from infection.

The biological therapies include treatments with substances that influence the immune response to cancer. Interferons, interleukins and colony stimulating factors are e.g. used in some types of leukemia. They are usually combined with chemotherapy or bone marrow transplantation.

Bone Marrow Transplantation Patients with a bone marrow transplant are at increased risk of infection, bleeding, and other side effects from the high doses of 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 the bone marrow transplants is explained below using the example of chronic myeloid leukemia:

Allologic bone marrow transplantation (BMT) has 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 creates the bcr-abl oncogene, the only factor that is absolutely necessary for the development of the disease (Daley et al 1990). Compared to other tumor types Pen 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 to 30 years, anti-tumor responses from allologic bone marrow transplants (allo-BMT) and, most recently, donor leukocyte infusions [Donor leukocyte infusion (DLI)] have been used clinically to treat 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 crude 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 an up to 70% leukemia-free survival rate for transplant recipients (Clift & Anasetti 1997). Treatment of rejection reactions

Although the immune response makes organ transplantation difficult, there are few alternatives to organ failure. The use of potent immunosuppressive drugs, especially cyclosporin A and FK-506, which prevent the activation of T cells, makes organ transplants successful. Nevertheless, here at e.g. Autoimmune diseases have some problems because the suffering that has destroyed your own organ also destroys the foreign organ. The suppression of the immune system also increases the risk of developing cancer or infections. The procedure is also very expensive (Janeway and Travers 1997).

Co-stimulatory molecules

The product of the mouse PD-1 gene (ACCESSION NM_021893), a member of the IgG superfamily, is a receptor that is common in many tissues. Analyzes carried out by flow cytometry and immunoprecipitation with the monoclonal antibody J43mAk showed that the PD-1 gene product is a 50 to 55 kDa membrane protein. The PD-1 protein appears to be highly glycosylated because the calculated molecular weight of the amino acid sequence is 29310 Da. Normal mouse lymphoid tissue such as thymus, spleen, lymph nodes and bone marrow contain only a small number of PD-1 positive cells. Nevertheless, a significant PD-1 positive population appears in thymocytes as well as on T cells in spleen and lymph nodes due to the in vivo treatment with anti-CD3 monoclonal antibodies. Furthermore, PD-1 antigen expression was strongly induced by in vitro stimulation in various subgroups of thymocytes and spleen T cells, either with anti-CD3 mAb or Concavalin A, which can cause T cells to activate as well as to die , PD-1 expression on spleen B cells was similarly stimulated with anti-IgM Ab. In contrast, PD-1 was not significantly expressed after treatments such as growth factor deprivation, dexamethasone or lipopolysaccharide. These results suggest that the expression of PD-1 is strictly regulated and by signal transduction via the antigen receptor is induced and does not exclude the possibility that the PD-1 antigen plays a role in the clonal selection of lymphocytes, although PD-1 expression is not required for the normal course of apoptosis (Agata et al 1996; Ishida et al 1992; Nishimura et al 1996). The PD-1 receptor induces the decrease in immune responses and its loss leads to the breakdown of tolerance in peripheral tissues. PD-1 deficient mice develop lupus-like proliferative arthritis and glomerulonephritis with predominant IgG3 deposition during aging. PD-1 is structurally similar to CTL-associated antigen 4 (CTLA-4), which binds B7-1 and B7-2 and plays a crucial role in the maintenance of T cell homeostasis [for reviews, see (Sperling & Bluestone 1996; Thompson & Allison 1997)]. PD-1 does not contain the MYPPPY motif, which is critical for B7-1 and B7-2 binding. The extracellular region of PD-1 and CTLA-4 each consist of an IgV domain with 23% identity to one another. A ligand (PD-Ll) exists for PD-1. The binding of PD-1 by PD-Ll leads to the inhibition of TCR-mediated T cell proliferation and cytokine secretion (Freeman et al 2000).

The human and mouse PD-Ll molecule [EMBL / GenBank / DDBS under accession numbers. AF233516 and AF233517, (Freeman et al 2000)] are members of the B7 gene family and have a similar structural organization, consisting of an IgV and an IgC domain in the extracellular region (Boussiotis et al 1996), a hydrophobic transmembrane domain, followed by a short one , charged intracellular region. Human PD-L1 is identical to B7-H1 at 290 amino acids, which is reported to activate T cell stimulation (Dong et al 1999). The mouse PD-Ll cDNA encodes a polypeptide with 70% amino acid identity to the human PD-Ll. PD-Ll has amino acid identities of 21, 20, and 23% to B7-1, B7-2, and ICOS (ligand of inducible co-stimulator) (Boussiotis et al 1996; Ling et al 2000; Swallow et al 1999; Yoshinaga et al 1999). The expression patterns of B7-1, B7-2, and PD-Ll are different. B7-2, one of the ligands of CD28 and CTLA-4, is constitutive on monocytes expressed. However, the constitutive expression of B7-1 and B7-2 cannot be observed in any organ. B7-1 and B7-2 expression can be induced in dendritic cells, macrophages and B cells (Boussiotis et al 1996), as well as some types of fibroblasts and epithelial cells. In contrast, PD-Ll is constitutively expressed by non-lymphoid, parenchymal organs such as the heart, placenta, skeletal muscles and lungs, but not the small intestine (Dong et al 1999). PD-Ll is also expressed in some cancers. These tumors may be using PD-Ll to inhibit anti-immune responses. Since PD-1 is expressed on both activated T and B cells, the expression of PD-Ll in non-lymphoid tissues as well as on dendritic cells could enable the regulation of potentially autoreactive lymphocytes. This could be an important mechanism to limit T and B cell activity in the heart, lungs, kidneys and placenta where PD-Ll is highly ex-expressed (Freeman et al 2000).

PD-1 Ligand 2 (PD-L2) is a second ligand for PD-1. Binding of PD-1 to PD-L2 dramatically inhibits T cell receptor (TCR) mediated proliferation and cytokine production by CD4 T cells. At low antigen concentration, PD-L2PD-1 interactions inhibit strong B7-CD28 signals. At high antigen concentrations, they reduce cytokine production but do not inhibit T cell proliferation. PD-LPD-1 interactions lead to cell cycle remains in Go / Gi (cell cycle phases) but do not increase the number of dead cells. PD-L2 expression on APCs is increased by treatment with interferon g and could also be detected in some other tissues and tumor cell lines. PD-Ll and PD-L2 thus have overlapping functions (Latchman et al 2001). mPD-L2 (also known as Protein AF142780) encodes a polypeptide with 38% amino acid identity to mPD-Ll. Murines and human PD-L2 have 70% amino acid identity. The five members of the B7 family, B7-1, B7-2, ICOS-L, PD-Ll and PD-L2, have 21-27% amino acid identity and a structural organization that consists of a signal sequence, an IgV-like one IgC-like and a transmembrane domain and a short one cytoplasmic "tail" exists. The cytoplasmic part of PD-Ll is conserved between mice and humans, which contrasts with the poor preservation of the cytoplasmic part of PD-L2 from humans and mice (Latchman et al 2001). The PD-L2 tissue distribution is similar to that of PD-Ll. There is expression in the placenta, heart, pancreas, lungs and liver, low expression in the spleen, lymph nodes and thymus, no expression in unstimulated monocytes, but this could be induced by interferon gamma. However, the kinetics of this induction is slower than that of PD-Ll. Functional, mature dendritic cells (DCs) arise from circulating progenitor cells after a refining 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. The functional significance of CD83 is unknown. Its increasing presence during the ripening indicates an important function (Berchtold et al 1999).

Herpes simplex virus manages to suppress the expression of CD83 in an unknown 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, the 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 involved in the export pathway of specific RNAs from the cell nucleus. eIF-5A is the only cellular protein known to contain the unusual amino acid hypusin. This modification seems for the Cell division to be necessary. Hypusin modification is a spermidine-dependent post-translational reaction that is catalyzed by two enzymes. This involves the transfer of the aminobutyl group of spermidine to the e-NH2 group of lysine at position 50 in eIF-5A by 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 has also been shown that the eIF-5A protein collects on the nucleoplasmic side of the nuclear pore complex to interact with the general nuclear export receptor CRM1 and to migrate from the nucleus into the cytoplasm in mammalian cells. Investigations of the amount of eIF-5A mRNA in human cells showed that eIF-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-1,7-diaminoheptane) j is used, the expression of CD83 and thus the full stimulatory activity of mature DCs are inhibited (Kruse et al 2000a).

General information on autoimmune diseases

Autoimmune diseases are chronic diseases. They include rheumatism in various clinical forms, diabetes, multiple sclerosis, certain forms of heart muscle inflammation and thyroid diseases. The range of autoimmune diseases can be continued, although approximately 90% epidemiologically only make up a small percentage. The clinical course of autoimmune diseases, even with one and the same diagnosis, can be very different. Relapsing courses of the disease are sometimes observed. Accordingly, the treatments are individually designed by the doctor. Some of the autoimmune diseases are antibody-mediated. In immunology, this means that the organism produces antibodies for mostly unknown reasons, which are directed against the body's own cellular structures (autoantigens). Once these antibodies have been formed, their interaction and binding to the respective organ or cell-specific structures trigger a cell and tissue-destroying reaction of the immune system. Ultimately, this leads to the clinical picture of the disease (Steinman 1994).

An example of this is the thyroid disease Graves' disease, but also a form of cardiac muscle inflammation that leads to dilated cardiomyopathy. In some cases, both the targets against which the autoantibodies are directed and the autoantibodies themselves are precisely characterized.

WO-A-00/66715-A-, EP 1 179 587 and PCT / EP 02/03292 disclose methods for reducing specific immune reactions by means of cells presenting genetically engineered antigen. These are applications with the same goals as those described here, but without taking CD83 into account.

The problem on which the invention is based is, inter alia, to provide genetic engineering, therapeutically usable products for the reduction of specific immune reactions in which targets (antigens, autoantigens) and antibodies, autoantibodies are known.

The problem is solved by the antigen presenting cell according to the invention, which can also present certain antigens beforehand (monoantigenic antigen presenting cell) and is characterized in that in the antigen presenting cell and / or monoantigenic antigen presenting cell one of the functions of co-stimulatory receptors, like a CD83 Receptor and optionally B7 and / or CD40 receptor is suppressed and / or optionally CTLA4 binding molecules are produced and / or optionally PD-1 binding molecules preferably antibodies are produced. The cell according to the invention is preferably an antigen presenting cell (APC).

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

Within the immune system, the antibodies normally play an important role as specific defense molecules ("humoral immune response"). They are produced by mature B lymphocytes. However, the induction and production of soluble antibodies is not independent of the remaining cells in the immune system; rather, this humoral immune response is controlled by other cells in the immune system. Antibody production - and thus also the production of pathological autoantibodies - cannot be properly maintained without the help and mediation of APCs and T helper lymphocytes, which in turn activate the B lymphocytes in an antigen-specific manner.

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

If the antigen is a bacterial protein, for example, the immune response is useful for the organism. However, if the antigen is an endogenous structure, one speaks of a (pathological) autoimmune 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 monocyte with the T helper lymphocytes) is a costingulatory receptor called B7. The antigen presenting cell according to the invention can have an increased expression of an antigen, preferably by transfection of an antigen presenting cell with nucleic acid-containing material, the antigen presenting cell presenting essentially only predefined antigens. The gene therapy method according to the invention is based on genetic engineering interventions on the patient's own antigen presenting 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 possibly CTLA4 binding Molecules, preferably antibodies, and / or possibly the production of PD-1 binding molecules, preferably PD-L1 and, if appropriate, a strong presentation of the autoantigen at the same time.

Reduction of specific antibody production through gene therapy

The production of pathological immune reactions is specifically ended by the manipulation of antigen presenting cells and the resulting shutdown of pathological T cells. Suppression of CD83 expression interferes with the T cell stimulatory properties of DCs, which are the most potent APCs in this regard. CD83-binding molecules can be used for this. These can trap CD83 during transport through the cell or bind to the plasma membrane and thus prevent CD83 from interacting with other molecules. Molecules such as antibodies or a CD83 ligand can be used to bind to CD83. In addition, the formation of CD83 can be prevented by hindering the formation or function of eIF-5A. Molecules such as antibodies can be used to bind to eIF-5A. An inhibitor of hypusin modification [eg GC7 (N ¹ -guanyl-l, 7-diaminoheptane)] can also be used to prevent the formation of functional eIF-5A. Antisense technology and cosuppression also lend themselves to preventing the formation of CD83 and / or eIF-5A. In addition, it can optionally be used that T cells produce the receptor PD-1. When bound to PD-Ll, this ensures that T cells no longer proliferate. Binding of PD-1 can thus shut down T cell responses. Binding molecules other than PD-Ll can also be used for this.

If necessary, additional use can be made of the fact that T cells produce two receptors that recognize B7 (1 or 2). The one receptor is e.g. B. CD28, the other is e.g. B. CTLA4. CD28 has a stimulating effect, while CTLA4 does not. The contact of CTLA4 with B7 switches off the respective T cell. This then either goes into apoptosis via programmed cell death, is switched off, or is converted into a tolerance-producing T cell. In this case, the tolerance is generated against the specific antigen recognized by this T cell. (Gribben et al., 1994; Judge et al., 1999; Lee et al., 1998; Lin et al., 1998; Metzler et al., 1997; Olsson et al., 1999; Oosterwegel et al., 1999a; Oosterwegel et al., 1999b; Peach et al., 1994; Walunas et al., 1996; Wu et al., 1997; Wulfing and Davis, 1998).

For binding to CTLA4 instead of CD28, molecules such as antibodies are used which bind to CTLA4 but not to CD28. These can then induce the T cell response promoted by CTLA4. These CTLA4 binding molecules can preferably be antibodies.

The co-receptor B7, without which the antigen presentation or the induction cascade for antibody production does not start, can optionally be additionally suppressed in order to suppress preferential binding of CD28. The co-receptor is two different co-receptors called CD80 (B7-1) and CD86 (B7-2). Their structures are known.

After the antibody production has been switched off, the autoantibodies circulating in the blood disappear due to the natural degradation and the lack of replenishment. After the in vitro manipulation of the antigen presenting cells, the cells are returned to the patient's bloodstream. The genetically modified antigen-presenting cells now switch off the pathological T-helper lymphocytes in the organism. The genetically modified cells compete directly with the APCs already present in the organism (preferably the bloodstream and lymphatic system), which normally activate the T helper lymphocytes and thus the antibody production.

Antigen presentation in genetically modified cells presenting antigen

The cDNA of a protein that causes the production of pathological autoantibodies as an autoantigen is integrated into the genome of the APC. This genetic information then serves to overproduce the autoantigen. Peptides of this autoantigen are then preferably presented on MHC II and / or MHC I. MHC II presents the peptides to the T helper cells and tries to find those that specifically recognize these presented peptides. If CD83 cannot function at the same time and may additionally be activated by a PD-1-binding molecule PD-1 instead of CD28 and possibly additionally a CTLA4-binding molecule CTLA4 is activated instead of CD28 and possibly additionally the co-receptor B7 does not function the T helper cells can be shut down and possibly suffer premature cell death or become regulatory T cells.

All genetically modified antigen presenting cells present the autoantigen on most of their MHC II complexes, whereas in vivo only very few "normal" APCs present the autoantigen and then only on a few MHC II complexes. The aim of the treatment is to displace the "normal" APCs which present the autoantigen and activate the T helper cells in vivo by the genetically modified antigen which is programmed to switch off the antibody production or T cell response. The antigen presenting cell and / or monoantigenic antigen presenting cell can in particular show an increased number of homing receptors, such as CD44. Overexpression of homing receptors will show the modified antigen-presenting cell the way into the lymph nodes, as a result of which the genetically modified antigen-presenting cells multiply and accumulate faster in lymph nodes. The lymph nodes are where most of the responses of the adaptive immune system are triggered. There, the genetically modified antigen presenting cells therefore have an effect many times higher than outside the lymph nodes.

In a preferred embodiment of the cell presenting the antigen according to the invention, a transfection brings about an increase in the number of homing receptors and or an increase in the number of PD-1-binding molecules and / or a suppression of functions of the CD83, B7, CD40 receptors and / or an increase in the number of CTLA4 binding molecules.

The formation of functional eIF-5A can be prevented with inhibitors of hypusin synthesis.

The PD-1 binding molecules can preferably be PD-Ll, PD-L2, antibodies, monoclonal antibodies. These can be such that they remain in the plasma membrane of the cell.

The CTLA4 binding molecules can preferably be antibodies, monoclonal antibodies. These can be such that they remain in the plasma membrane of the cell. In particular with antisense nucleic acids, the eIF-5A, CD83, B7 and / or CD40 receptor expression in the cell presenting the antigen according to the invention can be prevented or reduced.

Alternatively, the expression of the elF-5A, CD83, B7 and / or CD40 receptors can be suppressed by nucleic acids by co-suppression in the cell presenting the antigen according to the invention. The cell presenting the antigen according to the invention can contain or be transfected with nucleic acids which bring about an expression of proteins or peptides which have structures affine with eIF-5A, CD83, B7 and / or CD40 receptors. This forms proteins that practically neutralize these receptors by forming complexes with B7 and / or CD40 receptors. In particular come to CTLA4, CD28, antibodies, F (ab) 2, scFv and / or F _ab fragments as proteins into account.

In a further preferred embodiment, the cell presenting antigen according to the invention contains nucleic acids which code for a signal sequence or expression products of a signal sequence which causes the expression products to remain in the endoplasmic reticulum, the Golgi apparatus, the Trans-Golgi network or intracellular vesicles.

To express antigens, the cell presenting the antigen according to the invention is transfected with nucleic acids which allow the expression of the expressed antigens in MHC II compartments of the cells.

All genetically modified antigen presenting cells present the autoantigen on most of their MHC complexes, while very few "normal" APCs present the autoantigen at all and then only on a few MHC complexes. The aim of the treatment is to displace the "normal" APCs which present the autoantigen and activate the T helper cells by the cells which have been programmed to switch off the antibody products and which present genetically manipulated antigen. It is useful to administer the antigen (s) in a form that allows transport to the MHC II endosomes. In this way, a presentation at MHC II can be reliably achieved. In the case of a genetic engineering intervention, this is usually done by manipulating the open reading frame, so that the reading frame is preceded by a signal sequence for this compartment. Genetic manipulation has the additional advantage that it has a much longer half-life than, for example, oligonucleotides or peptides. A stable integration of genes can even lead to a permanent change in the properties of the target cells. The corresponding nucleic acids can be DNA, RNA, oligonucleotides, polynucleotides, ribozymes, peptide nucleic acids (PNA).

The DNA preferably has regulatory elements such as enhancers, promoters, polyA-coding 3 ends for the transcription of the DNA into RNA, and the RNA regulatory elements for translating the RNA into protein. The regulatory elements ensure efficient expression of the genes.

The cell presenting the antigen according to the invention is produced, for example, by ex vivo or in vivo methods. An antigen-presenting cell is preferably transfected ex vivo or in vivo by treatment with viruses, viral vectors, bacterial vectors, plasmids, which are introduced into an antigen-presenting cell by electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules ,

In a further embodiment, an antigen-presenting cell or a monoantigenic antigen-presenting cell can be treated by treatment with viruses, viral vectors, bacterial vectors, plasmids by electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules into a cell with an increased amount of PD-1 binding molecules and / or with an increased amount of CTLA4 binding molecules and / or suppressed function of co-stimulatory receptors, CD83, eIF-5A or the expression of co-stimulatory receptors, CD83, eIF- 5A by preventing their expression or the co-stimulatory receptors, CD83, eIF-5A are prevented by stimulation with affine structures from stimulating T cells which are bound to the cell presenting the antigen according to the invention.

Different strategies can be used to suppress co-stimulation, CD83, eIF-5A, which ultimately means suppressing the production of one or more proteins or hindering the protein or proteins: 1. Antisense approach

2. Co-suppression

3. Binding of CD83, eIF-5A and additionally, if necessary, the co-stimulatory molecule. Re 1.: In this case, antisense nucleic acids must come into contact with the mRNA of the targeted molecule. You will then likely bind the molecule and prevent translation. A wide range of molecules, such as RNAs, DNAs, PNAs, ribozymes, are suitable as nucleic acids. Here, too, a genetic engineering intervention would ensure the longest half-life of the effect.

Re 2.: It has now been found that the production of a gene product can also be achieved by integrating a homologous sense gene sequence. The mechanism is still completely unknown. Re 3.: The binding of the targeted molecule, for example by specific antibodies, prevents this molecule from making contact with the targeted receptor on a T cell and thus prevents the activation of the T cell. The external addition of such binding molecules has the disadvantage that they act on all cells presenting antigen and thus prevent any immune reaction. In order to specifically hinder immune reactions, we have designed a model in which the targeted molecules are already bound in the cell, in an intracellular compartment. An extension of this model is intended to ensure that the binding molecule is retained in the intracellular compartment so that the targeted molecule does not reach the plasma membrane in the first place. Signal sequences are known for this, which must be added to the open reading frame of the binding molecule. This intracellular approach works well on isolated cells. Here, too, genetic engineering intervention is preferable. The desired effects can also be achieved if only one of the both goals are carried out with genetic engineering intervention. In this case, the following other molecules can be used instead of the genes:

1. Obstruction of the targeted molecules

Nucleic acids (mostly complementary to the target sequence), which are e.g. around oligonucleotides, polynucleotides, ribozymes, peptide nucleic acids

(PNAs) can act

Antibodies or other molecules that bind the targeted molecules.

2. Antigen contacting by proteins, peptides, peptidomimetics. In particular, molecules such as antibodies, proteins, peptides, peptidomimetics, PD-Ll, PD-L2, CTLA4, CD28, CD4ÖL and / or constituents and / or combinations of these molecules, which e.g. B7-1, B7-2, CD40 bind, which hinders co-stimulation of the T cell taking place in the presence of an antigen presentation, brought into contact with the mono-antigen presenting cell and / or the antigen presenting cell.

It can be advantageous to use molecules such as liposomes, hydrogels, cyclodextrins, nanocapsules, nanoparticles, in particular biodegradable nanocapsules or particles, bio-adhesive microspheres and / or by electroporation techniques, iontophoresis, ballistic methods and / or other techniques to transfer molecules into the monoantigenic antigen presenting cell and / or the antigen presenting cell.

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

A medicament containing at least one cell presenting an antigen according to the invention is furthermore claimed according to the invention. Vorzugswei- The medicament according to the invention is formulated as an infusion solution for intravenous or intraperitoneal application. The formulation is selected such that when the medicament is administered there is no significant impairment of the effectiveness of the cell presenting the antigen 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 quantities and pH values are of course not absolute, but can be varied and adapted to the specific needs of a patient by a specialist, depending on the conditions and requirements.

The cell presenting the antigen according to the invention can be used in particular for the production of a medicament for the treatment of unwanted immune reactions, such as autoimmune diseases and allergies or deliberately induced immune reactions, such as in immunizations.

Furthermore, the cell presenting the antigen according to the invention can be used for the production of a medicament for the treatment of immune reactions against allo-long and / or xenological tissue characteristics.

In particular, uses are claimed according to the invention in which the immune reactions to be treated are related to antigens or their gene sequences and / or parts thereof and are selected from the group consisting of - enzymes, their gene sequences and / or part sequences, in particular glutamic acid decarboxylase (GAD) , Receptor type protein tyrosine phosphatase IA-2Beta, antigen: H ⁺ K ⁺ ATPase, U1RNP, transglutaminase, argininosuccinate lyase (ASL), tyrosinase-related protein-2, thyroid peroxidase, factor VIII, factor IX; - Receptors, their gene sequences and / or partial sequences, in particular Nicotine-type acetylcholine receptor, bl-adrenergic receptor, al-adrenergic receptor, angiotensin-2-ATl receptor, glutamate receptor, thyrotropin-stimulating hormone (TSH) receptor, LFA-1, HLA-B27, epididymal protein DE, Zona Pellucida (ZP) -3 glycoprotein, Zona Pellucida (ZP) -4 glycoprotein, follicle-stimulating hormones (FSH) receptor, sperm

Immunogen SP-10 or Sperm Protein SP-10;

Hormones or messenger substances, their gene sequences and / or partial sequences, in particular insulin, thyroglobulin, follicle-stimulating hormones (FSH), prostaglandin F2 alpha, gonadotropin-releasing hormones (GnRH), estradiol-17beta, estrogen, luteinizing hormone (LH) receptor , Inhibin, testosterone, androgen, chorionic gonadotrophin (CG), interleukins, interferons, cytokines, chemokines, bone morphogenetic factors, b-interferon, estradiol;

- Structural proteins, their gene sequences and / or partial sequences, in particular myelin basic protein (MBP), proteolipid protein (PLP), myelin oligodendrocyte glycoprotein (MOG), a-fodrin, non-erythroid a-spectrin, beta-amyloid precursor Protein (beta-APP), type 2 collagen, sperm plasma membrane protein PH-20;

Antigens, their gene sequences and / or partial sequences, in particular CENP-A autoantigen, Beta2GP-I, ribosomal P protein, Ro / SSA, La / SSB,

Sm / RNP, Sm, Scl-70, Jo-1, BCOADC-E2, albumin, glucagon, islet cell antigens, Retinal S Ag;

Allergens that trigger an IgE response, their gene sequences and / or partial sequences, in particular Der f 1, Der f 2, Der f 3, Der p 1, Der p 2, Der p 3, Der p 4, Der p 5, The p 8, Eur m 1, Lep d 2, Fei d 1, Can f 1, Can f 2, Mus m 1, Rat n 1, Bla g 1, Bla g 2, Bla g 4, Bla g 5, Per a 1, bee venom phospholipase A2 (PLA2), Group V major allergenic Phl p 5b by Timothy Grass Pollen, Hom s 1.

Furthermore, the invention claims a use in which the immune reactions to be treated in connection with allologic and / or xenological tissue characteristics, their gene sequences and / or partial sequences, in particular special MHC I, MHC II, Rhesus factor. Development of a gene therapy

The method presented here for reducing immune responses, e.g. for the treatment of autoimmune diseases with a clearly defined autoantibody profile, is based on the genetic engineering manipulation of certain blood cells, preferably outside the body, which are then subsequently returned to the organism. The aim of this treatment is, among other things, the specific switching off of a chronic, immune system-driven production of autoantibodies that trigger the disease.

Areas of application of gene therapy

The procedure for the specific deactivation of immune reactions always makes sense where one or more molecularly defined targets (antigens, autoantigens) are known. These can be associated with an autoimmune disease or allergy, for example.

For example, are diseases with autoantibody-mediated autoimmune reactions, i.e. diseases in which the "binding structures" of these autoantibodies (autoantigens, targets) are known and have pathogenetic significance. Examples of diseases with known molecular structures (epitopes) that bind autoantibodies are:

• myasthenia gravis (autoantibodies against the acetylcholine receptor)

• Graves' disease (autoantibody against the TSH receptor of the thyroid gland)

Dilated cardiomyopathy (DCM, autoantibodies against the bl-adrenergic receptor of the heart muscle cells)

• certain forms of insulin-dependent diabetes (autoantibodies against insulin)

• certain forms of high blood pressure (autoantibodies against the angiotensin and / or al-adrenergic receptor). Treatment with genetically manipulated patient's own blood cells: principle of the proposed gene therapy

Within the immune system, the antibodies play an important role as defense molecules ("humoral immune response"). They are produced by mature B lymphocytes (B cells). However, the induction and production of the antibodies is not detached from the remaining cells of the immune system; rather, this humoral immune response is controlled by other cells in the immune system. The immune response cannot be maintained without the help and mediation of antigen presenting cells and T helper lymphocytes. The molecular mechanisms of the interaction - cell-cell contact - of antigen presenting cells with the T helper lymphocytes is known in detail at the receptor level.

In addition to the antigen to be presented, various receptors and auxiliary receptors are involved in cell-cell contact and cell activation on the cell surface. An essential molecule of the intercellular interaction in antigen presentation (contact of antigen presenting cells with the T helper lymphocytes) are costimulatory receptors with the names PD-Ll, PD-L2, B7h (LICOS), B7-1 and B7-2. CD40 also plays a role in this signal transduction. CD83 also has an influence on the stimulation ability of DCs.

The functions of ICOS, PD-1, CD28, CTLA4, B7 and CD40 are the contents of numerous publications (Daikh et al., 1997; Greenfield et al., 1998; Johnson-Leger et al., 1998; McAdam et al., 1998 ; Vyth-Dreese et al., 1995, (Freeman et al 2000; Hutloff et al 1999; Kopf et al 2000; Tamatani et al 2000)) and (apart from ICOS, PD1) are also extensively dealt with in textbooks [see. e.g. (Janeway and Travers, 1997)]. CD83 is also dealt with in the literature (Berchtold et al 1999; Kruse et al 2000a; Kruse et al 2000b). However, its exact functional mechanisms are still largely unknown.

The designed procedure is based on up to two parallel interventions on the patient's own antigen presenting cells. The interventions are carried out using suitable probes and have an effect • Shutdown of CD83 and / or eIF-5A, which prevents DCs from activating T cells

And optionally the production of a PD-1 binding molecule, e.g. PD-Ll, which via the action of PD-1, which occurs in T cells, prevents the T cells from activation and proliferation

And optionally the production of a CTLA4 binding molecule, e.g. Antibodies that prevent the T cells from activating through the action of CTLA4, which occurs in T cells

• and possibly switching off B7 and thus preventing the formation of the molecule on the surface of the antigen presenting

Cells and / or the shutdown of CD40

• and, if necessary, at the same time a strong presentation of the autoantigen (s) on the antigen presenting cells

If the cells that are antigen presenting cells are manipulated ex vivo, the cells are returned to the donor's bloodstream. The cells presenting the genetically manipulated antigen now switch off the corresponding T helper lymphocytes in the organism. The genetically manipulated cells compete directly with the APCs that are already present in the organism (preferably the bloodstream and lymphatic system), but which carry B7 and no additional PD-1 binding molecule, CTLA4 binding molecule on the cell surface and when they mature DCs are, have CD83 on the plasma membrane and normally activate the T helper lymphocytes and thus, among other things, antibody production. Treatment of autoimmune diseases, allergies and graft rejection reactions: state of the art

Causal therapies for the treatment of autoimmune diseases and allergies do not exist. With a few exceptions (extracorporeal elimination of antibodies from the patient's blood or plasmapheresis), autoimmune diseases and allergies are treated with medication by inhibiting immune system reactions.

The most common is the medicinal treatment of autoimmune diseases, allergies and graft rejection reactions with immunosuppressive preparations such as cortisone and its derivatives, as well as cyclosporin, beta-interferon or cytostatics (methothrexate). Furthermore, antibodies, antagonists and oligonucleotides against various signal components of the immune system are being tested with the purpose of preventing the activation of T cells. Such drugs are at best selective, but never specifically directed against the wrongly programmed autoimmune reactions. Immunosuppressants therefore have a negative effect on important, necessary and useful parts of the immune system; for this reason and their side effects, their use is limited.

The concept of gene therapy for the reduction of specific immune reactions differs fundamentally from immunosuppressive therapies. A specific shutdown of misdirected immunological reactions by genetically reprogrammed patient's own blood cells is carried out.

The methodology shown is intended as a generally applicable concept for reducing immune responses. With this basic concept it should be possible to stop any immune reaction of the adaptable immune system. In addition, such immune reactions can be triggered at will and then switched off again.

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Claims

claims

1. Antigen presenting cell (APC), characterized in that in the APC the function of CD83 and / or the eukaryotic initiation factor 5A (eIF-5A) and optionally one of the functions of co-stimulatory receptors, such as B7 and optionally CD40 receptors are suppressed and if necessary a CTLA4 binding molecule is produced and if necessary a PD-1 binding molecule is produced.

2. Antigen presenting cell according to claim 1, characterized in that the antigen presenting cell is a monocyte, a dendritic cell, a B cell (B lymphocyte) and / or a macrophage.

3. Antigen presenting cell according to claim 1 and 2, characterized in that the antigen presenting cell predominantly presents certain antigens previously (monoantigenic antigen presenting cell).

4. Monoantigenic antigen presenting cell according to claim 3, characterized in that by transfecting an antigen presenting cell with nucleic acid-containing material there is an increased expression of the predefined antigens and the antigen presenting cell essentially presents only these antigens.

5. Antigen presenting cell according to at least one of claims 1 to 4, characterized in that the antigen presenting cell according to the invention has an increased number of homing receptors, such as CD44.

6. Antigen presenting cell according to at least one of claims 1 to 5, characterized in that a transfection is a suppression of

Functions of the CD83, eIF-5A molecules, B7, CD40 receptors and / or the expression of CTLA4 binding molecules and / or an expression of PD-1 binding molecules and possibly an increase in the number of homing receptors.

7. Antigen presenting cell according to at least one of claims 1 to 6 containing nucleic acids encoding molecules that bind PD-1 and / or encoding molecules that bind CTLA4 and / or antisense nucleic acids to prevent CD83, eIF-5A, B7 and / or CD40 receptor expression.

8. Antigen presenting cell according to at least one of claims 1 to 7 containing nucleic acids, which suppress the expression of the CD83 molecules, eIF-5A, B7 and / or CD40 receptors by co-suppression.

9. antigen presenting cell according to at least one of claims 1 to 8 in which the PD-1 binding molecules are preferably PD-L1, PD-L2, antibodies, monoclonal antibodies, single chain antibodies (scFv).

10. Antigen presenting cell according to at least one of claims 1 to 9 containing nucleic acids which code for PD-1 binding molecules which remain in the plasma membrane of the cell.

11. Antigen presenting cell according to at least one of claims 1 to 10 containing nucleic acids which bring about an expression of proteins or peptides having structures affinity with CD83 molecules, eIF-5A, B7 and / or CD40 receptors.

12. The antigen-presenting cell according to claim 11, wherein the proteins which have structures affecting CD83 molecules are antibodies, F (ab) 2, scFv and / or F _a b ^_ fragments.

13. The antigen presenting cell according to claim 11, wherein the proteins which have structures affine to eIF-5A are antibodies, F (ab) 2, scFv and / or Fab fragments.

14. The antigen presenting cell according to claim 11, wherein the proteins which have structures affine to B7 receptors are CTLA4, CTLA4 derivatives (for example CTLA4Ig), CD28, antibodies, F (ab) 2, scFv and / or F _a b fragments are.

15. Antigen presenting cell according to claim 11 and / or 12 and / or 14, wherein the nucleic acids code for a signal sequence or the express sion products 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.

16. Antigen presenting cell according to at least one of claims 3 to 15, characterized in that the antigen presenting cell according to the invention is transfected for expression of antigens with nucleic acids, which enables the transport of the expressed antigens in MHC II compartments of the cells.

17. Antigen presenting cell according to at least one of claims 3 to 16, characterized in that the nucleic acids are DNA, RNA, oligonucleotides, polynucleotides, ribozymes, peptide nucleic acids (PNA).

18. Antigen presenting cell according to claim 17, characterized in that the DNA contains regulatory elements such as enhancers, promoters, polyA-coding 3 'ends for transcription of the DNA into RNA, which contains RNA regulatory elements for translating the RNA into protein.

19. A method for producing the antigen presenting cell according to at least one of claims 1 to 18 by ex vivo or in vivo methods.

20. The method of claim 19, wherein an antigen presenting cell ex vivo or in vivo by treatment with viruses, viral vectors, bacterial vectors, plasmids by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods and / or other techniques for infiltration is transfected by molecules into an antigen presenting cell.

21. The method according to claim 19 and / or 20, wherein an antigen presenting cell or a monoantigenic antigen presenting cell

Treatment with viruses, viral vectors, bacterial vectors, plasmids which bind by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules into a cell with increased production of PD-1 binding molecules and / or CTLA4 Molecules and / or with suppressed Function of co-stimulatory receptors, CD83, eIF-5A molecules, the expression of co-stimulatory receptors, CD83, eIF-5A molecules by preventing their expression or the co-stimulatory receptors, CD83, eIF-5A Molecules are prevented from reacting with affine structures to stimulate T cells bound to the antigen presenting cell or monoantigenic antigen presenting cell.

22. The method according to at least one of claims 19 to 21, wherein molecules such as antibodies, proteins, peptides, peptidomimetics, PD-Ll, PD-L2, PD-1 binding molecules, CTLA4 binding molecules, CTLA4, CTLA4 derivatives

(e.g. CTLA4Ig), CD28, CD40L and / or constituents and / or combinations of these molecules which e.g. PD-1, CTLA4, B7-1, B7-2, CD40, which prevents stimulation and / or co-stimulation of the T cell taking place in the presence of an antigen presentation, with the monoantigenic antigen presenting cell or the antigen presenting cell in

Be brought in contact.

23. The method according to at least one of claims 19 to 22, wherein the molecules according to claim 20 by vehicles such as liposomes, hydrogels, cyclodextrins, nanocapsules, nanoparticles, bio-adhesive microspheres and / or by electroporation techniques, iontophoresis, ballistic methods and / or others Techniques for introducing molecules into the monoantigenic antigen presenting cell or the antigen presenting cell are transferred.

24. The method according to at least one of claims 19 to 22, wherein nucleic acids by viruses, viral vectors, bacterial vectors, plasmids by viral gene transfer, electroporation techniques, iontophoresis, ballistic methods and / or other techniques for introducing molecules into the monoantigenic antigen presenting cell or the antigen presenting cell.

25. Antigen presenting cell according to at least one of claims 1 to 24, characterized in that an inhibitor of hypusin biosynthesis, such as GC7 (N ¹ -guanyl-l, 7-diaminoheptane) is used to hinder the production of functional eIF-5A.

26. Medicament containing at least one antigen presenting cell or monoantigenic antigen presenting cell according to at least one of claims 1 to 18.

27. Medicament according to claim 26, wherein at least one antigen presenting cell or monoantigenic antigen presenting cell is formulated as an infusion solution for intravenous or intraperitoneal application.

28. Use of at least one antigen-presenting cell or mono-antigenic antigen-presenting cell according to at least one of claims 1 to 18 for the manufacture of a medicament for the treatment of unwanted immune reactions, such as autoimmune diseases and allergies or deliberately induced immune reactions, such as in immunizations.

29. Use of at least one antigen presenting cell or monoantigenic antigen presenting cell according to at least one of claims 1 to 18 for the manufacture of a medicament for the treatment of immune reactions against allologic and / or xenological tissue characteristics.

30. Use according to claim 28, wherein the immune reactions to be treated are associated with antigens or their gene sequences and / or parts thereof are selected from the group consisting of

- Enzymes, their gene sequences and / or partial sequences, in particular glutamic acid decarboxylase (GAD), receptor-type protein tyrosine

Phosphatase IA-2Beta, H ⁺ K ⁺ ATPase, U1RNP, transglutaminase, argininosuccinatease (ASL), tyrosinase-related protein-2, thyroid peroxidase, factor VIII, factor IX;

- Receptors, their gene sequences and / or partial sequences, in particular acetylcholine receptor of the nicotine type, myasthenia gravis, bl-adrenergic receptor, al-adrenergic receptor, angiotensin-2-ATI receptor, glutamate receptor, thyrotropin-stimulating hormone (TSH) receptor , LFA-1, HLA-B27, Epididymal Protein DE, Zona Pellucida (ZP) -3 glycoprotein, Zona Pellucida (ZP) -4 glycoprotein, Follicle-Stimulating Hormone (FSH) receptor, Sperm Immunogen SP-10 or Sperm Protein SP-10;

Hormones or messenger substances, their gene sequences and / or partial sequences, in particular insulin, thyroglobulin, follicle-stimulating hormones

(FSH), prostaglandin F2 alpha, gonadotropin-releasing hormones (GnRH), oestradiol-17beta, estrogen, luteinizing hormone (LH) receptor, inhibin, testosterone, androgen, chorionic gonadotrophin (CG), interleukins, interferons, cytokines, chemokines, bones Morphogenetic factors, b-interferon, estradiol;

- Structural proteins, their gene sequences and / or partial sequences, in particular myelin basic protein (MBP), proteolipid protein (PLP), myelin oligodendrocyte glycoprotein (MOG), a-fodrin, non-erythroid a-spectrin, beta-amyloid precursor protein ( beta-APP), type 2 collagen, sperm plasma membrane protein PH-20;

Antigens, their gene sequences and / or partial sequences, in particular CENP- A autoantigen, Beta2GP-I, ribosomal P protein, Ro / SSA, La / SSB, Sm / RNP, Sm, Scl-70, Jo-1, BCOADC-E2, Albumin, glucagon, islet cell antigens, Retinal S Ag; Allergens that trigger an IgE response, their gene sequences and / or partial sequences, in particular Der f 1, Der f 2, Der f 3, Der p 1, Der p 2, Der p 3, Der p 4, Der p 5, The p 8, Eur m 1, Lep d 2, Fei d 1, Can f 1, Can f 2, Mus m 1, Rat n 1, Bla g 1, Bla g 2, Bla g 4, Bla g 5, Per a 1, bee venom phospholipase A2 (PLA2), Group V major allergenic Phl p 5b by Timothy Grass Pollen, Hom s 1.

31. Use according to claim 29, wherein the immune reactions to be treated are related to allologic and / or xenologic tissue characteristics, their gene sequences, and / or partial sequences, in particular MHC I, MHC II, rhesus factor.