EP3440464A1

EP3440464A1 - Method and device for the extracorporeal removal of pathogens and/or an excess of components from a cell sample of a patient

Info

Publication number: EP3440464A1
Application number: EP17718824.0A
Authority: EP
Inventors: Walter Schubert
Original assignee: Toposnomos Ltd
Current assignee: Toposnomos Ltd
Priority date: 2016-04-08
Filing date: 2017-04-04
Publication date: 2019-02-13
Also published as: CN109791153A; WO2017174188A1; DE202017007621U1; US20210008271A1; DE102016106510A1

Abstract

The invention relates to a method for the extracorporeal removal of pathogens and/or an excess of components from a cell sample of a human or animal patient suffering from an illness. The method comprises the steps: a) determination of at least one protein cluster (CMP) that is characteristic of the illness of the patient; b) preparation of the cell sample of the patient; and c) extracorporeal removal of components having the at least one determined protein cluster from the cell sample. The invention further relates to a device for carrying out said method.

Description

Method and device for extracorporeal removal of pathogenic and / or supernumerary components from a cell sample of a patient

description

The invention relates to a method and a device for extracorporeal removal of pathogenic and / or excess components from a cell sample of a human or animal patient. The corporeal treatment of many diseases is often associated with severe side effects or even impossible so far. Alternatively, devices are known which are located outside the body (extracorporeal) of the patient and with the aid of which cell samples of the patient can be treated and subsequently returned. As a result, in some cases, a gentler treatment of the disease in question is possible because, for example, the immune system of the patient is affected to a lesser extent by the treatment than would be possible by ancorporeal treatment.

It is to be regarded as disadvantageous that the known extracorporeal treatment methods and the devices used to carry them out are comparatively unspecific and therefore only have relatively fewer illnesses for corresponding non-specific treatment.

The object of the present invention is to provide a method and a device for extracorporeal removal of pathogenic and / or excess components from a cell sample of a human or animal patient, which allow a more specific and flexible treatment of a variety of diseases. The objects are achieved by a method with the features of claim 1 and by a device according to claim 16. Advantageous embodiments with expedient developments of the invention are specified in the respective subclaims, wherein advantageous embodiments of the method are to be regarded as advantageous embodiments of the device and vice versa.

A first aspect of the invention relates to a method for the extracorporeal removal of pathogenic and / or supernumerary components from a cell sample of a human or animal patient suffering from a disease, in which at least the steps a) determining at least one protein cluster (CMP) suitable for characterizing the disease of the patient; b) providing the patient's cell sample; and c) extracorporeal removal of constituents having the at least one detected CMP from the cell sample. As a result, a large number of diseases can be treated in a highly specific and low-side-effect manner, since only the affected components of the cell sample are removed outside the patient's body, whereas cells of the cell sample not affected by the disease are not removed or at least remain largely unchanged. The term "removal" in the context of the present invention encompasses not only a partial or complete "removal" of the relevant components, but alternatively or additionally also a partial or complete inactivation or destruction of these components, whereby these at least no longer in their pathogenic form in the cell sample present or be depleted as much as possible.

Within the context of the present invention, a protein cluster is understood to mean a combinatorial molecular phenotype (CMP), the colocalized and / or anti-colonized CDs (clusters of differentiation, groups), proteins and / or molecules in a particular location Cell describes. Accordingly, groups of CMPs represent regions of colocalized and / or anti-colonized CDs, proteins, and / or other molecules in a cell or cell complex of the cell sample. For example, a CMP in this context may be given in the form of a binary code, where it is possible to encode each digit of the binary number (ie, each "bit") to determine whether a predetermined CD / protein / molecule is within a given one Location and / or in a concentration exceeding a certain limit (eg L, 1 or True) occurs in the cell. In the other case, that is, if the predetermined CD / protein / molecule is not present at this point of the cell. occurs or only in a below a threshold concentration, a different coding (z, B. Ä, 0 or False) is used. Likewise, it may be provided that certain CDs / proteins / molecules are only occasionally colocalized with one or more other CDs / proteins / molecules and sometimes are not colocalized. These CDs / proteins / molecules can be encoded with "wildcards" (eg W, ^* ).

The invention is based on the finding that certain CDs / proteins / molecules function as disease-specific and / or patient-specific as so-called lead proteins. At a high level of molecular cell organization, proteins are often coupled together as clusters or networks (corresponding to said CMP motifs), which is controlled by lead proteins: when the lead proteins are inhibited or removed, these clusters disintegrate, causing their loss biological function leads. Therefore, many diseases can be treated effectively and at least essentially without side effects by determining the CMP or group of CMPs that is characteristic of a particular disease, according to which the particular constituents of the cell sample carrying these pathological "markers" , be removed extracorporeally from the cell sample or inactivated.

Depending on the disease and the success of the treatment, the method according to the invention can in principle be carried out once or several times. For example, multiple cycles may be performed at certain consistent or varying intervals. The number and frequency of the procedures should be determined and controlled by a physician.

It has proven to be advantageous if a body fluid, in particular blood and / or blood plasma, and / or a tissue sample of the patient is provided as the cell sample. In this way, the cell sample can be optimally selected depending on the disease to be treated.

Further advantages result from the at least one CMP being based on a database and / or by means of a multi-epitope ligand cartography (MELK) or ICM method (multi-epitope ligand cartography / imaging cycler microscopy) and / or is determined by means of a MELK robot system. For diseases in which the characteristic CMP or the characteristic group of CMPs is already known and not usually patient-specific, it may be sufficient to determine the CMP (s) from a corresponding medical database. Alternatively or additionally, it may be provided that the CMP (s) characteristic for the disease is / are determined with the aid of a multi-epitope ligand cartography (MELK) or I CM method (imaging cycler microscopy) and / or is determined by means of a MELK robot system or be. The MELK / IC M method or the MELK robot system are as such, for example, from the documents US Pat. No. 6,150,173, DE 197 09 348 A1, EP 0 810 428 A1, DE 100 43 470 A1 and WO

02/21 137 A2 known.

Alternatively or additionally, it is provided that the extracorporeal removal of the components is monitored and / or controlled by means of a MELK or IC method and / or by means of a MELK robot system. In this way, the desired depletion of the disease-specific components of the cell sample can be controlled and optionally controlled or regulated.

A particularly specific and therefore effective removal of the pathogenic and / or supernumerary components from the cell sample is possible in a further embodiment of the invention if the disease is a tumor disease and / or an inflammatory disease. Such diseases have a particularly high degree of cellular organization and can accordingly be specifically addressed in the context of the present invention.

More valuable advantages are achieved by carrying out the extracorporeal removal by means of an apheresis method and / or by means of an apheresis device. In particular, pathogenic and / or supernumerary constituents, which are found at least predominantly in the blood and / or blood plasma of the diseased patient, can thereby be removed from the blood and / or blood plasma particularly effectively and with little side effects.

It has also been shown to be advantageous if, as apheresis method, an unselective apheresis and / or a selective apheresis and / or a whole blood aphrodisiac Rese and / or a photopheresis is performed and / or that the Apheresevor- device for performing at least one apheresis method from the group unselective apheresis, selective apheresis, whole blood apheresis and photopheresis is formed. This allows an optimal adaptation to the respective disease or to the characteristic of the disease components. In extracorporeal apheresis, in some embodiments, the patient's blood may first be separated by means of a centrifuge to enrich the pathogenic components for non-pathogenic components. It is also possible to completely run and / or substitute certain components. In the context of a photopheresis, the pathogenic constituents are subjected to a controlled irradiation with light of a certain wavelength (s) in order to achieve destruction or depletion. The wavelength (s), irradiation time and irradiation time can be selected depending on the properties of the pathogenic components. For example, the optionally separated or fractionated cell sample can be exposed to controlled UV-A and / or UV-B irradiation. Following apheresis, the blood or blood plasma in which the pathogenic components have been depleted or completely removed may be returned to the patient. The treated cell sample is thereby presented to the untreated immune system and in some cases may cause additional modulation of the immune system.

In a further advantageous embodiment of the invention, it is provided that for extracorporeal removal at least one streptamer and / or at least one antibody and / or at least one ligand, in particular a variable single-chain fragment (scFv) and / or a multivalent antibody fragment (scFv multimer) , is used. As a result, a selection optimally adapted to the component to be removed can be carried out. The at least one streptamer and / or the at least one antibody and / or the at least one ligand can in principle be added to the cell sample and / or used immobilized on a carrier material. Antibody fragments offer the advantage of high binding affinity and specificity for a wide range of targets and haptens. Single-chain fragments can also be cross-linked or expressed as diabodies (60 kDa), triabodies (90 kDa), tetrabodies (120 kDa), etc., whereby different linker lengths between V domains are possible. A special Another advantage is that molecules of 80-120 kDa increase the penetration of cells and have faster clearance rates than corresponding igs (150 kDa). It may further be provided that the antibody and / or ligand is a diabody, triabody, tetrabody, pentabody, hexabody, heptabody or octabody. In other words, it is provided that the antibody and / or ligand mono-, bk tri-, is formed quad, pent, hex, hept, oct, enn- or multi-specific. This allows the cross-linking of two, three, four, five, six, seven or eight target structures or target proteins, whereby scFv multimers can be adapted particularly precisely and individually to the possibly patient-specific spatial arrangement of the target epitopes of specific combinatorial protein clusters (CMPs). The increased binding value of scFv multimers leads to a particularly high activity.

More valuable advantages result from the fact that the at least one streptamer and / or the at least one antibody and / or the at least one ligand specifically binds and / or inactivates a lead protein that is characteristic of the disease. In this way, a particularly efficient and comprehensive removal of the pathogenic and / or excess components can be achieved and a correspondingly reliable treatment of the disease in question can be made possible.

In a further advantageous embodiment of the invention, it is provided that the disease is amyotrophic lateral sclerosis (ALS) and / or that the cell sample is a blood sample of the patient and / or that the at least one CMP one or more from the group CD16, CD8, NeuN , Bax, Bcl2, CD1 1 b, CD138, CD16A, CD29, CD2, CD45RA, CD49d, CD54, CD56, CD57, CD58, CD62L, CD3, HLADR, immunoglobulin G, MHCII, MHCI, SIRT1, RAC1, BMX, GAK, JNK2, APKK6, OTUB2, PRKAR2A, SMAD2, SMAD4 and STAP2 comprises and / or that the components by means of at least one antibody and / or ligand, which is at least one of the group CD16, CD8, NeuN, Bax, Bcl2, CD1 1 b, CD138 , CD16A, CD29, CD2, CD45RA, CD49d. CD54, CD56, CD57, CD58, CD62L, CD3, HLADR, immunoglobulin G, MHCII, MHCI, SIRT1, RAC1, BMX, GAK, JNK2, MAPKK6, OTUB2, PRKAR2A, SMAD2, SMAD4 and STAP2 bind, and / or by means of a CD16 -Ectodomain shedding molecule can be removed extracorporeally. Amyotrophic lateral sclerosis (ALS) relies on an unstoppable and rapid progression irreversible paralysis of the musculature, because certain nerve cells, which control the musculature, degenerate. So far, more than 50 clinical treatment studies have been carried out, but their treatment concepts have proven largely ineffective. With the aid of the method according to the invention, it is possible for the first time to remove or inhibit the pathogenic ALS-specific cells extracorporeally, as a result of which the progression of the disease can be stopped or at least considerably slowed down. The pathogenic components responsible for the ALS are aberrant T-lymphocytes which, in addition to the CD8 receptor, also express the CD16 receptor complex. Therefore, they are characterized by CMPs containing CD8 and / or CD16 as lead proteins. The lead proteins can be colocalized or anti-colonized with the other indicated CDs and proteins. Accordingly, it has been found to be advantageous if the antibody and / or ligand used to remove the pathogenic constituents contains one, two, three, four, five or more of the group CD16, CD8, STTP1, NeuN, Bax, Bcl2, CD1 1b , CD138, CD16A, CD29, CD2, CD45RA, CD49d, CD54, CD56, CD57, CD58, CD62L, CD3, HLADR, immunoglobulin G, MHCII, MHCI, SIRT1, RAC1. BMX, GAK, JNK2. MAPKK6, OTUB2, PRKAR2A, SMAD2, SMAD4 and STAP2 binds. In other words, it is provided that the antibody and / or ligand mono-, bk tri-, quad, pent, hex, hept, oct, denominated or multi-specific. These proteins, in various combinations, form supra-individual and individual clusters in ALS-specific cells and can therefore be cross-linked extracorporeally and thus blocked, as a result of which the ALS-specific cells lose their functionality. This makes it possible to treat ALS with a particularly specific and thus low-side-effect or even side effect-free treatment. Further advantages arise when the antibody and / or ligand binds at least CD16, CD8 and STTP1. STTP1 here denotes the possibly patient-specific signal protein (signal transduction p red in 1, eg kinase) which mediates the signal cascade extending from the cell surface to the nucleus and together with the module "CD16a and CD8" at the Surface of the cell where it is colocalized with CD16a and CD8 STTP1 can be prepared using the well-known MELK and / or ICM technique (Multi Epitope Ligand Cartography or Imaging Cycler Microscopy) for the respective patient It may further be provided that STTP1 is at least one protein from the group RAC, STAP2 and SMAD2 when the antibody and / or ligand is recombinant, human or de novo.

Further advantages are obtained by using an antibody and / or ligand for extracorporeal removal, which binds two or more from the group CD16, CD8, RAC1, STAP2 and SMAD2. This may block ALS-specific protein clusters which, in addition to one or more cell surface proteins (eg CD8, CD16, CD45RA) are additionally directly associated with one or more signal chain molecules from the group RAC1, STAP2 and / or SMAD2, RAC1 (Ras-related C3 botulinum toxin substrate 1) as a member of the RAG subfamily regulates a variety of cellular events including cell growth control, cytoskeletal reorganization, and activation of protein kinases. STAP2 (signal transducing adapter p red in 2) and SMAD 2 (mothers against decapentaplegic homolog 2) are also part of the signal transduction chain and regulate the signal transduction and transcription of central signaling pathways in ALS-specific cells.

Alternatively, it is provided that the disease is prostate cancer and / or that the cell sample is prostate tissue and / or that the at least one CMP comprises one or more of the group CD26 and CD29 and / or that the components are at least one antibody and / or or ligands which bind at least one of the group CD26 and CD29 are removed extracorporeally. This makes it possible for the first time to treat prostate cancer extracorporeally and to monitor the therapy. The invention is based on the finding that prostate cancer is characterized by CMPs which contain CD26 and / or CD26 as lead proteins, which are optionally colocalized with other CDs or proteins or are antico-localized.

Further advantages result if the at least one CMP also comprises CD44 and / or CD54 and / or CD138 and / or if the at least one CMP has at least one of CD3, CD4, CDS, CD10, CD13, CD19, CD20, CD38, CD49d, CD58 and CD80 are missing. This allows a particularly reliable identification and monitoring of the removal of the pathogenic components of the cell sample which are responsible for prostate cancer. Prostate cancer is particularly present when the cell and / or when at least 70%, preferably at least 75% of the CMPs on the cell surface of at least one cell contain CD26 and / or CD29, and / or if at least 20%, preferably at least 35% of the CMPs on the cell surface of at least one cell comprise CD28 and / or CD29 and they lack at least CD3, CD4, CD8, CD10, CD13, CD19, CD20, CD38, CD49d, CD58 and CD80.

Accordingly, in a further embodiment of the invention, the pathogenic constituents of the cell sample can be removed particularly reliably by extracorporeal removal using an antibody and / or ligand which binds at least one or more of CD26, CD29, CD44, CD54 and CD138. Alternatively, it is provided that the disease is a cutaneous lymphoma, in particular mycosis fungoides, and / or that the cell sample is a skin sample and / or that the at least one CMP is one or more of the group HLA-DQ, CD2, CD3, CD4, CD7 , CDS, CD10, CD13, CD18, CD18, CD26, CD29, CD36, CD44, CD45, CD49f, CD54, CD56, CD57, CD58, CD62L, CD71, CD80 and HLA-DR and / or that the constituents are at least one antibody and / or ligand comprising at least one of the group HLA-DQ, CD2, CD3, CD4, CD7, CDS, CD10, CD13, CD18, CD18, CD26, CD29, CD36, CD44, CD45, CD49f, CD54, CD56, CD57, CD58, CD62L, CD71, CD80 and HLA-DR binds to be removed extracorporeally. This makes extracorporeal treatment and therapy control of cutaneous lymphomas possible for the first time. The invention is based on the finding that cutaneous lymphomas are characterized by CMPs, which in particular contain HLA-DQ as lead protein, which is colocalized or antico-localized with one or more of the said CDs or proteins. In a further advantageous embodiment of the invention, the cell sample is returned to the patient after the partial or complete extracorporeal removal of the constituents which have the at least one determined CMP. A second aspect of the invention relates to a device which is designed to carry out a method according to the first aspect of the invention. In the context of the present invention, the term "trained to be" means an apparatus which not only has a fundamental suitability for carrying out such a method, but is actually set up and equipped to carry out such a method. Features and their advantages can be found in the descriptions of the first aspect of the invention, wherein advantageous embodiments of the first aspect of the invention are to be regarded as advantageous embodiments of the second aspect of the invention and vice versa.

Further advantages result from the device being designed to carry out at least one apheresis method from the group of unselective apheresis, selective Aphere se, full blood apheresis and photopheresis. In the case of the non-selective aphe- sis, which is also referred to as plasma exchange, the plasma is separated from the blood by means of the device and completely substituted. Mostly human blood products are used as replacement fluids. In selective plasmapheresis (plasma perfusion), the cells carrying the protein cluster (CMP), which is characteristic of the disease of the patient, are separated from the plasma by filtration or adsorption by means of the device. The purified plasma can then be returned to the patient. In whole blood apheresis (hemofusion), the cells which carry the protein cluster (CMP), which is characteristic of the disease of the patient, for example, are filtered affinity chromatography directly from the blood by means of the device. In the case of extracorporeal photopheresis, the cell sample or the blood of the patient can first be separated by means of a centrifuge of the device, leukocyte-enriched blood plasma can be collected by means of the device and exposed to the device in an extracorporeal cycle of controlled UV irradiation by means of a UV irradiation device , The wavelength of the UV irradiation is typically between 340 and 380 nanometers, for example 365 nanometers (UV-A). The average UV exposure of the leukocytes can typically be between 0.1 and 5 J / cm ² , for example 1.5 J / cm ² . The total duration of the treatment is between 2 and 5 hours, depending on the volume of the cell or blood sample. The sample can be used in extracorporeal photopheresis by means of the device a photoactivatable pharmaceutical, for example methoxsalen, may be added. Following the irradiation, the cell sample or the blood can be returned to the patient partially or completely. The advantage is that the immune system of the patient is not or only to a limited extent mitigated, so that in addition to the direct cytostatic and cytotoxic effects on the irradiated cell or blood sample, an immune modulation of the non-irradiated immune system can be effected. The treatment frequency can be 4 to 20 cycles in 2-20 day intervals. The intervals between individual treatments may increase over time. Treatment success can be achieved by determining the decrease in the concentration of cells that make up the protein cluster

(CMP), which is characteristic of the disease of the patient Another advantage of apheresis is the relatively low cost. According to an Australian analysis, the cost per ALS patient and year currently amounts to about 750,000 to 1 million euros. For example, with a topical treatment ALS can at least stop in early stages, with up to about 26 treatments per year, which can be realized for a fraction of the previous annual costs, the emergence of progressive severe disability and the associated burden can be prevented which, in addition to the unquantifiable gain in lifetime and quality of life for the patient, also leads to significant savings in healthcare.

A third aspect of the invention relates to a streptamer and / or an antibody and / or a ligand for use in a method according to the first aspect of the invention. The resulting features and their advantages are described in the descriptions of the first aspect of the invention, with advantageous

Embodiments of the first aspect of the invention are to be regarded as advantageous embodiments of the third aspect of the invention and vice versa. The streptamer, the antibody and / or the ligand may be coupled in a further embodiment with an adsorber and / or a chromophore. As a result, they are particularly suitable for use in the context of a photopheresis with tuned to the absorber or the chromophore wavelengths. Alternatively or additionally, the streptamer, the antibody and / or the ligand can be coupled with a marker, in particular a magnetic marker (magnetic cell separation). This allows a simple extracorporeal separation of with the Strepta- mer / antibody / ligand bound pathogenic components by affinity chromatography, for example with the aid of so-called MicroBeads. These are about 50 nanometers large magnetic particles that are bound to the streptamer, the antibody or the ligand. Streptamer / antibody / ligand recognize disease-specific CMP (s) on the surface of pathogenic components (cells) and thus ensure the binding of the MicroBeads to this cell population. As the cell sample flows through a column surrounded by a strong magnetic field, the cells labeled with the MicroBeads are retained. In this way, when rinsing the column, only unlabelled and therefore non-pathogenic cells are obtained, so that the marked cell population and thus the constituents characteristic of the disease were removed from the original cell mixture. After removal of the magnetic field also the marked cell population can be obtained by rinsing the column and is thus available for further experiments. By repeated treatment of a cell mixture with different MicroBeads also a so-called fractionated separation, ie the separation into several cell populations, is possible. In a further embodiment, the streptamer, the antibody and / or the ligand can be coupled to a fluorescent dye, a quantum dot, a radioactive marker, a spin label and / or a tag for a further antibody / ligand and / or an enzyme ,

Further features of the invention will become apparent from the claims, the figures and the description of the figures. The features and combinations of features mentioned above in the description, as well as the features and feature combinations mentioned below in the description of the figures and / or shown alone in the figures, can be used not only in the respectively indicated combination but also in other combinations without the scope of the invention leave. There are thus also embodiments of the invention as encompassed and disclosed, which are not explicitly shown and explained in the figures, however, emerge and can be generated by separated Merkmaiskombinationen from the described embodiments. Embodiments and combinations of features are also to be regarded as disclosed, which thus do not have all the features of an originally formulated independent claim. Showing: Ia to If different topographical images as more specific, abnormal

cells; Figures 2a to 2f show the step process of these abnormal cells leading to the ALS specific disease;

Fig. 3 is a schematic representation of the topological hierarchy of

Proteins within a toponome;

Fig. 4 is a Toponomkarte, the location of the 30 most of more than

Shows 2,000 different CMPs in prostate cancer;

Figure 5 is a representation of the selective expression of the prostate cancer-specific CMP in subcellular sites of neoplastic

Prostate acini;

Fig. 6 is an illustration of the cyclic localization of 25 biomolecules and the subsequent determination of the CMPs using the MELK / ICM method;

Fig. 7 Toponomabbildungen the tissue organization of a cutaneous

lymphoma; Fig. 8 is a schematic representation of the pathomechanism of

WHEN; and

Fig. 9 is a schematic representation of a therapy of ALS. Mechanisms of ALS in the toponom

Synopsis of the ALS Toponome: Analyzes of cell surfaces of immune cells in the blood have shown that ALS has a specific abnormal topon. Essential feature is the existence of aberrant T-lymphocytes, in addition to also express the CD16 receptor complex to the CD8 receptor. Exactly the same cell forms are found in the morphologically well-preserved ALS postmortem tissue within postcapillary venules of the spinal pyramidal tract (Figure 1 a, b; Box). Detailed 3D topon resolution indicates that these cells express the CD16 complex (Figure 5 1, arrow 1), are polarized and show a "leading edge" with CD3 positive membrane penetrating the endothelial cell layer of the blood vessel (Figure 1c; Arrow 2), while CD3 positive vesicles are transported intracellularly to the "leading edge" (Figure 1 c, box). Individual vesicles contain complexes of CD8 / CD16 (Figure 1 d, e, f). These CD8 / CD16 complexes are required by "forward transport" for transient migration of these cells through the endothelial cell layer via vesicle budding in the cell surface to promote transmigration as part of an abnormal "homing code" CD16 is also referred to as Fe region reeeptor III-A or Fcylll After transmigration, this complex is degraded because CD8 positive T cells in the parenchyma of the pyramidal tract do not have the CD16 complex 15. Instead, they are found as single cells between the myeli - Neurons (Figure 2 a, CD8 + CD16 - blue cell) Increased 3D magnification of this cell has a cell process that has penetrated deeply into the surface of a morphologically intact axon (Figure 2 b, Box; c: Enlargement of the box: star = morphologically intact axon body.) This cell process expresses CD3 and 0 CD8 (Figure 2, arrows 1, 2) A geometrically accurate 3D calculation yields the cell process in Fig. 2 f (arrow 3) and the axon in Fig. 2 f (marked with asterisk). A similar cell process can be found starting from the same cell in Fig. 2 d (lower half): Here, the cell process (red) has apparently penetrated the intact yelinscheide (white). For comparison, Fig. 2 e shows an intact myelin sheath.

5

In summary, CD16 is necessary to control the abnormal homing aberrant CD8 cells into the pyramidal tract, and when this homing process is complete, the cells degrade (lose) the CD16 complex, then migrate as CD8 + CD16 cells between the cells myelinated neurons penetrate the myelin sheaths and axotomize neurons, a process that is apparently primarily autonomous, as the cells are never observed in the context of ZeFdebris, which is a sign of primary neuronal degradation. The cells therefore play a primary pathogenetic role. This interpretation is supported by the fact that downregulation of CD16 leads to a halt of disease progression: when CD16 is no longer available as in Fig. 1 f, the endothelial homing process can no longer be performed. Part of the (sporadic) ALS can therefore be described as a disease of the immune cell toponoma, which generates the ALS-specific pyramidal tract lesions (axotomy) by means of a highly selective "homing code." Many genetic findings and protein aggregations that have been described in ALS are also have been described in experimental axotomy, so that the immunocelltoponomically-mediated axotomy explains these findings.

Koepressionsmuster

The invasive cells according to FIG. 1c further coexpress the following molecules:

Bax, Bcl2, CD1 1 b, CD138, CD16A, CD29, CD2, CD45A, CD49d, CD54, CD56, CD57, CD58, CD62L, CD8, HLADR, immunoglobulin G. MHCII, MHCI, NeuN, SIRT1, RAC1, BMX, GAK, JNK2, MAPKK6, OTUB2, PRKAR2A, SMAD2. S AD4 and STAP2.

NeuN (Fox-3, Rbfox3, or Hexaribonucleotide Binding Protein-3) is a neuronal, nuclear antigen that is normally used as a biomarker for neurons.

Nuclear coexpression of NeuN and CD49d, as well as cytoplasmic expression of immunoglobulin G (IgG) along with the CD16 bearing vesicles of Figure 1df) and cell surface expression of CD8 and CD3 indicate that this cell has abnormal differentiation status: a cell with Merkma - a T cell (CD3, CD8), a neuronal cell (NeuN in the nucleus), a monocyte (CD16) and a B lymphocyte (IgG).

Proteins partially colocalized with specific CD8 and / or CD16 in ALS-indicative cells, in some cases with specific individuality or frequency, are given in Table 1. All of these proteins thus represent therapeutic target structures (targets) individually or in combination with CD8 and / or CD16, since switching off these proteins leads to a collapse of the intracellular information flow and thus to a loss of function of the aberrant cells. Table 1

The cell of Figure 2c, f), which, as stated above, expresses a CD16 negative phenotype upon invasion of the cell of Figure 1, remains positive for NeuN.

Significance: The invasive cell described above can be detected as an ALS-specific cell with the abovementioned features in the blood or in tissue samples by means of spatial tomography analysis. Since their invasion behavior into the pyramidal tract is known as set forth above, and since it is further known that the cell transforms to a NeuN positive T cell by phenotype switching (CD16-), axotomizing the neurons leaves the detection of cells of the CD16 / CD8 phenotype from Fig. 1 c close to the process of axotomy. It follows that cell invasion of the cell of Figure 1c must be therapeutically prevented by molecular blocking or lysis to prevent axotomy and stop the progression of ALS. The described two-step process (homing aberrant "NeuN" positive blood cells into the pyramidal tract, their transformation into "NeuN" positive T cells, which injure the surfaces of nerve cells) is a violation of basic rules of cooperation of cells of different cotyledons: In intact systems, cells from different cotyledons follow the rule of non-injury of the mutual cell surfaces. In the case of ALS, the transformed T-cell-like cells (cotyledon mesenchyme) injure the surfaces of neurons (cotylus ectoderm), resulting in a physical transcellular crosslinking of the two cotyledonous functions with the consequence of the disruption of the nerve tracts to the voluntary musculature. The fact of expression of "NeuN" in the cell nucleus of the acting cells (immune tents in the circulation and after invasion into the nervous system of the pyramidal tract) means that these cells follow a program of aberrant neuronal stem cells, since NeuN is only expressed in neurons under physiological conditions. Thus, NeuN can be used as a marker for the presence of ALS-causing cells, that is for the diagnosis of ALS.

In order to block the molecular "program" of these aberrant cells, that is, as a therapy for treating a patient suffering from ALS, various extracorporeal methods are available to extracorporeally remove the pathogenic cells expressing the lead protein CD16.

For this purpose, the blood of the patient can be subjected to a therapeutic apheresis in which the pathogenic cells are removed from the blood with the aid of preferably immobilized antibodies and / or ligands which bind CD16. Alternatively or additionally, it is also possible to use immobilized antibodies and / or ligands which bind CDS and / or RAC1 and / or STAP2 and / or SMAD2. It is also possible to use a plurality of different antibodies and / or ligands, which are arranged together and / or viewed in the flow direction of the blood, one after the other, resulting in a multistage depletion of the pathogenic constituents of the blood. Alternatively or additionally, it is possible to fractionate the blood prior to extracorporeal removal of the pathogenic cells, for example by centrifugation. Alternatively or additionally, it is possible to treat the blood by photopheresis, the optionally fractionated blood being irradiated for a predetermined time with light of a specific wavelength (s) to inactivate the pathogenic cells, a therapeutic depletion of "ALS cells" means of cells carrying the ALS-specific CMP in the blood of patients suffering from ALS was achieved as follows:

Initially, blood samples from several patients suspected of having amyotrophic lateral sclerosis (ALS) were examined using the imaging cycler technology (ELK / ICM method) with a combinatorial molecular resolution of up to 4.5x1E ⁴⁸¹ . For this purpose, a Toponome Imaging System (TIS) from ToposNomos Ltd, (Munich, Germany) was used. Patients in whom so-called "ALS cells", ie cells with ALS-typical CMPs were detected, were classified as having ALS The patients identified were then subjected to a photopheresis treatment Photopheresis makes it possible to carry out a targeted depletion procedure, so that the number and concentration of "ALS cells" in the blood of the patients considerably, that is to below the detection limit can be reduced. It has been found that the patients felt subjectively better when in a photopheresis cycle more than 1 liter of blood volume, i.e. at least 2 and preferably up to 5 liters of blood volume, was passed through the device for treatment. As demonstrated by the initial examination and subsequent follow-up using Imaging Cycler technology, the cell count of the "ALS cells" in the blood correlates very well with the course of the disease, for example, in one patient with approximately 50 million "ALS cells". per liter of blood less than half as fast as in a patient with about 140 million "ALS cells" per liter of But this as well as the above-discussed finding that exactly these "ALS cells" immigrate after conversion of the cell surface in the pyramidal tract (pathological homing) where they axotomize the axons in the pyramidal tract are further proof that it is the "ALS cells" detected in the blood are the crucial pathogenetically relevant cells and thus a central target of the ALS treatment.

The therapeutic depletion of these "ALS cells" in the blood has always been very reliable with photophore in all patients treated so far.After only a few cycles of photopheresis, typically about 2 to 6 cycles, practically no "ALS cells" could be detected in the blood. Special effects are observed in initial stages of ALS disease. Thus, the disease in the treated patients, according to previous observations for about 5 months or longer no longer progressive, the electrophysiological parameters {EMG etc) show no further progression and the treated patients feel much better. In particular, no or hardly any active EMG activities can be detected in the treated patients. As some patients have shown, "ALS cells" can recur in breaks between photopheresis cycles in the blood, and the recurrent "ALS cells" can be fully depleted again by photopheresis. According to previous knowledge, this procedure can be repeated as often as required, so that at least a progression of ALS disease can be stopped or significantly slowed down. In addition, it was observed that the pathogenic ALS-specific combinatorial CMP motifs of the "ALS cells" decreased at least by a factor of 100 after their return in the blood.The number of disease-specific molecular combinatorics at the cell surface (the so-called combinatorial molecular phenotypes, CMPs, with code (= 1) and anticode (= 0) (Schubert W. et al., Nat. Biotechnol 2006)) of the individual "ALS cells". This effect is stable over the period studied so far. Since cell surface CMPs on the surface of these pathological "ALS cells ^" in the postcapillary, the site of invasion, "slow down" the rolling phenomenon to the stop, the photopheresis-induced decrease in the number of these CMPs will worsen the pathological homing after rolling and thus stop or at least greatly slow down the process of kanking at the pathogenesis site of the invasion.

Further possibilities for extracorporeal removal of the pathogenic cells by means of apheresis methods include: Use of molecules (antibodies / ligands) that bind and / or block / inhibit IgG (1/3) or block / inhibit the physiological mechanism of IgG-mediated activation of CD16. An example of this is antibodies to IgG and (optionally recombinant) streptococcal protein G; Blocking (eg by crosslinking / cross-linking) of the ALS-specific cluster CD16a + CD8 + STTP1, for example by means of at least one bi-, trioder multispecific antibody (recombinant, human, de novo, etc.), which is preferably immobilized on a Matrix bound exists. In this case, STTP1 denotes the optionally patient-specific signal protein (signal transduction protein 1, z, B. kinase), which mediates the signal cascade emanating from the nucleus and coupled together with the module "CD 16a and CD8" on the surface of the cell or with CD 16a and CD8 and colocalized STTP1 can be individually determined or controlled with the help of the known MELK and / or ICM technique (multi-epitope ligand cartography or imaging cycler microscopy) for the respective patient. STTP1 may be, for example, one or more proteins from the group RAC1, STAP2 and / or SMAD2 and / or another protein from the signal transduction chain of ALS cells A corresponding antibody, for example a trispecific anti-CD16a-CD8-STTP1 antibody , can then also be prepared by known manufacturing methods and used for the therapy of the patient in question, since such a trispecific Antik a body has an extremely high selectivity for abnormal, ALS-specific cells, such treatment is particularly low side effects; Use of molecules which bring about the proteolytic separation of the extracellular CD16 domains (ectodomains) (so-called shedding or ecto-domain shedding). An example of this is the use of preferably immobilized Adam17 (metallopeptidase domain 17), which is also called TACE (tumor necrosis factor-a-converting enzyme). Adam 17 is a 70 kDa enzyme belonging to the ADAM protein family of disintegrins and metalloproteases. Adam17 or the respective CD16-Shedding-capable molecule may optionally be coupled to a mono-, bi-, tri- or multispecific antibody (see point 2) to advantageously increase its specificity; 4. Use of at least one mono-, bi-, tri- or multi-specific anti-CD16 antibody (recombinant, human, de novo, etc.), which is preferably immobilized on a matrix material;

5. Use of at least one mono-, bi-, tri- or multi-specific anti-NeuN antibody (recombinant, human, de novo, etc.), which is preferably immobilized on a matrix material;

6. Use of streptamers and / or magnetic beads for the removal of pathogenic CD16 cells.

It can be provided that the pathogenic cells are not or not completely removed from the blood sample, but are inactivated extracorporeal so that they lose their biological functionality. The abovementioned compounds (medicaments) can in principle be used individually or in any combination as part of an apheresis process. The effectiveness and extent of the removal should be checked through regular monitoring and adjusted if necessary. The control can be carried out, for example, by means of the diagnostic method described above. Upon successful removal, significantly fewer or, preferably, no abnormal ALS-specific cells (see above) can usually be detected in the cell or blood sample of the subject patient.

In general, this results in numerous ALS-specific supraindividual and / or individual thenutic options. Superindividual therapy may be directed to the ALS-specific surface proteins (CDS) or surface protein clusters, which are cross-linked, inhibited or bound to a matrix. Additionally, central signaling chain molecules of ALS cells may be cross-linked, inhibited, or otherwise blocked. For example, the cell surface CD8 / CD16A CD45RA may optionally be extracorporeally blocked together with the signal-chain molecule RAC1, since these are exclusively associated directly in ALS cells. Such a therapy can be carried out, for example, with the aid of bi-, tri- and / or tetraspecific antibodies or by means of corresponding variable single-chain fragments (scFv) or multivalent antibody fragments (scFv multimers), ie by so-called diabodies, triabodies or tetrabodies.

Individual targets that may be therapeutically addressed as an alternative or in addition to the ALS-specific cluster described above include individual clusters of CD16 / CD8 with the molecules STAP2 and / or SMAD2, and optionally combinations of the molecules designated in Table 1 together with the CD8 / CD16 clusters or as isolated clusters without CD8 / CD16. These clusters can also be switched off with the help of optionally multispecific antibodies and / or antibody fragments.

Fig. 3 shows a schematic representation of the topological hierarchy of proteins within a toponome. The proteins are symbolized by the symbols star, square, triangle and pentagon. It can be seen that the star-symbolized protein occurs in all three CMPs 1 -3, so that this protein is a lead protein (L, 1, True). In contrast, the protein denoted square does not occur in any of the CMPs 1-3, so that this protein is an anti-colocalized protein (A-absent, 0, False). The other proteins (triangle, pentagon) are variably associated with the lead protein and thus represent so-called wildcard proteins (W, ^* ).

Figure 4 shows a topographic map showing the location of the 30 most common of more than 2,000 different CMPs in prostate cancer. The location of these CMPs in the tissue is shown as an overlay with a CD138 fluorescence signal. Figure 4 illustrates the position of these CMPs by their different colors (or gray levels, respectively) aligned with the CD138 fluorescence signal. Relevant CMPs are shown in Table 2 below and have CD26 and CD29 as lead proteins (= L), while in all of these CMPs CD3 / CD4 / CD8 / CD19 / CD20 / CD44 / CD80 are absent (anti-colocalized, A, 0 ), and CD10 / CD13 / CD38 / CD49d / CD54 / CD58 / CD138 variable associated (W ^*), As mentioned, this results in the symbol code (L, A, W) for the protein clusters (CMP motif). Thus, CD26 and CD29 could be identified as lead proteins on the cell surfaces of prostate tissue cells of a patient affected by prostate cancer.

Table 2

loötdo

Figure 5 shows a representation of the selective expression of the prostate cancer-specific CMP in subcellular sites of neoplastic prostate acini. Figure 5 illustrates the selective expression of CMP1 in subcellular sites of neoplastic prostate acini. CMP1 is the most abundant CMP within the CMP motif of Table 2. Figure 5a shows an overview in which arrow 1 and arrow 2 indicate apical sites of secretory cells, while arrow 3 indicates projections at the basolateral site of an acinus. Fig. 5bc show details indicated by arrows. One recognizes the ring-shaped arrangement of the CMP1 (brown Color) with the cell surface fluorescence signal from CD133 (white). "BE" means basal epithelial cell layer, (bars: 5 ai 100 μηι, 5 bd: 10 ^μηη )

Accordingly, prostate cancer analogous to ALS can be treated by extracorporeal removal of components and cells which have CMPs comprising at least CD26 and / or CD29 as disease-specific lead proteins. For this purpose, for example, an apheresis or photopheresis method can be used in which the CD28 / CD29 expressing cells are separated and / or inactivated or destroyed in the manner described above.

Fig. 6 shows an illustration of the cyclic localization of 25 biomolecules and the subsequent determination of the CMPs by means of the ELK / ICM method, Fig. 8a shows an optical plane of 20 gemisamapped planes in the z-direction and serves to illustrate the fluorescence signals of the in Fig. 7 indicated biomolecules. FIG. 6b illustrates the binarization of the primary signals of FIG. 6a shown in parallel. Figure 6c shows 2D maps of the resulting combinatorial molecular phenotypes (CMPs) calculated for all data points from the binary data set (Figure 6b). Fig. 6d shows two exemplary CMPs from the data points (Fig. 6b).

7 shows topographical images of the tissue organization of a cutaneous lymphoma (bars: in a-d, j: 10 μΐτι, in f, g, k: 1 μιτι). Fig. 7a shows a 3D co-mapping of 3213 CMPs (from 7161 CMPs) within a range of cryosection of a tissue sample of a patient affected by a cutaneous lipomhoma (mycosis fungoides) according to Fig. 7b (boxed area). Different CMPs are coded with different colors. Figure 7b shows a phase contrast image of the cryogenic skin section in which nuclei for histones are stained blue while the basal diamina for CD49f is stained white. Figure 7c shows an enlargement of the boxed area (Figure 7b) and is aligned identically to Figure 7d, showing the lead proteins of the CMPs of Figure 7a. One recognizes an elongated multicellular

Complex of five cell types (cells 1 -5 in a, d, c). An elongated cell projection (arrow 1 in a) of the cells 3, 4, 5 extends from the dermis via the basaliamina (a, BL) into the epidermis, where it is close to CD8 + / CD3 + T cells (cell 2 in a, c, d, brown Color in d and adjacent keratinocytes (cell 1 in a, c, d)). Fig. 7e shows details of a suprabasal part of this structure from different angles of view (e, f, g, h). A cytokeratin-containing cell projection (CK) extends away from the keratinocyte (e, arrow) and projects through the CD8 + / CD3 + T cell surface (compare h, which shows CK in green, compare g, which shows the same without CK). Figure 7k shows a transverse virtual anatomical section through the CD8 + / CD3 + T cell (cell 2 in j). The projection of the keratinocyte penetrates into the T cell (arrow). Figure 7i shows a list of the co-cartier molecules. Fig. 71 specifies the CMP motif characteristic of cutaneous lymphoma:

L: HLA DQ

A: CD2, CD10, CD18, CD54, CD57, CD58, CD62L, CD80

W: CD3, CD4, CD7, CD8, CD13 _» CD26, CD29, CD36, CD44, CD45, CD49f, CD56, CD71, HLA-DR, Cytok., Histone.

Accordingly, cutaneous lymphoma may be treated analogously to ALS by extracorporeal removal of components and cells having CMPs comprising at least HLA-DQ as a disease-specific lead protein. For this purpose, for example, an apheresis or photopheresis method can be used in which the HLA-DQ expressing cells are separated and / or inactivated or destroyed in the manner described above.

The method may therefore, in principle, comprise a combination of toponomous technology with methods of isolating cells from body fluids and / or tissues. By using toponom technology, cells can pass through

Coagulation of thousands of multiprotein complexes are classified extremely accurately. In this way the disease-specific cells or CMPs are found. The cells in question can then be known per se

Specific methods are isolated and used for various purposes, such. As a cloning, extracorporeal expansion, re-transfer into tissue or blood, therapeutic applications, etc. A particular aspect is the isolation of disease-specific cells from the blood circulation, which migrate as autoimmune cells or aberrant cells of the immune system in organs where they targeted tissue structures to destroy. In the latter case z. B. the isolation of such cells from the blood circulation by a therapeutic apheresis / photopheresis by therapeutic apheresis / photopheresis a very targeted measure aimed at halting disease progression, as the disease-specific cells can be precisely predefined by high-dimensional toponomy mapping so that they can be removed from the circulation in a targeted manner via antibody-based isolation. By consecutive monitoring by toponomic mapping of the blood, the efficiency of the procedure can be controlled. Another advantage is the use of such cells for the development of drugs for the selective blocking / elimination of such cells or, in the case of stem cells, for reinfusion into the organism (e.g., therapeutic organ regeneration or tumor therapy), or for use with the development of diagnostics, or for the treatment of amyotrophic lateral sclerosis (ALS) or other diseases, by removing pathogenic cells from the blood-clotting, to prevent their biological mechanism, in the case of ALS invasion into the motor neuron system and its neurotoxic damage mechanisms.

Fig. 8 shows a schematic representation of the pathomechanism of ALS. A normal or healthy T cell 10 first matures in an extrathymic environment 12 (skin) and is delivered to the bloodstream 14. Due to the lack of Hönning codes, the normal T cell 10 can (and should not) invade the pyramidal system 16, which is responsible for fine motor skills and voluntary motor function in mammals. In contrast, "ALS cells" represent 18 aberrant T cells with a defective homing code that can be characterized by the CMPs described above Accordingly, blood circulating ALS cells 18 can enter the pyramidal system 16 from the blood They axoto- mize neurons, which leads to a degeneration of the motor nervous system and thus to the ALS-typical clinical picture with progressive paralysis of the patient.The aim of an ALS treatment must therefore be the formation of ALS cells 18 and / or the invasion of ALS cells For example, ALS cells 18 can be deactivated by blocking the lead proteins and / or by inducing apoptosis (deactivated ALS cells 18 '), thereby losing their biological functionality and not can migrate longer in the pyramidal system 16. 9 shows a schematic representation of a therapy of ALS, aberrant ALS cells 18 mature as already mentioned in an extrathymic environment 12 (skin) and are delivered into the bloodstream 14. By a therapeutic treatment 20 of the blood 14, for example by photopheresis and / or by deactivation of the ALS-lead proteins CD8 and CD16 by cross-linking with bi- or multispecific antibodies, the ALS cells 18 circulating in the blood stream 14 become apoptotic (deactivated ALS cells). Line 18 ') and can no longer migrate into the pyramidal system 16. Apoptotic bodies 22 of the deactivated ALS cells 18 'are formed. These apoptotic bodies 22 are taken up by neighboring cells 24 (phagocytosis) and degraded. The apoptotic bodies 22 are also presented to the immune system. Based on the current state of knowledge, it can be assumed with high probability that this mechanism can also induce an endogenous immune response against the aberrant ALS lines 18, so that the extrathymic maturation of the aberrant ALS cells 18 can be at least partially suppressed and the number of replicated ALS lines 18 decreases.

The parameter values given in the documents for defining process and measurement conditions for the characterization of specific properties of the subject invention are also within the scope of deviations - for example due to measurement errors, system errors, weight errors, DIN tolerances and the like - as being within the scope of the invention to see included.

Claims

claims

A method for extracorporeal removal of pathogenic and / or supernumerary components from a cell sample of a human or animal patient suffering from a disease, comprising the steps of: a) determining at least one protein cluster (CMP) characteristic of the disease of the patient;

b) providing the patient's cell sample; and

c) extracorporeal removal of components comprising the at least one detected CMP from the cell sample.

Method according to claim 1,

characterized in that

a body fluid, in particular blood and / or blood plasma, and / or a tissue sample of the patient is provided as a cell sample.

Method according to claim 1 or 2,

characterized in that

the at least one CMP is determined by means of a database and / or by means of a multi-epitope ligand cartography (MELK) or ICM method (multi-epitope ligand cartography / imaging cycler microscopy) and / or by means of a MELK robot system, and / or that the extracorporeal removal of the components is monitored and / or controlled by means of a MELK or ICM method and / or by means of a MELK robot system.

Method according to one of claims 1 to 3,

characterized in that

the disease is a tumor disease and / or an inflammatory disease.

5. The method according to any one of claims 1 to 4,

characterized in that the extracorporeal removal is carried out by means of an apheresis method and / or by means of an apheresis device.

Method according to claim 5,

characterized in that

as an apheresis method, an unselective apheresis and / or a selective apheresis and / or a whole blood apheresis and / or a photopheresis is performed and / or that the apheresis device for performing at least one apheresis method from the group unselective apheresis, selective apheresis, whole blood apheresis and photopheresis is formed.

Method according to one of claims 1 to 6,

characterized in that

for extracorporeal removal at least one streptamer and / or at least one antibody and / or at least one ligand, in particular a variable single chain fragment (scFv) and / or a multivalent antibody fragment (scFv multimer) is used.

Method according to claim 7,

characterized in that

the at least one streptamer and / or the at least one antibody and / or the at least one ligand specifically binds and / or inactivates a lead protein characteristic of the disease.

Method according to one of claims 1 to 8,

characterized in that

the disease is amyotrophic lateral sclerosis (ALS) and / or that the cell sample is a blood sample of the patient and / or that the at least one CMP is one or more of CD16, CD8, NeuN, Bax, Bcl2, CD11b, CD138, CD16A, CD29 , CD2, CD45RA, CD49d, CD54, CD56, CD57, CD58, CD62L, CD3, HLADR, immunoglobulin G, MHCII, MHCI, SIRT1, RAC1, BMX, GAK, JNK2, MAPKK6, OTUB2, PRKAR2A, SMAD2, SMAD4 and STAP2 and / or that the constituents are produced by means of at least one antibody and / or ligand, which is at least one of the group CD16, CD8, NeuN, Bax, Bcl2, CD11b, CD138, CD16A, CD29, CD2, CD45RA, CD49d, CD54, CD56, CD57, CD58, CD62L, CD3, HLADR, immunoglobulin G, MHCII, MHCI, SIRT1, RAC1, BMX, GAK, JNK2, MAPKK6, OTUB2,

PRKAR2A, SMAD2, SMAD4 and STAP2 bind, and / or are extracorporeally removed by means of a CD16 ectodomain shedding molecule.

10. The method according to claim 9,

characterized in that

for extracorporeal removal, an antibody and / or ligand is used which binds two or more of the group CD16, CD8, RAC1, STAP2 and SMAD2.

1 1. A method according to any one of claims 1 to 8,

characterized in that

the disease is prostate cancer and / or that the cell sample is prostate tissue and / or that the at least one CMP comprises one or more of the group CD26 and CD29 and / or that the components are at least one antibody and / or ligand, which is at least one of the Group CD26 and CD29 binds to be removed extracorporeally.

12. The method according to claim 1 1,

characterized in that

the at least one CMP also comprises CD44 and / or CD54 and / or CD138 and / or in which at least one of CD3, CD4, CD8, CD10, CD13, CD19, CD20, CD38, CD49d, CD58 and CD80 is missing.

13. The method of claim 1 1 or 12,

characterized in that for extracorporeal removal an antibody and / or ligand is used which binds at least one or more of the group CD26, CD29, CD44, CD54 and CD138.

14. The method according to any one of claims 1 to 8,

characterized in that

the disease is a cutaneous lymphoma and / or that the cell sample is a Skin sample is and / or that the at least one CMP comprises one or more of the group HLA-DQ, CD2, CD3, CD4, CD7, CD8, CD10, CD13, CD18, CD18, CD26, CD29, CD36, CD44, CD45, CD49f, CD54, CD56, CD57, CD58, CD62L, CD71, CD80 and HLA-DR comprises and / or that the components by means of at least one antibody and / or ligand, which at least one of the group HLA-DQ, CD2, CD3, CD4, CD7 , CD8, CD10, CD13, CD18, CD18, CD26, CD29, CD36, CD44, CD45, CD49f, CD54, CD56, CD57, CD58, CD62L, CD71, CD80 and HLA-DR, can be extracorporeally removed.

Method according to one of claims 1 to 14,

characterized in that

after the partial or complete extracorporeal removal of the components having the at least one detected CMP, the cell sample is returned to the patient.

16. Device which is designed for carrying out a method according to one of claims 1 to 15. 17. The device according to claim 16, which for carrying out at least one

Apheresis method from the group unselective apheresis, selective Aphere- se, whole blood apheresis and photopheresis is formed.

18. Streptamer and / or antibody and / or ligand for use in a method according to any one of claims 1 to 15.