7 Diarrhoea and constipation. K. Ewe. x. Faivle, SA, Cunningham, JA, and Komorowski, P. Über die Häufigkeit von Leberschädigungen unter INH-Therapie.

Government support under Grant Number K08 KHLA and P01 CA, awarded by National Institutes of Health. The United States Government has certain rights in the invention. Specifically, IL can be used to increase Intestinal Stem Cell ISC recovery and for enhancing immune reconstitution following allogeneic hematopoietic transplantation.

In Komorowski über Psoriasis preferred embodiments, the present invention provides methods of using therapeutic IL, including a dimeric form of IL, in therapeutic compositions for treating graft vs. Current strategies to reduce clinical GVHD have the undesired effect of limiting both post-transplant immune function and therapeutic beneficial graft vs. Of particular concern is the maintenance and regeneration of intestinal epithelial tissues during graft vs.

IL increases recovery of ISCs from immune-mediated pathology by accelerating regeneration of the ISC pool. This resulted in augmented recovery of mature epithelium without augmenting growth factor production by the stem cell niche.

The method does not require immunosupression to achieve its therapeutic effect. In one embodiment, IL is administered to the subject beginning from 1 day to 6 months following transplant; Komorowski über Psoriasis one embodiment, IL is administered to the subject beginning from Komorowski über Psoriasis week to 4 months following transplant. Specific polypeptide sequences are described in U.

The IL polypeptides used herein may be isolated from a more info of sources, such as from Komorowski über Psoriasis tissue types or from another source, or prepared by recombinant or synthetic methods.

For example, descriptions of the preparation of, purification of, derivation of, formation of antibodies to or against, administration of, compositions containing, treatment of article source disease with, etc. Additionally, the IL Komorowski über Psoriasis use in the present inventions may be a product of a recombinant method wherein the IL encoding DNA is administered Komorowski über Psoriasis a subject, for example, such as lactobacilli expressing IL The term "IL polypeptide" also includes variants of the IL- 22 polypeptides.

The IL of the present invention may Komorowski über Psoriasis be modified in a way to form a chimeric molecule comprising IL fused to another, heterologous polypeptide or amino acid sequence. The term IL as used herein, includes both a monomer and Komorowski über Psoriasis dimer form of IL However unless otherwise specified herein rIL was used in these methods. IL is also known as interleukin related T cell-derived inducible factor IL-TIF.

In some embodiments, an IL dimer contains two duplicate IL molecules, in other embodiments, an IL dimer is made up of different IL proteins. Further examples of IL- 22 dimers that may find use in the present inventions are described in United States Patent Applicationherein incorporated by reference in its entirety.

One suitable IL dimer is a recombinant IL dimerized Komorowski über Psoriasis containing human interleukin 22 IL and produced in continue reading Chinese Hamster Ovary CHO cells in serum-free culture produced by Generon Shanghai Corporation Ltd. IL dimers are described, for example, in United States Patent Applicationincluding sequence information, herein incorporated by reference in its entirety.

IL dimer forming polypeptides used herein may be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods. In Komorowski über Psoriasis embodiments, an IL Komorowski über Psoriasis comprises a carrier protein, including but not limited to an Fc fragment of human IgG 1, 2, 3, 4or human albumin. IL can be localized at the C-terminal Komorowski über Psoriasis N-terminal Komorowski über Psoriasis the carrier protein.

In some embodiments, each monomeric unit of the IL dimer comprises an IL domain linked to a dimerization domain via an optional linker sequence, such as a linker sequence that is about 5 to about 50 amino acids.

In some embodiments, the dimerization domain comprises at least two cysteines capable of forming intermolecular disulfide bonds. In some embodiments, the dimerization domain comprises at least a portion of the Fc fragment. In some embodiments, the Fc fragment comprises CH2 and Komorowski über Psoriasis domains. In some embodiments, the IL dimers comprise two monomeric units as described Komorowski über Psoriasis United States Patent Applicationincorporated specifically herein by reference.

In some embodiments, the IL dimer is administered intravenously. As examples, Komorowski über Psoriasis who have had a blood or marrow transplant from someone else are at risk Komorowski über Psoriasis having acute GVHD. Even donors who are HLA-matched with the recipient can cause GVHD because the donor cells can potentially also make an immune response against minor antigen differences in the recipient.

Acute graft-versus-host disease GVHD is specifically a disorder caused by donor immune cells in patients who have Komorowski über Psoriasis an allogeneic marrow or blood cell transplantation. The most commonly affected tissues are skin intestine and liver. In severe cases, GVHD can cause blistering in the skin or excessive diarrhea and wasting.

Also, inflammation caused by donor immune cells in the liver can cause obstruction that causes jaundice. Other tissues such as lung and thymus may also become affected. The diagnosis check this out usually confirmed by looking at a small piece of skin, liver, stomach or intestine with a microscope for observation of specific inflammatory characteristics.

In severe cases, the liver does not function properly to eliminate waste products from the body. Acute GVHD usually begins during the first 3 months after the transplant.

In some cases, it can persist, come back or begin more than 3 months after the transplant. Other immunosuppressive medications are used if treatment with prednisone is not successful, even though a large proportion of patients is refractory to immunosuppressive medication and die. Older patients, patients who received a peripheral blood instead of bone marrow transplant, and patients who had a mismatched or unrelated donor have a greater risk of chronic GVHD.

Chronic GVHD usually begins later after transplant and lasts longer than acute GVHD. Patients with Chronic GVHD may present with a wide variety of symptoms. Unlike acute GVHD, chronic GVHD can cause damage in the glands that produce tears in the eyes and saliva in the mouth resulting in dry eyes or a dry mouth.

Patients may have mouth ulcers causing pain while eating, skin rashes, or liver inflammation. Chronic GVHD can also cause many other problems. One such problem is formation of scar tissue in the skin cutaneous sclerosis and joints.

Another such problem is chronic damage to air passages in the lungs bronchiolitis obliterans syndrome. Prednisone or other similar antiinflammatory or immunosuppressive medications are used to treat chronic graft-versus-host disease. Other immunosuppressive medications can be used if treatment with prednisone is not successful. Just as in acute GVHD a large proportion of patients is not cured from chronic GVHD.

In severe cases, GI- GVHD can cause pain in the abdomen and bleeding in the stomach or intestines. In other words, the patient receives bone marrow or blood stem cells from a tissue -matched or a close matched donor, i.

Identical twin allogeneic transplants are called Komorowski über Psoriasis transplants. Such allogeneic HSCT is done in patients having certain cancers of the blood or bone marrow, such as multiple myeloma or leukemia, congenital immunodeficiencies and bone marrow failures or other hematologic disease. Infection and graft-versus-host disease is a major complication of HSCT.

It is not intended to be limited to live or functioning cells. The contacting may be accomplished using any suitable method. For example, in one embodiment, contacting is by adding the compound to a tube of cells.

Contacting may also be accomplished by adding the compound to cells in a microtiter plate. Contacting may also be accomplished by adding the compound to a culture of the cells or to an organoid culture. It is not meant to limit how the compound contacts the cells. In one embodiment, contacting may be accomplished by administration of the compound, such as an IL molecule composition to an animal in vivo. It is not intended that the term be limited to any particular cell culture medium.

For example, it is intended that the definition encompass outgrowth as well as maintenance media. Indeed, it is intended that the term encompass see more culture medium suitable for the growth of the cell cultures of interest.

In one sense it can refer to an animal cell or tissue including a human cell or tissue. In another sense, it is meant to include a specimen or culture obtained from any source, in particular as a biological sample. Biological samples may be obtained from animals including humans and encompass fluids, solids, gases, tissues, cells, blood bone marrow and bones.

Thus, primary isolation of cells, such as cells isolated directly from mice or Komorowski über Psoriasis used for flow cytometry analysis, involves such processes as removing tissue from a subject, such as a bone marrow sample, or intestinal sample, etc. Primary isolation may be accomplished using solid or semi-solid agar media, or in liquid. For example, where contacting results in at least a "portion" of said Komorowski über Psoriasis population, it should be clear that portion is with reference to a population.

The fragments may range in size from read more amino acid residues to the entire amino sequence minus one amino Komorowski über Psoriasis. Increased function may increase a cell type, such as when "Paneth cell production is Komorowski über Psoriasis or when "lymphoid cell production is increased", etc.

Thus, examples of increased mature functional lymphoid cells from bone marrow are increased enhanced numbers of at least one lymphoid cell type, such Komorowski über Psoriasis immature B cells, a pre-B cell, etc. Further, examples of contemplated enhanced or enhancing immune function may also refer to enhancing immunocompetence, such as methods wherein immunocompromised patients having reduced immune function, such as reduced T-cell antigen receptor repertoires, reduced cytokine secretion, reduced proliferative responses to antigen, and the like, Komorowski über Psoriasis treated with compositions comprising IL resulting in increased immune function.

Increased immune function may be measured as an increase in any one of T-cell antigen receptor repertoires, cytokine secretion, proliferative responses to antigen, ability to respond to infections, and Komorowski über Psoriasis like.

Another example of reduced function may be a lowered production of a cytokine, i. As other examples, a reduced function is thymic epithelial cells producing a lower number of mature functional thymocytes, a decreased function is bone marrow cells producing a reduced number of mature functional lymphoid Komorowski über Psoriasis, bone marrow derived stem cells have decreased numbers of lymphoid cells.

In other words, Komorowski über Psoriasis reduced function may also be an altered function, for one example, altered bone marrow immune cell generation may show that bone marrow derived stem cells have or produce decreased lymphoid cells. Having a reduced function is not meant to be a static result. In some Komorowski über Psoriasis, contacting or administering an IL composition of the present inventions may alter a function, such as when IL induces cells having reduced function to display increased function, see Examples.

Readouts for in vitro assays of organoid cultures could include for examples, flow cytometry measurements of intestinal stem cell progeny, such as basal crypt cells. Readouts for in vitro assays of immune function could include for Komorowski über Psoriasis, T cell functional assays, including cytokine production, T cell subtypes, granulocytes, stromal cells, proliferation, extent of apoptosis, etc.

Typically, the terms "subject" and "patient" are used interchangeably herein, particularly in reference to a "human subject. Examples of immunocompromised subjects include subjects that have any of the following conditions, chemotherapy, exposure to radiation, deliberate irradiation, Komorowski über Psoriasis immunodeficiency virus infections, transplantation, etc.

Thus "treatment" refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down lessen the targeted pathologic condition or disorder. Subjects in need of treatment include those already with the disorder as well as those prone to have the disorder or those in whom the disorder is to be prevented. The present invention is directed towards treating patients with medical Komorowski über Psoriasis relating to a loss of immunocompetence from a treatment related Komorowski über Psoriasis a disease such Komorowski über Psoriasis irradiation, chemotherapy, immunosuppression, etc.

Accordingly, Komorowski über Psoriasis treatment of the invention would involve preventing, inhibiting or relieving any medical condition where Komorowski über Psoriasis desired level of immunocompetence would be achieved by the use of an IL composition of the present inventions.

In certain embodiments, treatment refers to exposing a subject to a Komorowski über Psoriasis directed towards treating a disease, such as irradiation, chemotherapy, and the like. Acceptable routes of administration to the human body can be through the eyes ophthalmicmouth oralskin transdermalnose nasallungs inhalantmucosal e. Administration "in combination with" one or more further therapeutic agents includes simultaneous concurrent Komorowski über Psoriasis consecutive administration in any order.

An "effective amount" may be determined empirically and in a routine manner, in relation to the stated purpose. Examples of effective amounts On using the pharmaceutical composition, a safe and effective amount of the IL dimer of the present invention is administrated to a mammal e. As one example, for a human of 60kg, the administration dosage is usually 0. The compositions also may include stabilizers source preservatives.

Examples of carriers, stabilizers, Komorowski über Psoriasis adjuvants are described in the art See e. Examples of forms of IL Komorowski über Psoriasis the present invention used for administration comprise ointment, powder, patch, sprayer, and inhalant.

It is intended to include homologues and synthetic versions. Examples include IL, IL, IL, the IL family, IL-7, the interferon IFN family, CC chemokines, CXC chemokines, and the like. As used herein the term "recombinant protein" or "recombinant polypeptide" refers to a protein molecule that is expressed from a recombinant DNA Komorowski über Psoriasis e. The order of these deoxyribonucleotides determines the order of amino acids along the polypeptide protein chain.

The DNA sequence thus codes for the amino acid sequence. A genomic form of a genetic clone contains the coding region interrupted with non-coding sequences termed "introns" or "intervening regions" or "intervening sequences. Introns are removed or "spliced out" from the nuclear or primary transcript; introns therefore are absent in the messenger RNA mRNA transcript. The mRNA functions during translation to specify the sequence or order of amino acids in a nascent polypeptide.

These terms encompass the case where there has been amplification of one or more segments of one or more target sequences. This Komorowski über Psoriasis for amplifying the target sequence consists of introducing a large excess of two oligonucleotide primers to Komorowski über Psoriasis DNA mixture containing the desired target sequence, followed by a precise sequence of thermal cycling in the presence of a DNA polymerase.

The two primers are complementary to their respective strands of the double stranded target Komorowski über Psoriasis. To effect amplification, the mixture is denatured and click primers then annealed to their complementary sequences within the target molecule.

Following annealing, the primers are extended with a polymerase so as to form a new pair of complementary strands. The steps of denaturation, primer annealing, and polymerase extension can be repeated many times i. The length of the amplified segment of the desired target sequence is determined by the relative positions of the primers with respect to each other, and therefore, this length is a controllable parameter.

By virtue of the repeating aspect of the process, the method is referred to as the "polymerase chain reaction" hereinafter "PCR". Because the desired amplified segments of the target sequence become the predominant sequences in terms of concentration in the mixture, they are said to be "PCR amplified. In particular, the amplified segments created by the PCR process itself are, themselves, efficient templates for subsequent PCR amplifications.

With PCR, it is possible to amplify a single copy of a specific target sequence in genomic DNA to a level detectable by the device and systems of the present invention. Typically, amplification reagents along with other reaction components are placed and contained in a reaction vessel test tube, microwell, etc. The starting mRNA is enzymatically converted to complementary DNA or "cDNA" using a reverse transcriptase enzyme.

The cDNA is then used as a "template" for a "PCR" reaction. It is based on detection of a Komorowski über Psoriasis signal produced proportionally during amplification of a PCR product. The primer is preferably single stranded for maximum efficiency in amplification, but may alternatively be double stranded. If double stranded, the primer is first treated to separate its strands before being used to prepare extension products.

Preferably, the primer is an oligodeoxyribonucleotide. The primer should be sufficiently long to prime the synthesis of extension products in the presence of the inducing agent. The exact lengths of the primers will depend on many factors, including temperature, source of primer and the use of the method. In contrast, "background template" is used in reference to nucleic acid other than sample template that may or may not be present in a sample.

Background template is most often inadvertent. It may be the result of carryover, or it may be due to the presence of nucleic acid contaminants sought to be purified away from the sample. For example, nucleic acids from organisms other than those to be detected may be present as main celandine Salbe Schuppenflechte Behandlungsmethode in a Komorowski über Psoriasis sample.

A, KO recipients, minor antigen mismatch. B, KO recipients, MHC mismatch. C, Anti-IL neutralizing antibody, MHC mismatch. D, KO marrow, MHC mismatch. E, KO donor T cells, MHC mismatch. F, Hematopoietic KO recipients, minor antigen mismatch with reduced intensity BMT 3 months after hematopoietic chimeras were generated by reconstituting WT B6 mice with IL KO or WT congenic CD Data representative D or combined from independent experiments.

C, qPCR of lamina propria cells 2 weeks after B E, Serum IL after allo-BMT with or without GVHD. F, ELISA on intestinal homogenates from B6 mice combined from two experiments. A, SI crypt schematic. C, IF for ILR greenPaneth cell lysozyme redand nuclei blue. This staining, and thus the survival of ISC, is still present in the absence of GVHD bottom leftbut staining and stem cells are significantly lost during GVHD bottom right. Combined statistics for CBC counts are also shown upper left. E, Assessment of ISC histologically based on niche location at the crypt bases between Paneth cells in WT or IL KO mice.

FITC-dextran translocation across epithelium into the blood stream indicates loss of barrier integrity. LP- B6 3 weeks post- BMT, rIL administration daily starting day 7 post-BMT. A, Decreased gut GVHD pathology after IL administration.

No observed change was found in skin GVHD after IL administration. Decreased crypt apoptosis after IL administration. Paneth cells were decreased in GVHD and were not increased by IL administration.

F, No observed change was found expression of Paneth cell- derived molecules after IL Komorowski über Psoriasis, Innate antimicrobial genes are increased in GVHD after IL- 22 treatment. All data representative or combined from at least two experiments and at least five mice per group. Organoids grown ex vivo from small intestine crypts. Crypts were isolated from Komorowski über Psoriasis small intestine and cultured in matrigel with R-spondinl, EGF, and noggin.

Sphere formation from crypt after 24 hours in culture. Organoid with new crypt buds after 7 days in culture. IL dimer augmentation of ISC function. IL receptor expression on SI ISCs isolated from Lgr5-GFP reporter mice: High magnification of in vitro organoid demonstrating perimeter tracing for measurement of organoid perimeter and area.

Increased size of SI organoids after culture with IL for one week. Organoid size after culture with murine IL Organoid size after culture with F human IL dimer. Data combined or representative of at least two independent experiments. GVL Komorowski über Psoriasis A20 lymphoma. IL KO donor T cells. Donor T cell IL was not observed to affect GVL against A20 lymphoma. Komorowski über Psoriasis was equivalent in recipients of WT and IL KO T cells after tumor challenge with A B, WT T cells and daily IP PBS vs.

Potential alternative GVL model: Donor T cells mediate GVT against AML generated by lentiviral transduction with MLL-AF9 fusion construct. HCT with MLL- AF9 leukemia. Daily IP administration with rIL protein. Decreased GVHD pathology was observed in recipient small intestine, large intestine, and liver three weeks post-HCT. Recipients treated with rIL protein. Marrow-derived T cells 4 weeks post-transplant with rIL administration.

A, intestinal GVHD pathology score B, intestinal crypt Komorowski über Psoriasis score. C, IL ELISA after lactobacillus culture Komorowski über Psoriasis vitro, left bar is lactos. D, IL FACS of ILsurface-anchored lactobacillus with pink or without blue pH adjustment, and negative control lactobacillus green.

IL administration decreases GVHD pathology. LP into B6, 3 weeks post-BMT. IL administration increases expression of innate antimicrobials. Representative images of early budding are shown in ascending order.

A indicates polarization prior to budding. IL reduces intestinal tissue pathology without substantially altering alloreactive immunity. Data combined or representative of two independent Komorowski über Psoriasis with at least nine mice per group. Data are combined Komorowski über Psoriasis two independent experiments with at least seven mice per group, a, LP- B6 BMT was performed with Lgr5- LacZ reporter mice as recipients.

Recipient mice were monitored for clinical signs of GVHD h and GVHD-related mortality i. F treatment reduces the severity Komorowski über Psoriasis FOX3P deletion. This disrupts crypt homeostasis and negatively affects the intestinal stem cell compartment Komorowski über Psoriasis evidenced by impaired organoids growth from small intestine SI.

SI crypts were harvested eight days Komorowski über Psoriasis DT treatment. F treatment improves overall survival and reduces systemic markers of GVHD in vivo.

A-B experiments were a MHC-matched allogeneic bone marrow transplant model with 10 mice per group in each of the treatment groups, 5 in the bone marrow only Komorowski über Psoriasis. These mice started treatment with F or PBS seven days post bone marrow transplant.

C-D experiments were a MHC-matched allogeneic bone marrow transplant model with 5 mice per group in each of the groups receiving treatment with F or PBS only after systemic GVHD symptoms started to develop four weeks post-BMT. However IL is not currently Komorowski über Psoriasis as a treatment for human GVHD. Komorowski über Psoriasis fact, the inventors found that treatment of GVHD with donor T cells engineered to Komorowski über Psoriasis IL may cause undesirable effects such as increased skin GVHD.

A description of other methods, vectors and host cells for synthesis of an IL for use in practicing the click of the present invention can be found in Gething et al, Nature, Recombinant IL, dimers and fusion proteins thereof are produced using methods known to those of skill in the art.

For more details, see U. Alternatively, DNA encoding IL can be obtained by PCR amplification or synthesis and then joined together to form a DNA sequence encoding an IL- 22 dimer. Cold Spring Harbor Komorowski über Psoriasis Press Suitable mammalian cells include Chinese Hamster Ovary CHOCOS cells; in particular, SVtransformed monkey kidney CV1 cell line COS-7, ATCC CRL ; human embryo kidney cell line Graham et al, J.

Many vectors for protein expression are known in the art. Using these techniques, a nucleic acid sequence encoding IL- 22 is inserted into an appropriate vector, which may further include any of the following: One skilled in the art will be able to select a suitable method depending on the host cell selected.

In general, a pharmaceutical composition of IL comprises 0. Depletion of T cells abrogated GVHD and GVL effects. Thus in IL knock-out mouse and irradiated mouse models, IL was produced by recipient-derived innate lymphoid cells ILCs and contemplated to provide a signal for epithelial recovery following experimental allogeneic hematopoietic cell transplantation allo-HCT.

Komorowski über Psoriasis ILdeficient recipients demonstrated increased GVHD mortality and significantly worse loss of crypt base intestinal stem Komorowski über Psoriasis ISCs during GVHD. Paradoxically, GVHD also led to reduced gastrointestinal GI IL levels in irradiated wild-type WT recipients due to the elimination of radioresistant intestinal cells ILCs. Therefore the inventors were Komorowski über Psoriasis to discover that IL administration after allo-HCT negated reduced the effect of ILC Komorowski über Psoriasis and further reduced GVHD pathology without impairing graft versus leukemia GvL responses.

Loss of ISCs in GVHD may be due to the loss of their niche, or the loss of Paneth cells may be due to the loss of their parental ISCs. The villus epithelium of the SI is thought to renew every days in normal homeostasis, while Paneth cells are more long-lived, lasting for three weeks or even longer. These kinetics may be altered after tissue damage and in the transplant setting specifically.

In addition to lineage tracing, intestinal tissue will be harvested to measure RNA and protein expression of reported ISC genes Lgr5, BMI-1, Hopx, mTert, LriglPaneth cell genes involved with ISC function Wnt3, EGF by quantitative q PCR and western blot.

Crypt function and Paneth cell renewal post-BMT are altered due to the observed Paneth cell loss in GVHD. Loss is due to impaired stem cell function or due to immune -mediated loss of the stem cells will be determined.

Evaluation of kinetics of ISC and Paneth cell loss in GVHD can be done to identify the earliest evidence of damage to ISCs and their niche. Lineage tracing of epithelial cells in Lgr5-LacZ lineage-tracing reporter mice can be conducted as described herein, however in an alio transplant setting.

The expression of ISC and Paneth cell related Alter die behandelte Nagel-Psoriasis Forum the Komorowski über Psoriasis, BMI-1, Hopx, mTert, Lrigl, Wnt3, EGF can be measured again. The earliest direct evidence of damage to the ISC compartment can be established in order to determine if ISC loss or Paneth cell loss occurs prior to the Komorowski über Psoriasis in GVHD.

Additionally, evaluating Komorowski über Psoriasis epithelial renewal after treatment with non-myeloablative and ablative doses of chemotherapy may be done by evaluating Komorowski über Psoriasis damage to the ISC compartment in patients having undergone clinical transplantation. Therefore studies described herein and contemplated experiments are described herein Komorowski über Psoriasis evaluate Paneth Komorowski über Psoriasis in the SI in GVHD.

ISCs are reduced in GVHD through the use of genetic, phenotypic, and histologic markers Figs. Stem cells can be evaluated functionally in GVHD by their ability to source intestinal organoids in vitro. Crypts were isolated Komorowski über Psoriasis the small and LI from intestinal epithelium and cultured in semisolid medium in the presence of stem cell growth factors R-spondinl, EGF, noggin for SI; R-spondinl, EGF, noggin, HGF, and Wnt3a for LI.

As each crypt contains a functional stem cell compartment with ISCs and supportive niche cells, crypts cultured ex vivo grow into organoids with crypt buds that recapitulate the in vivo intestinal organization with crypt-villus structures and central lumens Fig. In addition, Lgr5-GFP reporter mice can be used as transplant recipients so that single ISCs can be isolated by Komorowski über Psoriasis post-BMT and cultured in vitro to evaluate the function of isolated ISCs in Read article, providing a functional read out of the stem cell niche and specifically ISCs in GVHD.

Indeed, the direct interactions occurring between alio T cells and recipient epithelial cells are not known. In order to visualize and efficiently test the mechanisms without limiting the invention to Komorowski über Psoriasis particular mechanism of T cell-mediated damage to the stem cell niche, an in vitro system modeling gut GVHD Komorowski über Psoriasis T cells directly interacting with GI epithelium cultured Komorowski über Psoriasis vivo can be tested.

In preliminary studies, crypts cultured in vitro in the presence of alio T cells have a reduced capacity to grow into organoids. Crypts can be assessed for their capacity to grow into organoids. Organoids can be counted and measured for diameter and area by light microscopy.

High resolution three-dimensional images Komorowski über Psoriasis also be Komorowski über Psoriasis by confocal microscopy. In addition to culturing intact crypts, the above experiments can be repeated by culturing individual ISCs isolated from Lgr5-GFP reporter Komorowski über Psoriasis with alio T Komorowski über Psoriasis and measuring organoid formation as above. This also allows for direct imaging of ISCs in cultured organoids by expression of GFP, and also allows for evaluation of interactions between alio T cells and ISCs to determine if alio T cells can directly eliminate or Komorowski über Psoriasis the function of ISCs.

Additionally, this would provide an added benefit of being able to culture cells from mice of additional backgrounds. Assess ISC and niche cell expression of death receptors before and after BMT. Paneth cells, or the loss of Paneth cells may be due to the loss of their precursor ISCs. Alternatively, there may be comprehensive nonspecific damage to the ISC compartment mediated by alio T cells attacking both the ISCs and niche cells. Expression can be compared at baseline, after TCD BMT, and in mice with GVHD.

We can also compare ISCs and Paneth cells by their expression of MHC class I and II and their expression of molecules that could provide resistance to apoptosis BCl-2, Bcl-6, and c-FLIP.

Function of T cell cytotoxic molecules in damaging the ISC compartment. We can test the ability of T cells deficient for cytotoxic pathways to damage ISCs and their niche in vivo and in vitro. SI and LI crypts can be cultured with alio T cells from mice deficient in secreted or cell surface death receptor ligands FasL, TNFa, IFNy, TRAIL read more well as perforin and evaluate the number and size of organoid development as described herein.

Komorowski über Psoriasis cells with impaired ability to damage continue reading in vitro are then evaluated click at this page vivo by performing alio BMT with deficient T cells and assessing histologically post-BMT for ISC and Paneth cell elimination.

Komorowski über Psoriasis, specific damage to the ISC and niche cell compartment in clinical GVHD is poorly understood.

Therefore, we can evaluate the expression of ISC-related molecules in BMT Komorowski über Psoriasis. Biopsy specimens for research purposes Komorowski über Psoriasis be acquired under MSKCC IRB protocol from patients undergoing clinically indicated GI biopsies related to symptoms post-transplant.

Samples are compared between patients found to have biopsy proven GI GVHD and those without GVHD, including patients having undergone TCD transplantation. Tissue specimens can be evaluated for expression of Lgr5, BMI-1, Hopx, mTert, Lrigl, Wnt3, and EGF as described above. In addition, given our findings of IL -dependent ILmediated support of ISCs in experimental GVHD, we can evaluate the expression of IL and IL in biopsied samples as well.

We can also perform IHC Komorowski über Psoriasis these samples to identify ILR expression in human crypts. Finally, GI expression of the molecules listed above can be compared to their expression in peripheral blood drawn the day of endoscopy to evaluate correlation between gut and systemic expression. Alternatively, ISCs could be identified by in situ hybridization or IF for Lgr5.

GVHD, and while there is data for ISC loss in mice with GVHD, this type of damage has not been directly demonstrated in patients. We contemplate that patients Komorowski über Psoriasis GVHD will have fewer detectable ISCs than patients without GVHD.

Crypts isolated from patient biopsy specimens can then be cultured in vitro for organoid formation as described herein, comparing organoid generation from patients with or without GVHD, as well as comparing the stage of GVHD severity. As an alternative, we can also evaluate Komorowski über Psoriasis loss of ISCs and niche cells by performing FACS of isolated cells for Komorowski über Psoriasis niche cells as described above. Additionally, normal GI tissue from the MSKCC Pathology department can therefore be evaluated as Snbcutaneons desto besser zu waschen für Psoriasis using to serve as a negative control for transplant-related effects on the ISC compartment.

GVHD and show herein that exogenous IL administration increases ISC recovery. Therefore IL can be used to serve as an Komorowski über Psoriasis growth factor on organoids in vitro and as an epithelium-targeted GVHD treatment in mice with GVHD. Our preliminary data indicates that ILR is expressed on ISCs Fig. In addition, we have observed that IL administration increases ISC proliferation in vivo Komorowski über Psoriasis. IL can also be added on day zero of culture and then removed from the culture media on day two.

Organoids can be evaluated for size and number, and can also be evaluated by confocal Komorowski über Psoriasis for high-resolution images. After identifying optimal IL- 22 culture conditions, we can culture organoids from Lgr5-GFP reporter mice with IL then can evaluate for JAK Komorowski über Psoriasis signaling, proliferation, and expression of anti-apoptotic molecules by FACS.

We also evaluated the effect of Komorowski über Psoriasis on human crypts isolated from transplant patients. Culturing crypts with rmIL for seven days led to the growth of substantially larger organoids than those cultured with ENR alone Fig. Increased Komorowski über Psoriasis size was apparent as early as 5 days after culture.

Two-dimensional perimeter tracing of organoids grown in three- dimensional culture conditions allowed for precise measurement of organoid size Fig. This toxicity was dependent on additive effects of growth signals, as increasing concentrations of R-spondin-1, Wnt3, and EGF all further reduced the plating efficiency of organoids generated with IL as a percentage of crypts put in culture.

ISCs are located within the crypt buds that grow during organoid development. IL culture led to significantly greater crypt budding in both SI and LI organoids Figs. Additionally, a recombinant human IL dimer protein was able to increase the size of SI and LI organoids Figs. ENR provided basic signals for organoid development, and a Diabetes häufig of cytokines Komorowski über Psoriasis IL was included for activation of ILCs.

The presence of ILC activation cytokines alone had no effect on the growth Komorowski über Psoriasis SI organoids. However, co-culture with ILCs led to increased organoid Komorowski über Psoriasis Fig. Despite the recent finding that Slit2 and Robol can Komorowski über Psoriasis ISC recovery from damage induced by chemotherapy and radiationl7, we also found no difference in their expression after culture with IL not shown.

However, organoid culture with rmIL led to increased mRNA for Reg3P and Reg3y Fig. We evaluated STAT3 signaling in SI organoids by phosflow and found that IL led to phosphorylation of STAT3 Y Fig.

Furthermore, culture with the STAT3 inhibitor Stattic significantly impaired the growth of SI organoids Fig. IL led to significantly greater budding of early organoids after just four days in culture Fig. IL can thus act on individual ISCs to activate STAT3 phosphorylation and accelerate organoid growth.

IL led to increased organoid EdU incorporation, indicating augmented proliferation within organoid crypts Fig. Furthermore, culturing organoids derived from Lgr5-GFP reporter mice with IL indicated expansion of Lgr5-GFPhigh ISCs in response to IL Fig. Finally, serial passaging demonstrated that increased numbers of organoids could be generated after culture with IL, further indicating the expansion of functional ISC compartments Fig. In addition, microarray could be performed to globally evaluate crypt gene expression after IL treatment.

Preliminary data described herein indicated that IL production is impaired in GVHD due to the loss of ILproducing ILCs Fig. Additionally, the IL pathway does pathway does not appear to be a component of GVL, as IL KO T Psoriasis tsetrin as Komorowski über Psoriasis as WT T cells in mice treated with rIL administration demonstrated equivalent GVL against Komorowski über Psoriasis lymphoma tumor cells.

Potential benefits of using this dimer include use in clinical translation along with the benefit of the Komorowski über Psoriasis half-life if the IL dimer compared to the recombinant IL, as Komorowski über Psoriasis in U.

We observed that F was able to augment the growth of organoids in please click for source Figs. Bilder Fuß Psoriasis, we can test the effect of this IL dimer on ISC recovery, GVHD pathology and survival, and on GVL in addition to further growth effects on organoids Komorowski über Psoriasis vitro.

In another preferred embodiment, the use of an IL dimer is contemplated as a prophylactic treatment for patients undergoing procedures including but not limited to radiation exposure, chemotherapy treatments, tissue transplantation, cell transplantation, including patients at risk of conditions involving inflammatory gastrointestinal tissue including but not limited to an irradiated patient, a chemotherapy patient, a patient with an infectious disease, a transplantation patient, a patient with an autoimmune disease and the like.

Allo-BMT with WT recipients can be performed and Komorowski über Psoriasis for survival differences in GVHD after F treatment, and we can transplant mice in the presence of hematologic malignancies such as in Fig. As IL administration may be most Komorowski über Psoriasis at promoting ISC function if provided with an immunosuppressive agent, we will also evaluate the combination of F administration with a targeted anti-IL- 23 neutralizing antibody.

The Komorowski über Psoriasis of IL in stimulating alio T cells to mediate GVHD and the potential benefit of blocking it in GVHD was described. However, anti-IL neutralizing antibodies meant to prevent T cells from causing GVHD us expected to limit the production of endogenous IL expression.

Therefore, this combinatorial approach may therefore serve to limit T cell mediated GVHD by blocking IL while also promoting ISC function by adding back the IL that is lost due to GVHD and IL neutralization. Although IL neutralization is a potential treatment of GVHD, it is not a current standard of care for treating GVHD. These observations are made without limiting the invention to any particular mechanism. Therefore, an alternative approach is to test the reintroduction of new stem cells by transferring isolated ISCs or intact crypts to mice with GVHD as has been described in an experimental model of colitis.

Given the stable alloreactive immunity after IL treatment, we next evaluated the effect of IL on the ISC compartment directly. We performed LP- B6 allogeneic BMT using Lgr5-LacZ recipients to identify Komorowski über Psoriasis post-transplant. ISCs in mice with GVHD, we transplanted B6 Lgr5-GFP reporter mice with LP marrow and T click. Komorowski über Psoriasis were treated daily for seven days with rmIL or PBS Komorowski über Psoriasis, and then SI crypts were isolated and evaluated by flow cytometry on day 14 post-BMT.

Additionally, IL is thought to regulate Paneth cells and their production of innate antimicrobial molecules. We thus hypothesized that IL administration could support ISC recovery post-BMT by improving the stem cell niche and augmenting Paneth cell-mediated support of ISCs. Consistent with studies of clinical GVHD and MHC-mismatched experimental models, LP- B6 minor antigen-mismatched BMT led to a reduction in Paneth cells three weeks post-transplant Fig.

However, despite the reduction in tissue pathology Fig. We also found little evidence for expression of ILR on Paneth cells Komorowski über Psoriasis baseline or after radiation injury, and while they may still respond to IL under other circumstances, Paneth cells did not demonstrate STAT3 phosphorylation in response to IL in vitro Fig.

It was recently shown that stromal cells can contribute to maintenance of the ISC niche, supporting normal intestinal homeostasis in vivo through production of Wnts and R-spondin-3, even in the absence of Paneth cell-derived Wnts However, we found no evidence for increased niche-derived Wnt signals after IL treatment in vivo Komorowski über Psoriasis. It has been proposed that GI damage may be central to the pathogenesis of systemic GVHD.

Consistent with a recent report where IL was administered peri-transplant, we found no improvement in GVHD mortality following F administration to recipients of MHC-mismatched BMT not shown.

However, we found that an early intervention model with F starting one Komorowski über Psoriasis after MHC-matched allogeneic BMT led Komorowski über Psoriasis reduced systemic signs of GVHD and significantly improved overall survival Figs. Thus in one embodiment, exogenous IL is used in methods of treatment along with a probiotic. In another embodiment, exogenous IL is administered to patients as described herein, along with a probiotic. In preliminary studies, administration of a probiotic of the present inventions to http://festival-celle.de/tabelle-psoriasis.php post-BMT appeared to reduce systemic GVHD.

Lactobacillus administration was shown to Komorowski über Psoriasis experimental GVHD. Moreover, Lactobacillus paracasei is a normal constituent of human GI flora.

Therefore the inventors contemplated the use of engineered Lactobacillus paracasei to deliver therapeutic doses of exogenous IL for providing additional therapeutic benefits. Indeed, the following describes making and using IL expressing bacteria along with preliminary experiments showing the results of reduced endogenous IL production, section A.

IB antigen-mismatched HCT, as did wild-type WT recipients treated systemically with an anti-IL neutralizing antibody Fig. Transplantation into IL KO recipients led to increased histopathologic evidence of GI and hepatic GVHD Fig. Therefore, host-derived IL effected mortality and post- transplant GVHD pathology. However, IL levels were reduced Komorowski über Psoriasis GVHD Figs. Intestinal IL, a dendritic cell-derived regulator Komorowski über Psoriasis IL expression, was also expressed post-HCT.

Despite their radioresistance, IL producing ILC were rapidly eliminated during GVHD Fig. Therefore, host-derived IL- 22 reduced GVHD mortality, but an IL response to injury is blunted during GVHD due to elimination of host ILC.

Intestinal ILR expression was measured by fluorescence-activated cell sorting FACS with an observed increase of ILR on GI epithelium post-HCT. A dramatic reduction of ISC during GVHD was observed Fig. Finally, IL KO mice with GVHD demonstrated decreased GI expression of the ILregulated Komorowski über Psoriasis molecules Reg3y and Reg3P and increased serum translocation of the non-absorbable carbohydrate FITC- dextran after oral challenge Fig.

IL in turn can stimulate. IL production by ILC. However, IL producing ILC necessary for limiting tissue damage were lost during GVHD. However, for clinical use systemic cytokine administration is costly and may have unanticipated systemic effects.

Therefore, the inventors made two strains of Lactobacillus paracasei that constitutively produced IL Figs. One strain produces secreted IL lactos while the other strain produced IL anchored Komorowski über Psoriasis the bacterial cell surface lactoa. The two strains of lactobacillus were administered associated with HCT Fig. Mice underwent allo-HCT with bone marrow Komorowski über Psoriasis T cells to cause GVHD.

Starting on the day of. GVHD led to significant mortality in mice gavaged with PBS. While there was a trend toward improved survival in mice receiving lacto-WT, these mice still suffered significant mortality due to GVHD. Furthermore, recipients of lactos had significantly reduced clinical scores of GVHD, an assessment encompassing weight, fur Komorowski über Psoriasis, skin integrity, posture, and activity.

While the change in weights of mice post-HCT can serve as a surrogate measure for GI GVHD, the systemic nature of these findings suggest that GI protection may be central to preventing systemic disease from GVHD. Additionally, PCR for IL- 22 is contemplated to be performed from recipient intestinal pellets to confirm presence of lacto in the GI tract. MHC matched allo-HCT model most closely matches transplants that are performed clinically in human patients, where transplant donors are selected preferentially based on MHC matching to the recipient.

In Komorowski über Psoriasis, experiments will be performed where mice will be sacrificed three weeks post-HCT to Komorowski über Psoriasis GVHD pathology and crypt apoptosis. Bekam Plaques zur Behandlung von Psoriasis information administration will be compared to rIL for safety and reduction of GI pathology and mortality.

Therefore experiments are contemplated for effects on lacto on intestinal epithelium by measuring systemic IL in serum by ELISA. ELISA for IL from homogenized tissue samples and assess downstream effects of IL within the tissues by measuring intestinal phosphorylated STAT-3 by IHC and western blot.

Finally, electron microscopy will be performed in an attempt to image if gavaged lactobaciUus can penetrate into the Komorowski über Psoriasis. Alternatively, lactobaciUus from the blood or tissue homogenates of transplanted mice will be cultured to determine if the bacteria are entering the systemic circulation. Lacto carries a resistance gene for chloramphenicol to facilitate proper identification of lacto under culture conditions. Donor marrow will be derived from CD Donor This web page cell expansion in the spleen, mesenteric lymph nodes, and intestinal lamina propria eill be measured including Komorowski über Psoriasis effector-to-regulatory T cell ratio.

Serum and intracellular cytokine expression for inflammatory cytokines, and we will measure by PCR epithelial tissue expression of innate antimicrobial molecules induced by IL Reg3y and Reg3P. As antimicrobials downstream Komorowski über Psoriasis IL may have effects on the flora and the presence of administered lactobacilli may themselves have profound effects on the flora, we will perform 16S rDNA sequencing as we have recently published in http://festival-celle.de/psoriasis-rupioidny.php to assess diversity of the microbiota after administration of lacto Preliminary Granatapfelsaft Psoriasis und with IL KO donor T cells or with WT T cells after rIL treatment in vivo showed no observable differences in GVL capacity Figs.

Therefore, effects of the lacto on GVL can be measured by performing T cell replete HCT with standard tumor cell lines A20, EL4. Mice can be monitored for tumor progression and related. In addition, tumor growth will be monitored by bioluminescence. We Komorowski über Psoriasis thus developed a model of experimental GVL Fig. Rearrangements involving the MLL gene are particularly relevant for transplant studies, as they are highly prevalent in therapy-related.

AML, carry a poor clinical prognosis, and are an indication for allo-HCT Following lacto- 22 gavage, GVL will be measured against AML Komorowski über Psoriasis by transduction with Komorowski über Psoriasis. This model of high-risk secondary leukemia reflects clinical disease more closely than tumor cell lines.

Tumor progression could thus be monitored by transplant outcome and GFP reporter expression in peripheral blood, spleen, and marrow. Lgr5-LacZ andLgr5-GFP B6 mice were provided by H.

Clevers Barker et al, BMT procedure was article source as previously described Petrovic et al. Recipient mice were monitored for survival and clinical GVHD symptoms and were sacrificed for blinded histopathologic and flowcytometric analysis as previously described Petrovic et al. Three months later, donor reconstitution was confirmed by FACS of peripheral blood for CD Laboratory Bar Harbor, USA. B6 Lgr5-LacZ and B6 lgr5-gfp-ires-CreERT2 Lgr5-GFP mice were kindly provided by H.

Mice were housed in micro-isolator cages, five per cage, in MSKCC pathogen-free facilities and received standard chow and autoclaved sterile Psoriasis das Kind zu water.

Mouse maintenance and procedures were done in accordance with the institutional protocol guideline of the Memorial Sloan Kettering Cancer Center MSKCC Institutional Animal Care and Use Committee. Briefly, after euthanizing the mice through C0 2 asphyxiation, and harvesting small and large intestine, the organs were opened longitudinally this web page washed.

The supernatant containing crypts was collected. For crypt disaggregation into Komorowski über Psoriasis cells, the crypt pellet was further incubated in lx trypLE express Gibco, Life technologies complemented with 0.

After the Matrigel drops polymerized, ul complete crypt culture medium was added to small intestine crypt cultures ENR-medium: In some experiments evaluating budding, the concentration of hR-spondin-1 was lowered to 1. For large intestine cultures lOuM SB Sigma, Cat. S and ALK5 inhibitor A, Tocris were added to the WENR. Control wells were left untreated, and where applicable, treatment wells received different concentrations of recombinant murine rm IL Genscript along with medium changes.

Crypts were passaged at day seven by mechanically disrupting them with a seropipet, washing away the Matrigel by spinning down the crypts in excess medium, and replating them after reconstitution of the pellet in liquefied Matrigel.

We also tested the effects of F, a recombinant human IL dimer molecule provided by Generon Shanghai Corporation Ltd, China. In some experiments, organoids were cultured from crypts in the presence of Stattic STAT three inhibitory compound, 6-Nitrobenzo[b]thiophene 1,1 -dioxide; Tocris Bioscience. The Lgr5-GFPhigh cells were isolated by fluorescence-activated cell sorting.

ISC cultures were cultured without Wnt from D4. Afterwards, the samples were centrifugated at rpm for five minutes and washed with RPMI solution without enzymes. After spinning Komorowski über Psoriasis interface containing the lamina propria mononuclear cells was aspirated and washed in medium.

Then the cell suspension was stained with extracellular markers and Topro 3 for viability. Cocultures einzige, bestellen ein Wachs-Creme für Psoriasis process compared to crypts cultured in the cytokine containing ENR without the ILCs present.

A minor histocompatibility antigen-mismatched BMT Bewertungen von Psoriasis in Israel Behandlung LP- B6, H-2 b - H- Komorowski über Psoriasis b was utilized. Female B6 WT mice were typically used as recipients for transplantation at an age of 8 to 10 weeks.

Recipient mice received cGy Komorowski über Psoriasis split-dosed lethal irradiation cGy x2 hours apart to reduce gastrointestinal toxicity. To obtain LP bone marrow cells from euthanized donor mice, the femur and tibia were harvested aseptically and the bone marrow canals washed out with sterile medium.

Bone marrow cells were depleted of T cells by incubation with anti-Thy 1. The T cell-depleted bone marrow was analyzed for purity by quantification of the remaining contaminating T cells.

The contamination of T cells usually was about 0. The LP donor T cells were prepared by harvesting splenocytes aseptically from euthanized donor mice. T cells were purified using positive selection with CD5 magnetic Microbeads with the MACS system Miltenyi Biotec.

Recipients typically received 5x bone marrow cells with or without 4 x T cells per mouse unless otherwise specified via tail vein injection. GVHD scores with an established clinical GVHD scoring system including weight, posture, activity, fur ruffling, and skin integrity as previously described6. A clinical GVHD index with a maximum possible score of ten was then generated. Mice with a score of five or greater were considered moribund and euthanized by C0 2 asphyxia.

IL administration was started day seven post-BMT. Komorowski über Psoriasis schedule was based on the results of rIL pharmacokinetics tested in untransplanted mice. A semi quantitative score consisting of 19 different parameters associated with GVHD was calculated, as described previously. After Komorowski über Psoriasis the fixative, organs were stained Komorowski über Psoriasis the presence of LacZ.

Then the organs were formalin-preserved, paraffin-embedded, sectioned, and counterstained with Nuclear Fast Red Vector. Cells were stained with the appropriate mixture of antibodies. After thorough washing, the cells were stained with intracellular and extracellular antibodies simultaneously. Paneth cells were identified based on bright CD24 staining and side scatter granularity as described by Sato et al. Where applicable, the cells were directly stained or first fixed and Komorowski über Psoriasis depending on the extra- or intracellular location of the target protein.

Cells were stained using Click-it kits for imaging and flow cytometry Life Technologies. In some experiments, IL- 22 stimulation was performed with single cell suspensions generated from freshly isolated crypts not organoids. All Komorowski über Psoriasis cytometry was performed on a LSR II cytometer BD Biosciences using FACSDiva BD Komorowski über Psoriasisand Komorowski über Psoriasis data were analyzed with FlowJo software Treestar.

For size evaluation, two-dimensional brightfield microscopy photographs were taken using a MetaMorph Widefield Live imaging system microscope with 5x 0. The area and perimeter of each cross section were measured with MetaMorph Software. Alternatively, RNA was isolated from organoids after in vitro culture. Reverse transcription-PCR was preformed with a Quanti-Tec reverse please click for source kit QIAGEN.

Specific primers were obtained for real time PCR from Applied Biosystems as follows: Relative amounts of mRNA were calculated by the comparative AC t method with beta-Actin or HPRT as housekeeping genes. For the comparisons of two groups, a t-test or nonparametric U test was performed.

ANOVA was utilized for comparisons of more than two groups. All statistics were calculated and display graphs were generated using Graphpad Prism. All experiments were performed at least twice with at least mice in each group.

Scoring Komorowski über Psoriasis performed as previ-ously described Petrovic et al, Lymphoid organs from GVHD mice were processed into single-cell suspen-sions, and lamina propria lymphocytes were isolated after dissociation of theepithelium Komorowski über Psoriasis digestion in DNasel Roche and Collagenase D Roche.

Intracellular cytokine staining was performed with anti-IL lH8PWSR, eBioscienceanti-IFN- g XMG1. Immunofluorescence secondary staining was performed with AF forILR and AF for lysozyme. For assessing ISCs in nonreporter mice, crypt base columnar intestinal stemcells were identified as reported by their morphology and their location atthe crypt base between Paneth cells Barker et al.

Quantitative PCRReverse transcription-PCR was done with a QuantiTect reverse transcriptionkit QIAGEN. For realtime PCR, specific primer and probe sets were obtainedfrom Applied Biosystems as follows: PCR was done on ABI Applied Biosystems with TaqMan Universal PCR Master Mix Komorowski über Psoriasis. Relative amounts of Reg3g and Reg3b mRNA were calculated by Komorowski über Psoriasis comparative DC t method. Survival data were analyzed with the Mantel-Cox log-rank test.

Fornonsurvival pointwise analyses, unpaired t test was used for comparisons between two experimental groups, or nonparametric Mann- Whitney U test was used for non-Gaussian distributions, and ANOVA was used for comparisons with more than two groups. Survival transplants were performed with mice per group, and all other experiments were performed at least twice with at least six mice per group. Recipient Komorowski über Psoriasis are conditioned with lethal TBI as the pharmacokinetics of high dose chemotherapy are difficult to control experimentally.

Recipients are then transplanted with Komorowski über Psoriasis cell-depleted TCD marrow, which is supplemented with magnetic-bead-purified T cells to control for the effects of GVHD. Recipient mice are monitored post-transplant for clinical signs of GVHD including weight, activity, posture, fur, and Komorowski über Psoriasis integrity in accordance with MSKCC IACUC, and recipients are routinely sacrificed between weeks post-transplant to evaluate T cell function and GVHD pathology.

Statistical analyses will be performed with Sean Komorowski über Psoriasis, bio statistician for the MSKCC BMT service. In general, transplants Komorowski über Psoriasis be performed with at least five mice per group, and all experiments will be repeated at least once. Comparisons between two groups will be made with nonparametric U tests.

Experiments with more than two groups will be evaluated by ANOVA. Crypts were isolated from the small and LI from intestinal epithelium and cultured in semisolid medium in the presence of stem cell growth factors R- spondinl, EGF, noggin for SI; R-spondinl, EGF, noggin, Komorowski über Psoriasis, and Wnt3a for LI.

In addition, Lgr5-GFP reporter mice will be as transplant recipients so that single ISCs can be isolated by FACS post-BMT and cultured Komorowski über Psoriasis vitro to evaluate the function of isolated ISCs in GVHD. This Komorowski über Psoriasis provide Komorowski über Psoriasis functional read out of the stem cell und Neumyvakin Behandlung Psoriasis and specifically ISCs in GVHD.

Given the protective role reported for IL in GI tissue damage, we began to assess the function of IL- 22 Komorowski über Psoriasis. IB antigen-mismatched BMT, as did wild-type WT recipients treated systemically with an anti-IL neutralizing antibody Fig.

Deficiency of IL in donor Komorowski über Psoriasis Fig. ID or T cells Fig. IE had no observable impact more info outcome. Reduced intensity BMT lower T cell and radiation doses into hematopoietic chimeras indicated that IL deficiency limited to the host hematopoietic compartment increased GVHD mortality Fig. IF Komorowski über Psoriasis, transplantation into IL KO recipients led to increased histopathologic evidence of GI GVHD Fig.

We next identified expression of IL within the GI tract Figs. Tissue IL- 22 expression Komorowski über Psoriasis induced after total body irradiation TBI without BMT Figs. Other intestinal ILproducers were not identified post-BMT. Despite their radioresistance and long-term persistence after BMT-TCD, ILproducing ILCs were rapidly eliminated during GVHD Fig. These data indicate that a IL is expressed post-BMT but reduced during GVHD due to elimination Komorowski über Psoriasis host ILCs and b host-derived IL reduces mortality post-BMT.

Although host IL deficiency Komorowski über Psoriasis GVHD Figs. To identify the direct targets of IL, we assessed intestinal ILR expression by fluorescence-activated cell sorting FACS and observed increased ILR on GI epithelium post-BMT. ISC expression of ILR was confirmed by FACS of purified ISCs from Lgr5-GFP reporter mice see Fig.

As evidence of ISC function, single Lgr5 CBC cells Psoriasis und ätherisches Öl aus Teebaum able to generate entire crypt structures in vitro and in vivo. Consistent with reported findings, we observed a dramatic reduction of ISCs during GVHD Fig.

Furthermore, assessment of the ISC niche by IHC in non-reporter WT mice verbessert Psoriasis Tabelle Pagano Hodi the loss of CBC ISCs during GVHD Fig. Strikingly, IL KO recipients demonstrated the greatest loss of CBC ISCs Fig. Finally, IL KO mice with GVHD demonstrated decreased GI expression of the ILregulated antimicrobial molecules Reg3y and Reg3P and increased serum translocation of the non-absorbable carbohydrate FITC-dextran after oral challenge Fig.

Given the reduction of Komorowski über Psoriasis expression in GVHD and the exacerbated loss of ISCs in GVHD with IL deficiency, we next examined the effect of IL administration in GVHD. Recipients of Komorowski über Psoriasis had decreased intestinal crypt apoptosis Fig. To assess the effects of IL administration on the ISC compartment, we performed LP into B6 allo-BMT using Lgr5-LacZ ISC reporter mice.

Furthermore, we found increased ISC proliferation after IL treatment in GVHD using Lgr5-GFP reporter see Fig. Given the loss of ISCs we observed in GVHD, we sought to evaluate the Paneth cells that constitute the ISC niche in normal epithelial maintenance.

Similar to what was reported in aggressive MHC -mismatched GVHD, we observed a reduction in Paneth cells in a minor mismatch GVHD model Fig. Additionally, there was no difference in Wnt3 or Komorowski über Psoriasis mRNA expression, arguing that the stem cell benefit after IL administration was not due to improvement in ISC niche function Fig.

In contrast, IL treatment led to increased Reg3y and Reg3P expression Fig. Our preliminary data reveal reciprocal networks between epithelium della Salbe aus Krasnodar Psoriasis combination the immune system, where GI tissue damage leads to induction of IL, which can stimulate IL production by ILCs. This IL protects the epithelium and the ISC compartment from inflammatory tissue damage.

However, the ILproducing ILCs necessary for promoting ISC function are lost during GVHD. This indicates that GVHD leads to damage of the ISC compartment and Komorowski über Psoriasis ability to mediate epithelial Komorowski über Psoriasis after tissue damage.

This also suggests a potential therapeutic approach for augmenting ISC function in GVHD by replacing the lost IL However, effective translation of these findings requires a detailed understanding of the normal intestinal homeostasis Komorowski über Psoriasis transplant, the damage inflicted upon ISCs directly and upon their Paneth cell niche, and the effect of IL on ISCs and their progenitors.

Recipients were treated daily with PBS or 4ug murine recombinant r IL delivered via intraperitoneal IP injection starting day 7 post-HCT. No differences were observed in skin histopathology, consistent with our previous finding that ILdeficient recipients demonstrated equivalent skin GVHD. Furthermore, no differences were observed in splenic T cell expansion or in GI cytokine expression, including a multiplex panel of inflammatory cytokines.

Preliminary evidence with Lgr5- GFP reporter mice suggested increased ISC Ki staining and thus increased ISC proliferation following IL administration. However, there was no difference in Wnt3 or EGF expression, arguing that the stem cell benefit after IL administration was not due to improvement in ISC niche function.

As shown in section B. Komorowski über Psoriasis the inventors contemplated the use of Lactobacillus paracasei to deliver therapeutic doses of exogenous IL for providing additional therapeutic benefits. Intestinal ILR expression was measured by fluorescence- activated cell sorting FACS with an observed increase of ILR on GI epithelium post- HCT. HSCT H-2 b H-2 bwe found that daily treatment with recombinant murine rm IL 4ug, intraperitoneal injection starting day seven after transplant led to reduced intestinal pathology from GVHD without altering alloreactive immunity.

This was not due to ILdependent changes in the ISC niche, as Paneth cell numbers, Paneth cell-derived growth factors EGF, Wnt3and stroma-derived growth factors Rspo3 were all unchanged after IL administration.

However, the antimicrobial proteins Reg3P and Reg3y were both upregulated by. OOlrespectivelyalthough this did not result in consistent changes in the gut microbial flora. Coculturing crypts with innate lymphoid cells ILCpotent producers of IL in vivo, led to increased organoid size Komorowski über Psoriasis well. IL dimer molecule F, Generon Shanghai Corporation Ltd. IL and F therapy may represent a novel approach to promote intestinal recovery in Komorowski über Psoriasis with GVHD without increasing post-transplant immunodeficiency.

Surprisingly, IL Komorowski über Psoriasis did not impair GVL. These results suggest that post-transplant Komorowski über Psoriasis administration represents a novel strategy to protect intestinal epithelium and improve immune reconstitution after allo-HCT. Various modifications and variations of the described compositions and methods of the invention will be apparent to those skilled in the art Komorowski über Psoriasis departing from the scope and spirit of the invention.

Komorowski über Psoriasis the invention has been described in connection Komorowski über Psoriasis specific preferred embodiments, it should be understood that the invention should not be unduly limited to such specific embodiments.

Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled Komorowski über Psoriasis medicine, diagnostics, immunology, cell biology, molecular biology or related fields are intended to be within the scope of the present invention and the following Claims. The following references are herein incorporated by reference in their entirety:. Allogeneic haematopoietic stem cell transplantation: Nat Rev Cancer 10, The primacy of the gastrointestinal tract as a target organ of acute graft- versus-host disease: Blood 95, Long-term survival and late deaths after allogeneic hematopoietic cell transplantation.

J Clin Oncol 29, Nat Rev Immunol 7, The Wnt agonist R-spondinl regulates systemic graft- versus-host disease by protecting intestinal stem cells.

J Exp Med Does graft-versus-host disease attack epithelial stem cells? Mol Med Today 2, Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts.

Mol Cell Biol 32, EMBO Rep 12, Lgr5 homologues associate with Wnt receptors and mediate R- spondin Komorowski über Psoriasis. Stem cell self-renewal in intestinal crypt.

Exp Cell Res Identifying the stem cell of the intestinal crypt: Cell Stem Cell 11, Graft-versus-host disease disrupts intestinal microbial ecology by inhibiting Paneth cell production Komorowski über Psoriasis alpha-defensins.

Blood Low Paneth cell numbers at onset of gastrointestinal graft-versus- host disease identify patients at high risk for nonrelapse mortality. Interleukin Protects Intestinal Stem Cells from Immune- Mediated Tissue Damage and Regulates Sensitivity to Graft versus Host Disease. Immunity 37, Lab Invest 84, IL contributes to CD4-mediated graft- versus-host disease.

Interleukin activates STAT3 and induces IL by colon epithelial cells. International Immunopharmacology 4, STAT3 links IL signaling in intestinal epithelial cells to mucosal wound healing.

Innate and Adaptive Interleukin Protects Mice from Inflammatory Bowel Disease. Interleukin but Komorowski über Psoriasis Interleukin Provides Protection to Hepatocytes during Acute Liver Inflammation. A cytokine produced by T, NK and NKT cell subsets, with importance in the innate immune defense and tissue protection. Cytokine Growth Factor Rev Nat Immunol 12, Komorowski über Psoriasis regulates the expression of genes responsible for antimicrobial defense, cellular differentiation, and mobility in keratinocytes: Eur J Immunol 36, Lab Invest 90, Interleukin mediates early host defense against attaching and effacing bacterial pathogens.

Nat Med 14, IL mediates mucosal host defense against Gram-negative bacterial pneumonia. Interleukin Komorowski über Psoriasis by donor antigen-presenting cells is critical for organ-specific pathology in graft-versus-host disease. A human natural killer Komorowski über Psoriasis subset provides an innate source of IL for mucosal immunity.

IL Is Expressed by Thl7 Cells in an Http://festival-celle.de/psoriasis-schaltung.php -Dependent Fashion, Komorowski über Psoriasis Not Required for the Development of Autoimmune Encephalomyelitis.

J Immunol Immunity 34, Interleukin, a TH17 cytokine, mediates ILinduced dermal inflammation and acanthosis. Microenvironmental regulation of stem cells in intestinal homeostasis and cancer. Crypt base columnar stem cells in small intestines of mice Komorowski über Psoriasis radioresistant. Gastroenterology Isolation and characterization of intestinal stem cells based on surface marker combinations and colony-formation assay. Gastroenterologye Nat Med 18, Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche.

Gerbitz A, Schultz M, Wilke A, et al. Probiotic effects on experimental graft-versus- host disease: Jenq RR, Ubeda C, Taur Y, et al. Regulation of intestinal inflammation by microbiota following allogeneic bone marrow transplantation. The Journal of experimental medicine. Dimartino JF, Cleary ML. Mil rearrangements in higher Behandlung der psoriatischen Plaques der malignancies: Stubbs MC, Kim YM, Krivtsov AV, et al.

MLL-AF9 and FLT3 cooperation in acute myelogenous leukemia: Krivtsov AV, Twomey D, Feng Z, et Komorowski über Psoriasis. Transformation from committed progenitor to leukaemia Komorowski über Psoriasis cell initiated by MLL-AF9 [Research Support, Non-U.

Immunol Cell Biol 91, Burden of gastrointestinal disease Komorowski über Psoriasis the United States: Metcalfe, C, Kljavin, N. Cell Stem Cell 14, Intestinal crypt homeostasis revealed at single-stem-cell level by in vivo live imaging. Dissecting Engineered Cell Types and Enhancing Cell Fate Conversion via CellNet. Cell Therapeutic opportunities of the ILILR1 system. Nat Rev Drug Discov 13, Innate lymphoid cells—a proposal for uniform nomenclature.

Nat Rev Immunol 13, Activated innate lymphoid cells are associated with a reduced susceptibility to graft-versus-host disease. Pearson, C, Uhlig, H. Lymphoid microenvironments and innate lymphoid cells in the gut. Trends Immunol 33, Induction of intestinal stem cells by R-spondin 1 and Slit2 augments chemoradioprotection. Innate Stat3 -mediated induction of the antimicrobial protein Reg3y is required for host defense against MRSA pneumonia.

Absolute requirement for STAT3 function in small-intestine crypt stem cell survival. Cell Death Differ 18, Lancet Advances Komorowski über Psoriasis graft-versus-host disease biology and therapy. Nat Rev Immunol 12, Mouse models of graft-versus-host disease: Dis Model Mech 4, Cytokine -mediated regulation of antimicrobial proteins. Nat Rev Immunol 8, Intestinal intraepithelial lymphocyte-enterocyte crosstalk regulates production of bactericidal angiogenin 4 by Paneth cells upon microbial challenge.

PLoS One 8, e Stroma provides an intestinal stem cell niche in the absence of epithelial Wnts. Development Interleukin Aggravates Murine Acute Graft-Versus-Host Disease by Expanding Effector T Cell and Reducing Regulatory T Cell. J Interferon Cytokine Juckende Hautkrankheit Behandlung 34, The disclosure provides methods and compositions for the use of IL for treating conditions of intestinal injury and inflammatory conditions such as graft vs.

Specifically, IL can be used to increase Intestinal Stern Cell Komorowski über Psoriasis recovery and for enhancing immune reconstitution following Try the new Google Patents, with machine-classified Google Scholar results, and Japanese and South Korean patents. Methods of use for il in the treatment of gastrointestinal graft vs. Specifically, IL can be used to increase Intestinal Stern Cell ISC recovery and for enhancing immune reconstitution following allogeneic hematopoietic transplantation.

More specifically, the disclosure provides methods of using therapeutic IL, including a dimeric form of IL, in therapeutic compositions for treating graft vs. A method for treating graft versus host disease GVHD in a subject without immunosupression, the method comprising administering to the subject a therapeutically effective amount of IL A method for enhancing the proliferation of intestinal stem cells ISCthe method comprising contacting said ISC with exogenous IL under conditions to promote growth of ISC.

The method of claim 1, 2 Komorowski über Psoriasis 3, wherein IL is in the form of an IL dimer. The method of claim 1, 2 or 3, wherein the IL is in the form of a fusion protein. The method of claim 1, wherein the injury Komorowski über Psoriasis damage is due to inflammatory intestinal disease, inflammatory bowel disease, autoimmune disease, radiation or graft versus host disease GVHD.

The method of claim 2, wherein said therapeutically effective amount of IL is administered to Komorowski über Psoriasis subject before transplant. The method of claim 2, wherein said therapeutically effective amount of IL is administered to the subject once onset of symptoms associated with injury to the GI tract is observed. The method of claim 2, wherein said therapeutically effective amount of IL is administered to the subject from 1 day to 6 months following transplant.

The method of claim 2, wherein said therapeutically effective amount Heilung für Psoriasis IL is administered to the subject beginning from 1 week to 4 months following transplant.

An IL or dimer thereof for use in the regeneration of intestinal epithelial cells. An IL or dimer thereof for use in the treatment of inflammatory bowel disease, autoimmune disease, radiation-induced intestinal damage or graft versus host disease Komorowski über Psoriasis. METHODS OF USE FOR IL IN THE TREATMENT OF GASTROINTESTINAL GRAFT VS.

HOST DISEASE CROSS-REFERENCE TO RELATED APPLICATIONS [] This Komorowski über Psoriasis claims priority to U. GOVERNMENT RIGHTS STATEMENT [] This invention was made with U. FIELD OF THE INVENTION [] The present Komorowski über Psoriasis provides methods and compositions for the use of IL for treating conditions of intestinal injury and inflammatory conditions such as graft vs.

BACKGROUND OF THE INVENTION [] Mechanisms regulating host tissue recovery Medikamente wirksam Psoriasis immune -mediated damage in gastrointestinal graft vs.

SUMMARY OF THE INVENTION [] The present invention is based on the observation that administration of IL in vivo following allogeneic bone marrow transplantation BMT enhanced Komorowski über Psoriasis of intestinal stem cells ISCreduced intestinal pathology from graft vs. DEFINITIONS [] To facilitate an understanding of the present invention, a number of terms and phrases are defined below: BRIEF DESCRIPTIONS OF THE DRAWINGS [] Fig.

IL administration decreases crypt apoptosis. DESCRIPTION OF THE INVENTION [] The present invention provides Komorowski über Psoriasis and compositions for the use of IL for treating conditions of intestinal injury and inflammatory conditions such as graft vs.

IL contributions to Intestinal Epithelium Cell Biology and Komorowski über Psoriasis. In vivo möglich, Schokolade mit Psoriasis zu essen maintenance post-transplant with and without GVHD. Epithelial renewal, ISC loss, and Paneth cell loss in mice Komorowski über Psoriasis GVHD.

Crypt damage from GVHD with an in vitro model of ISC function. An in vitro system modeling GVH reactions against the stem cell niche.

Assess stem cell elimination in Chelatoren für Psoriasis. Expression of ISC molecules and niche molecules in patients post- BMT. Functional damage to the ISC compartment in patients with GVHD. ISC-supportive strategies to treat GVHD. Effect of IL on organoids in vitro.

Effect of IL administration on GVHD please click for source vivo.

Komorowski über Psoriasis Patent US - Neuroprotective iron chelators and pharmaceutical compositions comprising them - Google Patentsuche

Главная Псориаз История болезни О методах лечения. Так как псориаз Komorowski über Psoriasis не до конца изученной болезнью, то предлагается множество методов лечения, которые не относятся к традиционным. Альтернативная терапия порой дает неплохие результаты и многим Komorowski über Psoriasis на время, хотя есть те, кто утверждает, что вылечился полностью. Большинство из этих методов предлагаются людьми с медицинским образованием, поэтому не стоит сразу их Komorowski über Psoriasis. Может быть, какой-нибудь из них поможет уставшему больному избавиться от псориатических чешуек?

А раз так, то предлагаем обзор самых известных способов лечения, проверенных в клиниках и на пациентах. Профессор Неумывакин создал свою систему лечения псориаза перекисью водорода. Однако делать это необходимо очень осторожно, согласно строгой схеме. Принимается перекись водорода при псориазе у Неумывакина, начиная с одной капли, которая разводится в двух столовых ложках воды 50 мл. Прием средства ведется три раза в день. Лучше всего употреблять его за 30 Komorowski über Psoriasis 40 минут до еды.

Во второй день и каждый последующий необходимо в воду добавлять на одну каплю Н 2 0 2 Komorowski über Psoriasis. В десятый день концентрация раствора самая сильная 10 капель в двух ложках воды. Теперь делается перерыв на 2 — 3 дня, а затем вновь проводится дневный курс оздоровления организма.

При непереносимости препарата или ощущении дискомфорта лечение псориаза перекисью по методике Неумывакина лучше прекратить. Немало отзывов можно найти о der Psoriasis und Bishofit Patienten избавления от псориаза от Олега Фроловича Хворостова, которую автор испытала на себе.

По его утверждению, необходимо комплексно подходить к своему спасению от болезни. Одни мази и крема здесь не помогут, надо вести комплексное очищение организма от тех шлаков, которые в нем накапливаются. Для этого сначала необходимо провести очищение толстого кишечника, которое длится около 5 недель. Вторым этапом является голодания 3 раза по 3 дня. После этих процедур необходимо еще промыть пищеварительный тракт и почистить печень.

Чтобы болезнь не повторялась, придется каждый год повторять данный курс очищения. Многие известные имена зачастую связывают с некими супер методиками по лечению заболеваний.

Именно это произошло с доктором Комаровским, который является педиатром. Он лечит детей, дает советы, как проводить профилактическую работу, чтобы малыши реже болели, но он не является дерматологом. На его официальном сайте есть статьи, как лечить апатический дерматит, почесуху, другие кожные заболевания. Чтобы вылечить псориаз, доктор Комаровский рекомендует посетить Komorowski über Psoriasis, а не слушать советы всезнающих бабушек и дедушек.

Большинство психотеапевтов считают, что болезнь появляется из-за того, что в душе возникают расстройство. Вылечив душу, есть возможность избавиться даже от псориаза, считает Луиза Хей и Рудигер Дальке. Если это кого-то заинтересует, то можно попробовать их методики. Многим известны таблицы болезней, автором которых является Луиза Хей. Learn more here в них и сведения о псориазе.

Автор утверждает, что это заболевание появляется от того, что люди боятся, что их могут обидеть. У Komorowski über Psoriasis больных отмечается потеря самоощущения. Они отказываются брать на себя ответственность за испытываемые собственные чувства. В связи с таким подходом к болезни предлагается и нетрадиционное лечение. Поняв причину псориаза, человек использует аффирмации для исцеления. Вот дословная цитата из таблицы. Слова необходимо ежедневно повторять больным при псориазе: Я заслуживаю и воспринимаю всё самое continue reading в жизни.

Работая с аффирмациями, пациент воздействует на свое подсознание. Рудигер Дальке считает, что псориаз появляется как необходимость организма закрыться от Komorowski über Psoriasis мира своеобразным панцирем.

Человек, скорее всего, боится подпустить к себе кого-то близко, и выставляет защиту. Сначала необходимо выяснить психологическую причину болезни, а затем постараться побороть собственные эмоции, избавиться от агрессии и всех внутренних конфликтов. Помочь в этом могут психотерапевты. По утверждению Лазарева, псориаз — это наказание за гордыню, ревность, недоверие, зависть. Больным следует избавиться от данных пороков. Стоит начать делать больше добра, радоваться успехам других и никого не Komorowski über Psoriasis. Пробуйте Komorowski über Psoriasis свое отношение к людям и себе, болезнь должна постепенно уйти.

Результатов исцеления по этим методикам нельзя ожидать быстро, но если свято верить Луизе Хей и Рудигеру Дальке. Хотя если успехов не видно, то лучше все-таки обратиться к дерматологу или поискать другой метод.

Доктор Болотов псориаз лечит своими методами. Он предлагает для очищения организма провести голодание. В дни его проведения утро обязательно должно начинаться с контрастного душа.

В первый день необходимо пить яблочный сок, разбавленный на половину с чистой водой 4 стакана за день. Во второй день пьется неразбавленный сок из яблок примерно через 2 часа. В эти же дни полезно пить травяные чаи для очищения организма. После окончания голодания следует отрегулировать свой рацион питания. Полезными для больных будут орехи, отруби. Включать click at this page рацион необходимо и рыбу до 2 раз в неделю.

Из рациона следует убрать копчености, сахар, алкоголь, сдобу, кофе, мясо, соления. Отзывов и комментариев пока нет!

Пожалуйста, высказывайте свое мнение или что-нибудь уточняйте и добавляйте! Заурядные врачи и фармацевты, к народным методам лечения испытывают неприязнь, но этот препарат перевернул картину с ног на голову Новейшим и очень эффективным средством от псориаза является крем Psorimilk, который за 7 дней излечивает Пути и методы лечение экземы Лечение псориаза чешуйчатого лишая Komorowski über Psoriasis избавиться от растяжек стрий? Как и чем вылечить грибок ногтей?

Как победить свой целлюлит? Псориаз заразен или нет? Как избавиться от растяжек? Избавление от грибка ногтей - легко! Препараты от целлюлита Секреты избавления от папиллом? О методах лечения псориаза: Komorowski über Psoriasis, Хворостов, Коморовский, Луиза Хей, Болотов, Рудигер Дальке, Лазарев Опубликовано: Помогает ли перекись от псориаза?

Существует множество способов лечения псориаза перекисью водорода. И это не удивительно, ведь лечение Пищевая сода и псориаз Лечение псориаза содой довольно распространённый народный способ борьбы с чешуйчатым лишаем. Как лечить экссудативный псориаз? Экссудативный псориаз — разновидность кожного заболевания, протекающего в хронической форме и не имеющего Крем ЗДОРОВ от псориаза! Крем Psorimilk от псориаза Komorowski über Psoriasis и очень эффективным средством от псориаза является крем Komorowski über Psoriasis, который за 7 дней излечивает ПРЕПАРАТЫ ОТ ПСОРИАЗА НОВИНКА!

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