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New Cytokines CE ELISA kits Launching File Part5: Hematopoiesis-Differentiation Products: Format: Size: cat# Product Description KAPMS120 human IL-5 KAPMS113 human IL-13 KAPMS105 human TGF-B ELISA 96 Tests Table of contents: 1. Overview 2. Hematopoiesis products 3. Technical Performances 4. Package Inserts Cytokines CE- Part5- Hematopoiesis-Differentiation June 2008 1/5 1. Overview Hematopoiesis is the process by which all the different cell lineages that form the blood and immune system are generated from a common pluripotent stem cell. During the life of an individual, two separate hematopoietic systems exist, both arising during embryonic development but only one persisting in the adult. 2. Hematopoiesis/Differentiation products - Clinical background 1. IL-5 (Interleukine-5) ELISA KIT Human IL-5 has been described as T-cell replacing factor and as B-cell growth factor II based on its induction of proliferation and Ig secretion by activated B cells. Several other activities have furthermore been attributed to this cytokine, including induction of growth and differentiation of eosinophils from bone marrow stem cells , induction of mIL-2R expression on activated B cells and enhancements of IgA secretion by LPS-stimulated Bcells . IL-5 has been shown to be produced by the TH2 subset of human TH clones . The spectrum of biological activities of IL-5, on B cells as well as other cell types suggests that IL-5 plays an important functional role in the activities of TH2 cells. IL-5 is a heavily glycosylated glycoprotein with a native m.w. of 45.000 to 60.000, with multiple subunits of between m.w. 24.000 and 28.000 under reducing conditions. 2. IL-13 (Interleukine-13)ELISA KIT Introduction Interleukin-13 (IL-13) was first described as a protein product, designated P600, encoded by an RNA produced by activated mouse Th2 cells . More recently, the cDNA cloning of the human homologue of P600, human IL13, has been reported . Human IL-13 is a nonglycosylated protein of 132 amino acids with a molecular weight of 12000Da. The IL-13 gene is located on chromosome 5q23-31, in the same region as the genes encoding IL-3, IL-4, IL-5, and GM-CSF [9]. It is produced by activated CD4 + and CD8+ T cells, but the expression seems to be more abundant in CD4+ T cells [5]. Although it is a Th2 cytokine in the mouse, it appears to be produced by Th0, Th1, and Th2-like human T cell clones. IL-13 is a pleiotropic cytokine whose spectrum of action encompasses B cells, mononuclear phagocytes, and large granular lymphocytes . IL-13 induces CD23 expression on B cells, promotes B cell proliferation in combination with anti-Ig or CD40 antibodies, and stimulates secretion IgE, and IgG4 [2,5,8]. IL-13 has also been shown to prolong survival of human monocytes and increases surface expression of MHC class II, CD23, and members of the integrin superfamily, like CD11b, CD11c, CD18, CD29 and CD49e . IL-13 inhibits the production of a series of cytokines like IL-1, IL-6, TNF- , and IL-8 by activated human monocytes . IL-13 induces IFN- production by NK cells. Clinical applications - The capacity of IL-13 to induce IgE synthesis indicates that IL-13 may play an important regulatory role in IgE-mediated atopic diseases. The measurement of IL-13 in body fluids may thus provide further information about the pathophysiology of atopic diseases. Furthermore, IL-13 inhibits HIV-1 replication in primary culture-derived macrophages and represents a candidate cytokine for the suppression of HIV infection within monocytes and macrophages in vivo. Cytokines CE- Part5- Hematopoiesis-Differentiation June 2008 2/5 3. TGF-1 (Transforming Growth Factor-beta 1)ELISA KIT Introduction Transforming growth factor- (TGF-) belongs to a family of dimeric 25 kDa polypeptides that are ubiquitously distributed in tissues and synthesized by many different cells . Three isoforms of transforming Growth Factor- (TGF-beta1, beta-2 and beta-3) exist in mammals. They play critical roles in growth regulation and development. Each isoform is encoded by a unique gene on different chromosomes. All three of these growth factors are secreted by most cell types, generally in a latent form, requiring activation before they can exert biological activity. The TGF-betas possess three major activities: they inhibit proliferation of most cells, but can stimulate the growth of some mesechymal cells; they exert immunosuppressive effects and they enhance the formation of extracellular matrix. Two types of membrane receptors possessing kinase activity are involved in signal transduction. The TGF-betas are involved in wound repair processes and in starting inflammatory reaction and then in the resolution through chemotactic attraction of inflammatory cells and fibroblast. TGF-1 is the first recognized transforming growth factor, its subunits of each 12.5 kDa are bound via disulphide bridges. TGF-1 is inhibitive to T- and B cell proliferation as well as to maturation and activation of macrophages. It furthermore inhibits activity of natural killer cells and lymphokine activated killer cells and blocks production of cytokines. Measurement of TGF-1 in blood has been advocated for diagnosis of various diseases. TGF-1 has been shown to be an organizer of responses to neurodegeneration . Clinical applications In this context, it turned out to be interesting in monitoring Alzheimer’s disease , Down’s syndrome, AIDS and Parkinson’s disease. Serum and cerebrospinal fluid levels of Multiple Sclerosis patients were shown to be of great value to monitor remission and acute phases . TGF-1 is thought to play an important role in bone metabolism, it is considered a putative regulator of osteoclastic-osteoblastic interaction, and thus it can be regarded as a marker for osteoporosis . TGF-1 is involved in the pathogenesis of glomerular diseases such as diabetic nephropathy and glomerulosclerosis. TGF-1 has been described to be functionally connected to major immune system abnormalities as in autoimmunity (SLE). Serum levels have been shown to correlate with disease activity in autoimmune hepatitis Elevated serum levels of TGF-1 are determined in chronic fatigue syndrome patients and in Guillain-Baire syndrome patients . An inverse correlation with disease activity was described for TGF1 levels in Kawasaki disease and patients with IgA deficiency. TGF-1 has been confirmed to promote fibrotic processes, thus it is implicated in the myelofibrosis with myeloid metaplasia. Increased serum levels of TGF-1 in patients affected by thrombotic thromocytopenic purpura implicate its function on bone marrow haematopoiesis . Determination of circulating TGF-1 turned out to reflect the various stages in solid tumors as has been shown for cervical cancer , elevations were furthermore found in prostate cancer , bladder cancer , and liver cancer. Decreased levels of TGF-1 in the serum of sepsis and acute stroke patients may reflect the changing immunological-inflammatory status of these patients. Decreased TGF-1 serum levels were described for patients with acute Plasmodium falciparum malaria . Cytokines CE- Part5- Hematopoiesis-Differentiation June 2008 3/5 3. IL-5/ IL-13/ TGF-1- Technical performances IL-13 ELISA IL-5 ELISA Cat # Label Format KAPMS120 Cat # Biotin-SAV -conjugate KAPMS113 Label Format Biotin-SAV conjugate Standard Format Concentrated (to be diluted) Standard Format Concentrated (to be diluted) Standard Range 500 - 7.8 pg/ml Standard Range 100 - 1.6 pg/ml Sample Size 50 µl Sample Size 50 µl Sample Type Serum, Plasma Sample Type Serum, Plasma Controls Included no Controls Included no Incubation 190 min with shaking Sensitivity 1.45 pg/ml Intra Assay 6.6% Sensitivity 0.73 pg/ml Inter Assay 6.8% Intra Assay 6.0% Inter Assay 4.6% Calibration NIBSC 94/622 135 min with shaking at 100 Incubation rpm TGF-beta1 ELISA Label Format Standard Format Standard Range Sample Size KAPMS105 HRP-conjugate Concentrated (to be diluted) 30 - 0.47 ng/ml 20 µl Serum, Plasma (need to Sample Type be pretreated for 60 min.) Controls Incubation Time Sensitivity no 1,5 ABSORPTION 450 nm Cat. Nr. 1 0,5 255 min w ith shaking 23.8 pg/ml Intra-assay CV 6.7% Inter-assay CV 8.5% 0 0 5 10 15 20 25 TGF-beta1 CONCENTRATION [ng/ml] Cytokines CE- Part5- Hematopoiesis-Differentiation June 2008 4/5 30 Cardiov ascular & Salt Balance 4. Package inserts The package inserts will be available on BioSource website. For more information on those products, please visit our website: www.biosource-diagnostics.com Cytokines CE- Part5- Hematopoiesis-Differentiation June 2008 5/5