Very Early-Onset Inflammatory Bowel Disease (VEO-IBD) refers to inflammatory bowel disease with onset before 6 years of age, characterized by recurrent colitis, perianal lesions, and malabsorption as the main clinical manifestations, accounting for approximately 15% of pediatric IBD patients. Compared to IBD in adults and older children, VEO-IBD exhibits significant differences in clinical presentation, severity, complications, and treatment response, with genetic factors playing a crucial role in its pathogenesis. Currently, VEO-IBD has been associated with approximately 70 monogenic defects, among which mutations in IL-10 and IL-10 receptor A/B genes have been widely reported. The TR-FRET IL-10/IL-10RA assay kit can be used to quantitatively measure the binding activity of IL-10 to its receptor, providing a technical tool for studying the molecular mechanisms of defects in this signaling pathway.
Defects in the IL-10 signaling pathway include mutations in the coding regions of the IL-10 gene and IL-10 receptor genes, leading to dysregulated anti-inflammatory responses and severe enterocolitis. The IL-10 receptor consists of two subunits: IL-10RA and IL-10RB. In East Asian populations, IL-10RA mutations are more common than IL-10 and IL-10RB mutations, accounting for approximately 10% of VEO-IBD cases. IL-10-deficient patients typically develop symptoms within the first 3 months of life, often presenting with colitis and perianal lesions, accompanied by severe bloody diarrhea. Patients with IL-10 receptor defects usually develop refractory diarrhea or recurrent oral ulcers within the first year of life, with a high frequency of extraintestinal manifestations, including dermatological features and growth retardation. TR-FRET technology can be applied to assess changes in IL-10 binding capacity to mutant receptors and evaluate the impact of different mutations on receptor-ligand interactions.
IL-10 is a protein encoded by the IL-10 gene located on the long arm of chromosome 1. It is a 36 kDa homodimer composed of two polypeptide chains, each containing 160 amino acids. IL-10 is secreted by both myeloid and lymphoid cells in response to various stimuli, including T and B lymphocytes, monocytes, macrophages, natural killer cells, neutrophils, mast cells, dendritic cells, and epithelial cells. As a critical anti-inflammatory cytokine in humans, IL-10 regulates innate and adaptive immunity by influencing the expression of pro-inflammatory molecules and the functions of various immune cells. IL-10 inhibits dendritic cell maturation and antigen-presenting functions, suppresses Toll-like receptor-mediated MyD88-dependent pathways, and helps downregulate the expression of inflammatory cytokines such as IL-6, IL-1β, and TNF. These processes are essential for maintaining gastrointestinal immune homeostasis. The TR-FRET IL-10/IL-10RA assay kit can be used to study the signaling activation process following IL-10-receptor binding and evaluate signal transduction efficiency under different conditions.
Upon binding to the IL-10 receptor, IL-10 activates JAK1 and TYK2, leading to phosphorylation of IL-10RA and subsequent phosphorylation of STAT3. This results in STAT3 dimerization, nuclear translocation, and promotion of target gene transcription, ultimately expressing anti-inflammatory effectors through downstream signaling cascades. IL-10 mutant mice spontaneously develop IBD, characterized by a surge in inflammatory cytokines. Mutant IL-10 receptors fail to transduce IL-10-induced anti-inflammatory signals, resulting in insufficient STAT3 phosphorylation and increased secretion of TNF and other pro-inflammatory cytokines by peripheral blood mononuclear cells. In IL-10RA-mutant B cells, reduced IL-10-induced SOCS3 expression renders B cells unresponsive to negative feedback regulation by IL-10, leading to hyperinflammatory immune responses in the gut. TR-FRET technology can be applied to screen molecules capable of restoring IL-10 binding to mutant receptors, providing quantitative analytical tools for developing targeted therapeutic strategies.
Allogeneic hematopoietic stem cell transplantation is currently the only curative treatment for VEO-IBD caused by IL-10 signaling pathway defects, but it carries significant risks and requires careful patient selection. Pharmacological treatments primarily include 5-aminosalicylic acid, immunomodulators, and biologics such as anti-TNF antibodies, but their efficacy is suboptimal. IL-10 supplementation therapy has demonstrated good safety and tolerability in clinical trials, but its effectiveness in inducing Crohn's disease remission remains limited, possibly due to factors such as low drug stability and poor bioavailability. Extracellular vesicle-based mRNA therapies have shown anti-inflammatory potential in IBD, offering a promising new direction for future treatments. Anti-IL-1 therapies like anakinra have proven effective in some IL-10 receptor-deficient patients and may serve as a bridge to hematopoietic stem cell transplantation. Fecal microbiota transplantation has shown benefits in short-term treatment of active ulcerative colitis for inducing clinical and endoscopic remission.
VEO-IBD caused by IL-10 signaling pathway defects is characterized by early onset, severe clinical manifestations, and poor response to standard therapies. Enhanced clinical awareness of this condition facilitates early identification and timely intervention. The TR-FRET IL-10/IL-10RA assay kit provides a sensitive and efficient detection tool for studying the molecular mechanisms of IL-10 signaling pathway defects, enabling quantitative analysis of receptor-ligand binding activity and drug screening.
The "UniOne® TR-FRET Human IL10/IL10RA Binding Kit" (Catalog No.: UA086016), independently developed by Nanjing UA-Bio Technology Co., Ltd. (UA-Bio), is a high-performance analytical platform specifically designed for studying the interaction between interleukin-10 (IL-10) and its specific receptor IL-10Rα. Based on time-resolved fluorescence resonance energy transfer (TR-FRET) technology, this kit enables precise and efficient evaluation of the binding activity between human IL-10 and IL-10Rα, providing a stable and reliable standardized solution for research in anti-inflammatory immune regulation, autoimmune disease mechanisms, and immunotherapy development.
| Core Advantages | Detailed Parameters / Functional Description |
|---|---|
| High Purity and Intact Biological Activity | The kit's core components include highly purified, biologically active human IL-10 protein (maintaining native homodimeric conformation) and IL-10Rα, both validated through multidimensional quality control. These components retain correct spatial conformation and intact binding functionality, accurately mimicking the high-affinity binding of IL-10 to IL-10Rα under physiological conditions, ensuring experimental accuracy, reproducibility, and functional relevance. |
| Excellent Lot-to-Lot Consistency and Stability | Leveraging an internationally advanced protein expression platform and highly standardized production processes, combined with a stringent quality control system, the product demonstrates exceptional long-term stability and outstanding lot-to-lot consistency, providing reliable quality assurance for long-term, continuous drug screening and mechanistic research. |
| Ready-to-Use Flexible Experimental Platform | This kit employs a homogeneous TR-FRET technology with a simple "add-mix-read" workflow, eliminating the need for cumbersome washing steps. Its optimized formulation is compatible with automated multi-well plate (96/384-well) platforms, making it suitable for various research applications, including anti-IL-10 antibody/antagonist screening, receptor blocker evaluation, competitive binding assays, affinity analysis, and biosimilar activity assessment. |
| Comprehensive Solutions and Professional Support | We provide fully validated standard protocols, typical dose-response curves, and detailed result interpretation guidelines to help establish stable and reproducible experimental workflows. Nanjing UA-Bio's professional technical team offers end-to-end support for research design, experimental optimization, and data analysis. |
Nanjing UA-Bio Technology Co., Ltd. is committed to providing cutting-edge, high-quality core reagents and tools for immunology, cell therapy, and innovative drug development. For detailed technical parameters, validation data, or specific application inquiries regarding the "UniOne® TR-FRET Human IL10/IL10RA Binding Kit" (Catalog No.: UA086016), please feel free to contact us.
