Recombinant Mouse Interleukin-10 protein (Il10) (Active)
Description
Molecular Structure and Biochemical Properties
Recombinant Mouse Interleukin-10 protein is a major immune regulatory cytokine with profound anti-inflammatory functions that limit excessive tissue disruption caused by inflammation . The mature form of mouse IL-10 consists of 160 amino acid residues, with the expression region typically spanning amino acids 19-178 of the full protein sequence . The amino acid sequence includes:
SRGQYSREDNNCTHFPVGQSHMLLELRTAFSQVKTFFQTKDQLDNILLTDSLMQDFKGYLGCQALSEMIQFYLVEVMPQAEKHGPEIKEHLNSLGEKLKTLRMRLRRCHSRFLPCENKSKAVEQVKSDFNKLQDQGVYKAMNEFDIFINCIEAYMMIKMKS
The protein has a theoretical molecular weight of approximately 18.7 kDa, though slight variations exist between different commercial preparations, with reports ranging from 18.6 to 18.8 kDa . Biologically active IL-10 exists as a homodimer, which is critical for its function but presents challenges for therapeutic applications due to its inherent instability and short half-life in vivo .
| Property | Specification |
|---|---|
| Molecular Weight | 18.7-18.8 kDa |
| Amino Acid Length | 160 residues (mature protein) |
| Expression Region | Amino acids 19-178 |
| Active Form | Homodimer |
| Isoelectric Point | Not commonly reported |
| Stability | Unstable in natural form; requires specific storage conditions |
Expression Systems and Production Methods
Recombinant mouse IL-10 is produced using various expression systems, with significant differences in quality and characteristics based on the host system. The two primary expression platforms include:
HEK 293 Expression System
Recombinant mouse IL-10 expressed in Human Embryonic Kidney 293 cells offers advantages in terms of post-translational modifications that more closely resemble the native protein . Products from this system typically achieve:
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Purity levels of ≥95% as determined by SDS-PAGE
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Endotoxin levels of ≤0.005 EU/μg
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Suitability for applications including SDS-PAGE, functional studies, mass spectrometry, HPLC, and cell culture
coli Expression System
E. coli-expressed recombinant mouse IL-10 offers advantages in terms of higher yield and cost-effectiveness, with typical specifications including:
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Purity levels >97% as determined by SDS-PAGE
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Endotoxin levels <1.0 EU/μg measured by LAL method
The production process generally involves multiple chromatography steps followed by sterile filtration through a 0.22 μm filter for preparations intended for cell culture or in vivo use. The final product is supplied in various forms, including frozen liquid comprised of sterile-filtered aqueous buffered solution with glycerol and bovine serum albumin, or as lyophilized powder requiring reconstitution .
Biological Function and Signaling Mechanisms
Interleukin-10, originally known as Cytokine Synthesis Inhibitory Factor (CSIF), shares over a 80% sequence homology with the Epstein-Barr Virus protein BCRFI . The protein is produced by a diverse range of cells including activated Th2 cells, fetal thymocytes, monocytes/macrophages, keratinocytes, B cells, and glial cells .
Signaling Pathway
The mechanistic pathway of IL-10 involves a complex cascade of molecular interactions:
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IL-10 binds to its heterotetrameric receptor comprising IL-10RA and IL-10RB
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This binding leads to activation of Janus kinase 1 (JAK1) and Tyrosine kinase 2 (Tyk2)
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JAK1 and STAT2 mediate phosphorylation of STAT3
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Phosphorylated STAT3 translocates to the nucleus
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Nuclear STAT3 drives expression of anti-inflammatory mediators
Recent research has re-evaluated the roles of IL-10R2 and IL-10R2-associated signaling via Tyk2, revealing that IL-10-mediated responses depend on both IL-10R1 and IL-10R2 across multiple cell types, including bone marrow-derived dendritic cells, macrophages, and mast cells .
Immunoregulatory Effects
Recombinant mouse IL-10 exhibits multiple interrelated biological activities:
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Inhibition of macrophage-mediated cytokine synthesis
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Suppression of the delayed-type hypersensitivity response
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Stimulation of the Th2 cell response resulting in elevated antibody production
IL-10 primarily targets antigen-presenting cells such as macrophages and monocytes, inhibiting their release of pro-inflammatory cytokines including GM-CSF, G-CSF, IL-1α, IL-1β, IL-6, IL-8, and TNF-α . Additionally, it interferes with antigen presentation by reducing expression of MHC-class II and co-stimulatory molecules, thereby inhibiting T cell activation .
The protein also enhances B cell survival, proliferation, and antibody production while blocking NF-kappa B activity . The net effect appears to be inhibition of pro-inflammatory T cell-mediated immunity .
Research Applications
Recombinant mouse IL-10 serves numerous research purposes across immunology and inflammation studies:
In Vitro Applications
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Investigation of cytokine inhibition mechanisms
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Studies on immune cell differentiation and function
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Analysis of anti-inflammatory signaling pathways
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Standards for enzyme-linked immunosorbent assays (ELISAs)
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Cell culture supplements for generating regulatory T cells or anti-inflammatory macrophages
In Vivo Applications
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Animal models of autoimmune diseases
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Studies of inflammatory bowel disease
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Investigation of allergic conditions
Functional Assays
Recombinant mouse IL-10 is commonly used to assess biological activity through:
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Inhibition of cytokine production in LPS-stimulated macrophages
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JAK-STAT signaling pathway activation
Engineered IL-10 Variants and Recent Advances
A significant limitation of using IL-10 in clinical applications is that the biologically active form is an unstable homodimer with a short half-life that degrades easily in vivo . Consequently, IL-10 therapy using recombinant native IL-10 has shown limited success in treating human disease.
Novel Stable IL-10 Dimer
To improve therapeutic potential, researchers have developed a novel form of IL-10 consisting of two IL-10 monomer subunits linked in a head-to-tail fashion by a flexible linker . This engineered protein demonstrates:
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Improved temperature and pH-dependent biological stability
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Greater activity than natural IL-10 both in vitro and in vivo
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Similar binding to the IL-10 receptor as natural IL-10
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More effective suppression of LPS-induced inflammation in vivo
Interestingly, studies showed that the linker length itself did not affect the expression and biological activity of this stable IL-10 molecule, offering flexibility in design for future therapeutic applications .
IL-10 Reporter Mouse Models
Another area of active research involves the development of IL-10 reporter mouse strains for identifying IL-10-producing cells in vivo. To date, at least eight IL-10 reporter strains have been published, reflecting the importance and challenges in tracking IL-10-producing cells in various disease contexts .
Researchers have proposed developing "multi-cytokine" reporter mouse models that would enable highly sensitive detection of multiple cytokines within the same cell, potentially revolutionizing our understanding of cytokine-producing cells in health and disease .
Disease Associations and Therapeutic Potential
Knockout studies in mice have suggested that IL-10 is an essential immunoregulator in the intestinal tract . Mutations in the IL-10 gene have been associated with increased susceptibility to several conditions:
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HIV-1 infection
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Rheumatoid arthritis
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Graft-versus-host disease
These disease associations highlight the potential therapeutic value of recombinant IL-10 or engineered IL-10 variants in treating inflammatory and autoimmune conditions. The development of stable IL-10 dimers with improved pharmacokinetic properties represents a significant step toward realizing this therapeutic potential .
Product Specs
Target Background
- G protein-coupled receptor 39 exhibits an anti-inflammatory activity by enhancing IL-10 production from macrophages PMID: 30053407
- Our findings provide direct evidence that the capacity of NK cells to secrete IL-10 is required to maintain the successful "innate" modulation of DC phenotype in the gravid uterus. PMID: 28526846
- Restrictive IL-10 induction by an innocuous parainfluenza virus vector ameliorates nasal allergy. PMID: 27555458
- A regulatory loop exists where IL-10 directly restricts CD8(+) T cell activation and function through modification of cell surface glycosylation, enabling the establishment of chronic infection. PMID: 29396160
- Genome-wide knockdown of 19 ribosomal proteins resulted in decreased IL-10 and increased TNF-alpha production. PMID: 29657255
- YB-1 orchestrates onset and resolution of renal inflammation via IL-10 gene regulation. PMID: 28664613
- This study demonstrates IL-10 production and T cell-suppressive capacity in B cell subsets from atherosclerotic apoE (-/-) mice PMID: 28744806
- IL-10 Knock out-Endothelial progenitor cell-Exosomes were highly enriched in microRNAs and proteins that promote inflammation and apoptosis and inhibit angiogenesis. PMID: 28471299
- We demonstrated the IL-10 expression of the pyramidal neurons in CA2 and CA3 subregions, for the first time globally, after infection with the Mu-3 virus PMID: 28493345
- These data indicate that CD4(+) follicular regulatory T (Tfr) cells play a multifaceted role in the fine-tuning of the germinal center response and identify IL-10 as an important mediator by which Tfr cells support the germinal center reaction PMID: 29054998
- Decreased interleukin-10 (IL-10) expression was found in the hippocampus of the stressed mice, while no differences in pro-inflammatory cytokine expression and tryptophan (TRYP), kynurenine (KYN) or 3-hydroxy kynurenine (3-HK) levels were found. PMID: 28789949
- HBV-specific CD8(+) T cells produce IL-10 upon antigen recognition and that this cytokine enhances CD8(+) T cell survival. PMID: 28483675
- Novel regulatory T-cells that are induced by B cells and do not express Foxp3 and IL-10 alleviate intestinal inflammation in vivo. PMID: 27581189
- Results provide evidence that macrophage IL-10 production is regulated by NLRP3 and contributes to the pathophysiology of doxorubicin-induced cardiotoxicity independently of IL-1beta. PMID: 27225830
- Activation of TLR2, TLR4, and TLR9 induced the production of IL-10 in microglia to a greater extent than activation of TLR3. Combination of TLR3 triggering with the other TLRs, enhanced IL-10 through the modulation of its transcription, via interferon (IFN)-beta, but independently of IL-27. Presence of IFN-gamma in the microenvironment abrogated the modulation of IL-10 by TLR3, whereas that of IL-17 had no effect. PMID: 28617991
- The absence of IL-10 led to longer illness, more weight loss, more death, and slower viral clearance than in mice that produced IL-10. IL-10 influenced development of disease-causing T cells and entry into the brain of B cells producing antiviral antibody. PMID: 29263262
- Cytokine-inducing and anti-inflammatory activity of chitosan and its low-molecular derivative. PMID: 29513410
- TonEBP suppresses M2 phenotype via downregulation of the IL-10 in M1 macrophages. PMID: 27160066
- Nod2-signaling is essentially involved in the well-balanced innate and adaptive immune responses upon Campylobacter jejuni infection of IL-10(-/-) mice. PMID: 28752081
- CD8(+) T cell-derived IL-10 does not contribute significantly to the resolution of contact hypersensitivity responses. PMID: 27714845
- Docosahexaenoic acid activates GPR120 to prevent experimental colitis in IL-10 deficient mice. PMID: 28039475
- In a Leptospira-infected mouse model, the study showed evidence of a possible role of IL-10 on host susceptibility, bacterial clearance, and on regulation of cytokine gene expression PMID: 28410507
- In vivo IL-10 treatment increased fibroblast activation (proliferation, migration, and collagen production), an effect that was both directly and indirectly influenced by macrophage M2 polarization PMID: 28439731
- Suggest that the IL-17A/IL-10/STAT3 signaling pathway plays a crucial role in the pathogenesis of hepatic fibrosis by suppressing hepatocellular autophagy and that blocking this pathway may provide therapeutic benefits for the treatment of hepatic fibrosis. PMID: 28039485
- Mechanistic studies suggest a PKC-Syk-mediated signaling pathway, to which IL-10 conversely inhibits, is required for activating macrophage self-targeting, followed by phagocytosis independent of calreticulin Moreover, we identified spleen red pulp to be one specific tissue that provides stimuli constantly activating macrophage phagocytosis albeit lacking in Cd47(-/-) or Sirpalpha(-/-) mice. PMID: 27578867
- High IL10 expression is associated with lung cancer. PMID: 26956044
- The activation of STAT3 was much higher in Gal12(-/-) macrophages activated by lipopolysaccharide, which was correlated with higher levels of IL-10. Adipocytes showed higher insulin sensitivity when treated with Gal12(-/-) macrophage-conditioned media than those treated with Gal12(+/+) macrophages. PMID: 26873172
- This study reveals a key role of IL-10 in controlling cellular metabolism via inhibiting mTORC1, and this metabolic control by IL-10 is critical to control of inflammation. PMID: 28473584
- IL-10-MSCs offered superior protection against LPS-induced ALI. PMID: 29072959
- Methane-rich saline may activate the PI3K-AKT-GSK-3beta pathway to induce IL-10 expression and produce anti-inflammatory effects via the NF-kappaB and MAPK pathways. The findings provide a new pharmacological strategy for management of inflammatory response after acute liver injury. PMID: 28597201
- Plasma adiponectin and leptin were also decreased in IL 10tm. These findings suggest that frailty observed in this mouse model of chronic inflammation may in part be driven by alterations in fat mass, hormone secretion, and energy metabolism PMID: 29267271
- We believe that current findings derived from human and mouse experiments will promote the development of new drugs and therapies based on IL-10 modulation, which may enhance host immunity and bacterial clearance during infection. However, because IL-10 can play both favorable and unfavorable effects over the host depending on the bacterial infection, it may act as a double-edged sword. PMID: 27522641
- IL-12p35 suppresses lymphocyte proliferation, induces expansion of IL-10-expressing and IL-35-expressing B cells and ameliorates autoimmune uveitis. PMID: 28959012
- Aerobic interval training enhanced the anti-inflammatory indices IL-10/TNF-alpha ratio and IL-15 expression in skeletal muscle in tumor-bearing mice. PMID: 27863332
- Both IL-6 protein production and transcript levels were downregulated by RA in respiratory tract epithelial cells (LETs), but upregulated in macrophages (MACs). RA also increased transcript levels of MCP-1, GMCSF, and IL-10 in MACs, but not in LETs. Conversely, when LETs, but not MACs, were exposed to RA PMID: 27940088
- DnaK was able to induce TGF-beta mRNA in treated macrophages in an IL-10 dependent manner. PMID: 27337694
- Findings suggest that the axis IL-10/claudin-10 is a promising target for the development of therapeutic agents against aggressive melanoma PMID: 29145406
- IL-10 signaling in CD11c+ cells controls small intestinal immune homeostasis by limiting reactivation of local memory T cells and to protect against Helicobacter hepaticus-induced colitis. PMID: 27027442
- Imperatorin exerts antiallergic effects in Th2-mediated allergic asthma via induction of IL-10-producing regulatory T cells by modulating the function of dendritic cells. PMID: 27185659
- Cisplatin induces immune-suppressive tolerogenic dendritic cells in TLR agonist-induced inflammatory conditions via abundant IL-10 production, thereby skewing Th cell differentiation towards Th2 and Tr1 cells. This relationship may provide cancer cells with an opportunity to evade the immune system. PMID: 27172902
- Following vasectomy, IL1alpha, IL1beta, IL1ra, IL10, and TNF-alpha may mediate immune reaction in whole epididymis, whereas IL6 and TGF-beta1 may mediate regionally different immune response primarily in the lower part of epididymis. PMID: 27476761
- This study shows that progesterone and estradiol inhibit the production of IL-10 by activated B cells PMID: 27317920
- Il-10 deficient mice express IFN-gamma mRNA and clear Leptospira interrogans from their kidneys more rapidly than normal C57BL/6 mice PMID: 28237664
- Depletion of Tregs increased adaptive T cell responses and deficiency of IL10 reduced morbidity and conferred enhanced protection against influenza virus. PMID: 28086957
- Data show that the severity of experimental autoimmune encephalomyelitis (EAE) were reduced and the serum IL-10 expression levels were increased in CD226 knockout mice than that in control mice when both received EAE induction. PMID: 26942885
- This study shows that Influenza A virus-induced release of IL-10 inhibits the anti-microbial activities of invariant natural killer T cells during invasive pneumococcal superinfection PMID: 27220813
- Data show that two functionally distinct cytokines, interleukin-4 (IL-4) and interferon-gamma (IFN-gamma), significantly potentiate the ability of mesenchymal stem cells (MSCs) to inhibit interleukin-10 (IL-10) production by activated regulatory B cells (Bregs). PMID: 27665290
- The anti-inflammatory functions of p38 MAPK in macrophages are critically dependent on production of IL-10. PMID: 28877953
- Pretreating mice with carrageenan once a day before injecting LPS increased the levels of IL-10 by 2.5-fold and reduced TNF-alpha production by 2-fold compared with control. So, kappa/beta-carrageenan alone and in combination with LPS enhanced the cellular activity and mobility of peritoneal macrophages by increasing cell adhesion and migration compared with control. PMID: 28130856
- Results demonstrate that IL-10-producing interstitial macrophages negatively regulate Th2- and Th17-mediated inflammatory responses, helping prevent neutrophilic asthma PMID: 26976823
