IL 2 Equine

Interleukin-2 Equine Recombinant
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Description

Biological Functions and Mechanisms

IL-2 Equine regulates immune responses by binding to heterotrimeric receptors (CD25/CD122/CD132) on lymphocytes, promoting:

  • T-cell proliferation and differentiation into effector/memory subsets .

  • B-cell growth in horses, unlike IL-4, which lacks this activity .

  • Natural killer (NK) cell activation, enhancing antitumor responses .

  • Regulatory T-cell (Treg) expansion, preventing autoimmune reactions .

In equine peripheral blood mononuclear cells (PBMCs), IL-2 induces dose-dependent proliferation (ED50ED_{50}: 0.3–1.5 µg/mL) .

Production and Purification Methods

IL-2 Equine is produced via recombinant DNA technology:

Expression SystemKey Features
E. coliNon-glycosylated, cost-effective, high yield .
Mammalian CHO cellsPost-translational modifications (e.g., glycosylation for IL-4) .
YeastEndotoxin-free, >98% purity, suitable for cell culture and diagnostics .

Purification: Proprietary chromatographic techniques or His-tag affinity chromatography .

Sarcoid Tumor Treatment

IL-2 Equine has been tested in clinical trials for equine sarcoids, a common skin tumor:

StudyProtocolEfficacyAdverse Effects
ALVAC-fIL2 (canarypox vector) Four intratumoral injections over 7 weeks86% tumor reduction; median time to response: 89 daysTransient inflammation in 2/14 horses
Recombinant canarypox-fIL2 Two injections (day 0, day 7)50% complete/partial regressionMild fever in 1/20 horses
Cisplatin + IL-2 Local cisplatin + single high-dose IL-253% complete remissionNot reported

Limitations

  • Low-dose IL-2 alone showed only 14% efficacy .

  • Species specificity: Equine IL-2 does not stimulate murine CTLL-2 cells .

Research Findings and Clinical Insights

  • Dual Role in Immunity: IL-2 Equine enhances antitumor responses while suppressing autoimmunity via Tregs .

  • Glycosylation Variants: Equine IL-4 exhibits three glycosylated forms (17.1–22.1 kDa), unlike IL-2 .

  • Stability Enhancements: Adding 0.1% HSA/BSA extends shelf life and prevents aggregation .

Future Directions

  • Optimized Delivery: Viral vectors (e.g., ALVAC) may improve localized IL-2 delivery .

  • Combination Therapies: Synergy with cisplatin or dendritic cell vaccines warrants further study .

Product Specs

Introduction
Interleukin 2 (IL-2) is a cytokine that plays a crucial role in the proliferation of T and B lymphocytes, which are essential components of the immune system. The receptor for IL-2 is a complex structure on the surface of these cells, and its gamma chain is also used by other interleukins, IL-4 and IL-7. Interestingly, the expression of the IL-2 gene in mature thymocytes (T cell precursors) is monoallelic, meaning only one copy of the gene is active. This unusual regulatory mechanism highlights the importance of precise IL-2 expression. Studies in mice have shown that disrupting the IL-2 gene leads to inflammatory bowel disease resembling ulcerative colitis, underscoring its vital role in a healthy immune response to foreign substances.
Description
Recombinant Equine Interleukin-2, produced in E.Coli, is a single-chain polypeptide that is not glycosylated. It consists of 129 amino acids, with a cysteine residue at position 141 replaced by serine compared to the naturally occurring form. This recombinant protein has a molecular weight of 14.9 kDa and is purified using proprietary chromatographic techniques.
Physical Appearance
White, lyophilized powder that has been sterile filtered.
Formulation
Lyophilized from a concentrated solution in phosphate-buffered saline (PBS) at pH 6.0, filtered through a 0.2 µm filter.
Solubility
To reconstitute the lyophilized IL-2, it is recommended to dissolve it in sterile, 18 MΩ-cm water to a concentration of at least 100 µg/ml. This solution can then be further diluted in other aqueous solutions as needed.
Stability
Lyophilized Interleukin-2, while stable at room temperature for up to 3 weeks, should ideally be stored in a dry environment below -18°C. Once reconstituted, IL-2 should be stored at 4°C for short-term use (2-7 days) and frozen below -18°C for long-term storage. For extended storage, adding a carrier protein such as 0.1% HSA or BSA is recommended. Avoid repeated freeze-thaw cycles to maintain protein integrity.
Purity
The purity of this product is greater than 97.0%, as determined by SDS-PAGE and HPLC analysis.
Biological Activity
The ED50, which represents the concentration at which the protein exhibits half of its maximal biological activity, is determined by measuring the proliferation of murine CTLL-2 cells. In this assay, the ED50 is less than 1.0 µg/ml, corresponding to a specific activity greater than 1000 IU/mg.
Synonyms
Interleukin-2, IL-2, T-cell growth factor, TCGF, IL2.
Source
Escherichia Coli.
Amino Acid Sequence
APTSSSKRET QQQLKQLQMD LKLLLEGVNN NKNPKLSKML TFKINMPKKA TELKHLQCLE EELKPLEEML KNFLSKDIKE LMSNINVTVL GLKGSETRFT CEYDDETGTI VEFLNKWITF SQSIFSTMT.

Product Science Overview

Structure and Production

Recombinant Equine Interleukin-2 (rEqIL-2) is produced in Escherichia coli (E. coli) as a single, non-glycosylated polypeptide chain containing 129 amino acids. The recombinant form often includes a substitution of serine for cysteine at position 141 (Cys141Ser), resulting in a molecular mass of approximately 14.9 kDa . The protein is purified using proprietary chromatographic techniques to ensure high purity and biological activity .

Biological Activity

IL-2/IL-2R signaling is essential for T cell proliferation and other fundamental immune functions. The receptor for IL-2 consists of three subunits: IL-2Rα (55 kDa), IL-2Rβ (75 kDa), and the common gamma chain (γc/IL-2Rγ, 64 kDa). These subunits are present on the cell surface in varying preformed complexes .

Recombinant equine IL-2 has been shown to augment proliferation in equine peripheral blood mononuclear cells (PBMCs) and enhance cellular proliferation. It significantly upregulates the transcription and secretion of various cytokines, including IFN-γ, TNF-α, IL-4, IL-10, and IL-12p35 .

Applications

Recombinant equine IL-2 is used in various research and clinical applications, particularly in immunology and veterinary medicine. It is utilized to study the immune response in horses and to develop treatments for equine diseases. The recombinant form allows for controlled studies and consistent results, making it a valuable tool in scientific research .

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