IFNG Monkey
Description
Introduction to IFNG Monkey
Interferon-gamma (IFNG) in non-human primates, commonly referred to as "IFNG Monkey," is a type II cytokine critical for immune regulation. Produced by activated T-cells and natural killer (NK) cells, it plays a central role in antimicrobial, antiviral, and antitumor responses by enhancing antigen presentation and macrophage activation . IFNG Monkey shares 94% amino acid sequence identity with human IFNG, enabling cross-reactivity in research applications .
Key Functions
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Immune Activation: Enhances MHC class I/II expression, promotes immunoproteasome assembly, and increases nitric oxide synthase (iNOS) activity .
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Antiproliferative Effects: Inhibits tumor growth by upregulating fibronectin and MHC molecules, improving immunorecognition .
ELISA Kits
Commercial kits are optimized for IFNG detection in serum, plasma, and cell cultures:
| Kit | Sensitivity | Sample Types | Source |
|---|---|---|---|
| Abcam ab270895 | 90-min protocol | CSF, plasma, serum | |
| BD OptEIA™ | 1–1000 pg/mL | NHP serum, plasma, supernatants | |
| MabTech ELISA Pro | 1 pg/mL | Serum, plasma, supernatants |
Interferon-Gamma Release Assays (IGRA)
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Monkey IGRA (mIGRA): Combines human QFT-Plus stimulation tubes with species-specific IFNG detection, achieving 36% sensitivity in tuberculosis (TB) screening .
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Mitogen Optimization: Concanavalin A (Con A) + Pokeweed mitogen (PWM) induces the highest IFNG secretion in macaques (OD 450 = 4.270 vs. 0.258 for PHA alone) .
Tuberculosis Detection
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Diagnostic Sensitivity: In a 12-month study, mIGRA detected TB in 36% of cynomolgus macaques, outperforming tuberculin skin tests (26%) and cultures (21%) .
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Kinetics: IFNG levels peak 5–6 weeks post-MTB infection, then decline as infection is controlled .
Cancer Immunotherapy
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Antitumor Activity: Recombinant IFNG enhances MHC presentation and synergizes with checkpoint inhibitors (e.g., durvalumab) in lung and bladder cancers .
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Limitations: Prolonged IFNG exposure may upregulate PD-L1 in ovarian cancer, promoting immune evasion .
Recombinant IFNG Monkey Proteins
| Product | Source | Purity | Applications |
|---|---|---|---|
| Recombinant Cynomolgus IFNG | Yeast expression | >85% | SDS-PAGE, cell stimulation |
| IFNG Rhesus Macaque | E. coli | >90% | Antiviral assays, bioactivity |
Challenges and Future Directions
Product Specs
Interferon-gamma (IFN-gamma) is a cytokine primarily released by activated lymphocytes, including T cells and natural killer cells, in response to specific antigens or mitogens. Beyond its antiviral properties, IFN-gamma plays a crucial role in regulating immune responses. It acts as a potent activator of macrophages, exhibits antiproliferative effects on transformed cells, and can enhance the antiviral and antitumor actions of type I interferons.
This product consists of recombinant Interferon-gamma (IFNG) from Rhesus Macaque, produced in E.Coli. It is a non-glycosylated polypeptide chain composed of 142 amino acids, with a molecular weight of approximately 16.8kDa. The purification of IFNG is achieved through proprietary chromatographic methods.
This product appears as a sterile, filtered, white lyophilized (freeze-dried) powder.
This product is lyophilized from a 0.2µm filtered concentrated solution in phosphate-buffered saline (PBS) at a pH of 7.4.
To reconstitute the lyophilized Interferon-gamma Rhesus Macaque, it is recommended to dissolve it in sterile 18MΩ-cm H₂O at a concentration not less than 100µg/ml. This solution can be further diluted in other aqueous solutions as needed.
Lyophilized IFNG remains stable at room temperature for up to 3 weeks. However, for long-term storage, it is recommended to store the desiccated product below -18°C. After reconstitution, Interferon-gamma Rhesus Macaque should be stored at 4°C for a period of 2-7 days. For extended storage, it should be kept below -18°C. It's crucial to avoid repeated freeze-thaw cycles to maintain product integrity.
The purity of this product is determined to be greater than 97.0% through the following analyses:
(a) Reverse-phase high-performance liquid chromatography (RP-HPLC).
(b) Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
The half-maximal effective concentration (ED50) of this product, as determined by an antiviral assay using human HeLa cells infected with encephalomyocarditis (EMC) virus, is less than 20.0 ng/ml. This corresponds to a specific activity greater than 5.0 x 10⁴ international units per milligram (IU/mg).
Immune IFN, type II IFN, T cell IFN, MAF, IFNG, IFG, IFI, IFN-gamma.
Escherichia Coli.
QDPYVKEAEN LKKYFNAGDP DVADNGTLFL DILRNWKEES DRKIMQSQIV SFYFKLFKNF KDDQRIQKSV ETIKEDINVK FFNSNKKKRD DFEKLTNYSV TDSNVQRKAV HELIQVMAEL SPAAKIGKRK RSQMFRGRRA SQ.
Q&A
What is the functional role of IFNG in cynomolgus monkey immune responses?
IFNG acts as a pleiotropic cytokine coordinating innate and adaptive immunity through JAK-STAT signaling. In cynomolgus models, it enhances antigen presentation by upregulating immunoproteasome subunits (e.g., PA28) and MHC class I/II complexes, increasing peptide diversity for immune recognition . Experimental validation requires measuring IFNG-induced changes in immune cell populations via flow cytometry paired with transcriptional analysis of STAT1 target genes like IRF1 . Baseline IFNG levels in healthy monkeys show significant biological variation (Table 1):
| Sample Type | Mean Concentration (pg/mL) | Inter-Assay CV% | Source |
|---|---|---|---|
| Serum | 148.3 ± 22.1 | <10% | Assay Genie |
| Cell Culture Supernatant | 89.7 ± 14.6 | <8% | Abcam |
How should researchers design experiments measuring IFNG in monkey models?
Optimal experimental design requires:
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Sample size justification: Power analysis based on expected effect sizes. For IFNG knockout studies, n=6/group detects 2-fold changes (α=0.05, β=0.8) given observed biological variation .
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Longitudinal sampling: Plasma IFNG levels exhibit diurnal fluctuations (peak: 08:00–12:00; trough: 20:00–24:00) requiring standardized collection times .
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Assay validation: Cross-reactivity testing against homologs (94% identity to human IFNG ) and spike-recovery validation in target matrices (Table 2):
| Matrix | Spiked Concentration (pg/mL) | Recovery Rate (%) |
|---|---|---|
| Cerebral Spinal Fluid | 250 | 92–108 |
| Tissue Homogenate | 500 | 85–97 |
How to resolve discrepancies between IFNG detection methods in primate studies?
Conflicting results often arise from:
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Assay sensitivity thresholds: Low-abundance IFNG in latent infections (31.25–2000 pg/mL detectable range ) may be missed by less sensitive platforms.
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Temporal expression dynamics: RNA-seq identifies IFNG-responsive genes (e.g., CXCL9/10) peaking at 6–8h post-stimulation, while protein detection lags by 12–24h .
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Diagnostic discordance: In tuberculosis studies, 37.5% of culture-positive monkeys tested negative via tuberculin skin test but were detected by IFNG-release assays (mIGRA) .
Resolution strategy:
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Use orthogonal methods (e.g., ELISA + RNAscope in situ hybridization ).
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Apply mixed-effects models to account for inter-individual variability in longitudinal studies.
What are best practices for engineering recombinant IFNG in monkey cell lines?
The baculovirus expression system achieves high-yield IFNG production (10^8 units/mL ):
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Clone IFNG cDNA via RT-PCR using primers spanning the 165-aa coding sequence .
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Optimize secretion using insect cell glycosylation signals (e.g., gp67 leader sequence).
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Validate bioactivity via antiviral assays using VSV-GFP reporter systems (EC50: 0.1–1.0 ng/mL ).
Critical parameters:
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Post-translational modifications: Monkey IFNG has 2 N-glycosylation sites absent in murine homologs .
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Stability: Lyophilized standards maintain activity for 6 months at -80°C .
How does IFNG influence mesenchymal stem cell (MSC) immunomodulation in transplantation models?
RNA-seq of IFNG-treated cynomolgus MSCs reveals:
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Upregulation of IDO1 (15.4-fold) and PD-L1 (9.2-fold) mediating T-cell suppression .
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Downregulation of osteogenic markers (RUNX2: -3.1-fold) indicating lineage-specific effects.
Mitigation approaches:
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Include heparin/EDTA controls in sample processing.
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Use single-cell RNA-seq to disentangle cell-type-specific IFNG responses.
Product Science Overview
Interferon-gamma (IFN-γ) is a critical cytokine in the immune system, known for its role in modulating immune responses and exerting antiviral, immunoregulatory, and anti-tumor properties. Recombinant Interferon-gamma derived from Rhesus Macaque (Macaca mulatta) is a valuable tool in biomedical research, particularly in studies involving non-human primate models.
Interferon-gamma is also referred to as type II or immune interferon. It is a dimeric protein composed of two identical subunits, each with a molecular weight of approximately 20-25 kDa. The protein is variably glycosylated, which can influence its stability and activity . IFN-γ is produced primarily by T lymphocytes and natural killer (NK) cells in response to antigenic or mitogenic stimuli.
The primary function of IFN-γ is to activate macrophages, enhance antigen presentation, and promote the differentiation of T helper cells into Th1 cells. It also plays a crucial role in the regulation of immune responses against intracellular pathogens, such as viruses and certain bacteria .
Recombinant Rhesus Macaque IFN-γ is produced using E. coli expression systems. The recombinant protein is typically purified to a high degree of purity (>97%) and is available in both carrier-free and carrier-containing formulations. The carrier protein, often Bovine Serum Albumin (BSA), is added to enhance protein stability and shelf-life .
Recombinant Rhesus Macaque IFN-γ is widely used in various research applications, including:
- Immunological Studies: It is used to study the immune responses in non-human primate models, particularly in the context of infectious diseases and vaccine development.
- Antiviral Assays: The antiviral activity of IFN-γ can be assessed using cell-based assays, such as those involving HeLa cells infected with encephalomyocarditis virus .
- Inflammatory Response Studies: IFN-γ is a key player in inflammatory responses, and its role can be studied in models of diseases such as COVID-19. For instance, in rhesus macaques, IFN-γ has been shown to drive the development of pulmonary lesions during SARS-CoV-2 infection .
