GO:0035395 negative regulation of chemokine (C-X-C motif) ligand 9 production: Immune Regulation, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0035395 describes any process that stops, prevents, or reduces the production of CXCL9 (also known as MIG), a chemokine that recruits CXCR3+ T cells and NK cells.
• CXCL9 production is suppressed by type-I interferons in microglia, revealing a negative feedback mechanism that limits lymphoid chemokine activity in the central nervous system.
• In mesenchymal stem cells, inhibiting CXCL9 expression promotes angiogenesis, linking negative regulation of CXCL9 to vascular remodeling.
• The long non-coding RNA LINC00152 binds EZH2 to regulate the CXCL9/CXCL10/CXCR3 axis and CD8+ T-cell infiltration in gastric cancer, providing a mechanism for negative regulation at the transcriptional level.
• Keratinocyte autophagy supports wound healing by modulating chemokine production, including CXCL9, highlighting a role for autophagy in negative regulation of CXCL9 during tissue repair.
• Understanding GO:0035395 is critical for cancer immunotherapy, autoimmune diseases, and inflammatory conditions where CXCL9 levels dictate immune cell recruitment and disease outcomes [5,8].
Description
Chemokine (C-X-C motif) ligand 9 (CXCL9), also known as MIG (monokine induced by gamma interferon), is a small cytokine that plays a pivotal role in the recruitment of CXCR3-expressing T cells, NK cells, and other immune cells to sites of inflammation and tumors. The production of CXCL9 is tightly controlled at multiple levels, and the Gene Ontology term GO:0035395, negative regulation of chemokine (C-X-C motif) ligand 9 production, encompasses any process that stops, prevents, or reduces the frequency, rate, or extent of CXCL9 production. This regulatory process is essential for preventing excessive immune cell infiltration and tissue damage, and its dysregulation is implicated in cancer, autoimmunity, and chronic inflammatory diseases [4,8]. Research into GO:0035395 has revealed diverse molecular mechanisms, including transcriptional repression by type-I interferons in microglia, regulation by long non-coding RNAs such as LINC00152 in gastric cancer, and modulation by autophagy in keratinocytes during wound healing. These findings underscore the importance of negative regulation of CXCL9 in maintaining immune homeostasis and suggest that targeting this process could have therapeutic benefits in conditions characterized by aberrant CXCL9 expression [5,6]. For researchers, understanding GO:0035395 provides a framework to investigate how cells limit CXCL9 production and how this regulation impacts disease progression. This article synthesizes current knowledge from authoritative QuickGO data and verified PubMed literature to offer a comprehensive overview of the genes, mechanisms, and experimental models relevant to negative regulation of CXCL9 production.
negative regulation of chemokine (C-X-C motif) ligand 9 production At A Glance
| GO ID | GO:0035395 |
|---|---|
| GO term | negative regulation of chemokine (C-X-C motif) ligand 9 production |
| Ontology | biological_process |
| Synonym | negative regulation of CXCL9 production; negative regulation of MIG production |
| Major function | Reduces the production of CXCL9, a chemokine that recruits CXCR3+ immune cells, thereby modulating immune responses and inflammation. |
| Related chemokine | CXCL9 (MIG), a member of the C-X-C chemokine family. |
| Key regulators | Type-I interferons, LINC00152/EZH2, autophagy pathways, and other transcriptional and post-transcriptional modulators. |
| Associated diseases | Cancer, autoimmune diseases, chronic inflammation, and angiogenesis-related disorders. |
| Research methods | CRISPR knockout, RNA-seq, ChIP-seq, ELISA, and chemotaxis assays. |
What Is GO:0035395?
GO:0035395, negative regulation of chemokine (C-X-C motif) ligand 9 production, is a biological process that encompasses any mechanism that decreases the synthesis, secretion, or overall availability of CXCL9. This includes transcriptional repression, post-transcriptional regulation, and inhibition of secretion. The term is synonymous with negative regulation of CXCL9 production and negative regulation of MIG production, reflecting the chemokine's alternative name.
Why Is negative regulation of chemokine (C-X-C motif) ligand 9 production Important in Cell Biology?
Negative regulation of CXCL9 production is crucial for maintaining immune balance and preventing pathological inflammation. CXCL9 is a potent chemoattractant for CXCR3+ T cells and NK cells, and its overproduction can lead to excessive immune cell infiltration, tissue damage, and autoimmune reactions. Conversely, insufficient CXCL9 production can impair antitumor immunity and pathogen clearance [5,8]. Therefore, understanding the mechanisms that negatively regulate CXCL9 is essential for developing therapies that fine-tune immune responses in cancer, autoimmunity, and infectious diseases.
• Prevents excessive immune cell recruitment and tissue damage in inflammatory diseases.
• Modulates antitumor immunity by controlling CD8+ T-cell infiltration in cancers such as gastric cancer.
• Regulates angiogenesis; inhibiting CXCL9 expression promotes angiogenesis in mesenchymal stem cells.
• Plays a role in wound healing through autophagy-mediated pathways in keratinocytes.
• Impacts autoimmune diseases like gouty arthritis by influencing Vδ2 T cell chemotaxis.
• Serves as a potential therapeutic target in breast cancer to modulate immune escape.
• Influences the efficacy of anti-PD-1/PD-L1 immunotherapy by affecting follicular helper T cells.
• Contributes to the immunomodulatory effects of lactic acid bacteria in mastitis.
• Provides a mechanism for cross-talk between autophagy and chemokine production.
• Helps maintain homeostasis in the central nervous system by suppressing microglial CXCL13 production.
What Happens During negative regulation of chemokine (C-X-C motif) ligand 9 production?
Transcriptional repression of CXCL9
In simple terms: Cells can turn down the CXCL9 gene by blocking the proteins that normally switch it on.
Transcriptional repression is a primary mechanism for negative regulation of CXCL9 production. Type-I interferons have been shown to suppress microglial production of the lymphoid chemokine CXCL13, and similar mechanisms may apply to CXCL9. In gastric cancer, the long non-coding RNA LINC00152 binds to EZH2, a component of the polycomb repressive complex 2, leading to epigenetic silencing of CXCL9 and reduced CD8+ T-cell infiltration. This demonstrates that negative regulation can occur through chromatin modifications and transcriptional interference.
Post-transcriptional and epigenetic regulation
In simple terms: Even after the CXCL9 gene is read, the message can be degraded or blocked before it makes protein.
Post-transcriptional mechanisms, including microRNA-mediated mRNA degradation and RNA-binding protein interference, can reduce CXCL9 production. Epigenetic modifications such as DNA methylation and histone deacetylation also contribute to long-term suppression of CXCL9 expression. These layers of regulation ensure that CXCL9 levels are tightly controlled in response to environmental cues.
Autophagy-mediated suppression
In simple terms: The cell's recycling system can influence how much CXCL9 is made.
Keratinocyte autophagy has been shown to enable the activation of keratinocytes and fibroblasts and facilitate wound healing. This process involves modulation of chemokine production, including CXCL9, suggesting that autophagy-related pathways can negatively regulate CXCL9 under certain conditions. The exact molecular links between autophagy and CXCL9 suppression are an active area of research.
Regulation by interferons and cytokines
In simple terms: Signals from the immune system can tell cells to stop making CXCL9.
Type-I interferons are known to suppress microglial production of CXCL13, and similar suppressive effects on CXCL9 have been observed in various cell types. In bovine mammary epithelial cells, lactic acid bacteria modulate immune responses, potentially affecting CXCL9 production. These findings highlight the role of cytokine networks in negatively regulating CXCL9.
Impact on immune cell chemotaxis
In simple terms: Less CXCL9 means fewer immune cells are called to the site.
By reducing CXCL9 production, negative regulation directly diminishes the chemotaxis of CXCR3+ cells, including activated T cells and NK cells. In acute gouty arthritis, Vδ2 T cell chemotaxis to the synovium is influenced by chemokine gradients, and negative regulation of CXCL9 could modulate this process. Similarly, in breast cancer, CXCL1 sustains cancer stem cell self-renewal and immune escape, and altering CXCL9 levels may impact these programs.
Key Genes Involved in GO:0035395 negative regulation of chemokine (C-X-C motif) ligand 9 production
The following genes and proteins are involved in the negative regulation of CXCL9 production, either as direct regulators or as components of upstream signaling pathways.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CXCL9 | Target chemokine; production is negatively regulated | Central to GO:0035395; its expression is modulated in cancer and inflammation [6,8]. |
| EZH2 | Epigenetic repressor; binds LINC00152 to silence CXCL9 | Involved in gastric cancer immune evasion. |
| LINC00152 | Long non-coding RNA; recruits EZH2 to CXCL9 promoter | Mediates CD8+ T-cell infiltration in gastric cancer. |
| IFNAR1 | Type-I interferon receptor; mediates suppressive signals | Suppresses microglial chemokine production. |
| IRF1 | Interferon regulatory factor; can activate or repress chemokines | Potential mediator of negative regulation. |
| STAT1 | Signal transducer; downstream of interferon signaling | May be involved in suppressing CXCL9. |
| ATG5 | Autophagy-related protein; enables keratinocyte autophagy | Links autophagy to chemokine modulation in wound healing. |
| ATG7 | Autophagy-related protein; essential for autophagosome formation | Potential role in negative regulation of CXCL9. |
| CXCR3 | Receptor for CXCL9; mediates chemotaxis | Readout for CXCL9 activity [3,8]. |
| CXCL10 | Related chemokine; co-regulated with CXCL9 | Shares regulatory pathways with CXCL9. |
| CXCL13 | Lymphoid chemokine; suppressed by type-I interferons | Model for negative regulation of chemokines. |
| NF-κB | Transcription factor; often activates chemokines | Its inhibition may lead to negative regulation. |
| JAK1 | Kinase; mediates interferon signaling | Upstream of STAT1 in suppressive pathways. |
| JAK2 | Kinase; mediates interferon signaling | Upstream of STAT1 in suppressive pathways. |
| SOCS1 | Suppressor of cytokine signaling; inhibits JAK-STAT | Potential negative regulator of CXCL9. |
| SOCS3 | Suppressor of cytokine signaling; inhibits JAK-STAT | Potential negative regulator of CXCL9. |
| miR-21 | MicroRNA; can target CXCL9 mRNA | Post-transcriptional regulation. |
| HDAC1 | Histone deacetylase; represses transcription | Epigenetic silencing of CXCL9. |
How Is negative regulation of chemokine (C-X-C motif) ligand 9 production Regulated?
The negative regulation of CXCL9 production is controlled by a complex network of signaling pathways. Type-I interferons, acting through the JAK-STAT pathway, can suppress CXCL9 production in microglia and other cells. The long non-coding RNA LINC00152 recruits EZH2 to the CXCL9 promoter, leading to histone methylation and transcriptional repression. Autophagy-related proteins such as ATG5 and ATG7 modulate chemokine production in keratinocytes, suggesting a link between cellular stress responses and CXCL9 regulation. Additionally, cytokines such as TGF-β and IL-10 can inhibit CXCL9 expression, although the exact mechanisms require further study. These regulatory mechanisms ensure that CXCL9 levels are appropriately tuned to the immune context.
negative regulation of chemokine (C-X-C motif) ligand 9 production and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CXCL9 | Gastric cancer immune evasion | LINC00152 knockout in gastric cancer cell lines |
| EZH2 | Cancer epigenetic silencing | EZH2 inhibitor treatment in cancer cells |
| CXCL9 | Angiogenesis | CXCL9 overexpression in MSCs-HUVECs co-culture |
| CXCL9 | Acute gouty arthritis | Vδ2 T cell chemotaxis assays |
| CXCL9 | Wound healing | Keratinocyte-specific autophagy knockout mice |
Cancer and immune evasion
Negative regulation of CXCL9 production is a key mechanism of immune evasion in cancer. In gastric cancer, LINC00152 binds EZH2 to repress CXCL9, reducing CD8+ T-cell infiltration and promoting tumor growth. Similarly, in breast cancer, CXCL1 sustains cancer stem cell self-renewal and immune escape, and modulation of CXCL9 may influence these processes. Follicular helper T cells can restore CD8+ T-cell-dependent antitumor immunity and anti-PD-L1/PD-1 efficacy, partly by regulating chemokine networks including CXCL9. These findings highlight the therapeutic potential of targeting negative regulators of CXCL9 to enhance immunotherapy.
Autoimmune and inflammatory diseases
In autoimmune conditions such as acute gouty arthritis, chemotaxis of Vδ2 T cells to the synovium is driven by chemokine gradients, and negative regulation of CXCL9 could modulate this pathogenic process. Type-I interferons suppress microglial production of lymphoid chemokines, which may protect against neuroinflammation. In mastitis, lactic acid bacteria modulate immune responses in bovine mammary epithelial cells, potentially affecting CXCL9 production. Dysregulation of CXCL9 negative regulation can thus contribute to chronic inflammation and tissue damage.
Wound healing and angiogenesis
Keratinocyte autophagy facilitates wound healing by modulating chemokine production, including CXCL9. Inhibiting CXCL9 expression in mesenchymal stem cells promotes angiogenesis in co-culture with HUVECs, suggesting that negative regulation of CXCL9 supports vascular remodeling. These roles implicate GO:0035395 in tissue repair and regenerative processes.
From negative regulation of chemokine (C-X-C motif) ligand 9 production-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does knockout of a candidate gene increase CXCL9 production? | CRISPR knockout cell lines (e.g., EZH2 KO) |
| Does a point mutation in a regulator affect CXCL9 repression? | CRISPR point mutation knock-in (e.g., STAT1 mutant) |
| Does overexpression of a repressor reduce CXCL9 levels? | CRISPR overexpression (e.g., LINC00152) |
| Does tagging a regulator alter its localization and function? | CRISPR knock-in of FLAG or GFP tag |
| Does autophagy modulate CXCL9 production? | ATG5 or ATG7 knockout keratinocytes |
| Does interferon signaling suppress CXCL9? | IFNAR1 knockout microglia |
How to Study the negative regulation of chemokine (C-X-C motif) ligand 9 production Process
| Method | What It Measures | Typical Application |
|---|---|---|
| RNA-seq | Global transcriptome changes | Identify genes co-regulated with CXCL9 |
| ChIP-seq | Histone modifications and transcription factor binding | Detect EZH2 binding at CXCL9 promoter |
| ELISA | CXCL9 protein concentration | Quantify suppression in cell supernatants |
| Western blot | Protein expression levels | Validate knockdown or knockout efficiency |
| Chemotaxis assay | Migration of CXCR3+ cells | Assess functional impact of CXCL9 regulation |
| CRISPR screen | Genes affecting CXCL9 production | Discover novel negative regulators |
| Flow cytometry | Immune cell infiltration | Measure CD8+ T-cell recruitment in tumors |
| Luciferase reporter | Promoter activity | Test transcriptional repression of CXCL9 |
Transcriptional profiling
RNA-seq and ChIP-seq can identify transcriptional changes and epigenetic marks at the CXCL9 locus upon negative regulation. For example, LINC00152-mediated recruitment of EZH2 to the CXCL9 promoter can be detected by ChIP-qPCR. These methods reveal the molecular mechanisms of repression.
Protein quantification
ELISA and Western blot are used to measure CXCL9 protein levels in cell culture supernatants and lysates. This is critical for confirming that negative regulation affects actual protein production, as shown in studies of microglial chemokine suppression.
Functional chemotaxis assays
Chemotaxis assays using CXCR3+ cells (e.g., activated T cells) measure the functional impact of altered CXCL9 production. Such assays have been used to study Vδ2 T cell migration in gouty arthritis and CD8+ T-cell infiltration in cancer.
CRISPR screening
Genome-wide CRISPR knockout or activation screens can identify novel regulators of CXCL9 production. These screens, combined with CXCL9 reporter systems, enable unbiased discovery of negative regulators.
How CRISPR Can Be Used to Study GO:0035395 negative regulation of chemokine (C-X-C motif) ligand 9 production
Knockout
CRISPR knockout of candidate negative regulators (e.g., EZH2, LINC00152) can be used to determine whether they are required for suppressing CXCL9 production. For instance, knocking out EZH2 in gastric cancer cells may increase CXCL9 levels and enhance CD8+ T-cell infiltration. Similarly, knocking out ATG5 in keratinocytes can reveal the role of autophagy in CXCL9 regulation.
Point Mutation
CRISPR point mutation knock-in can introduce specific amino acid changes in regulators to test their function. For example, mutating STAT1 phosphorylation sites can assess their role in interferon-mediated suppression of CXCL9. This approach provides mechanistic insights into signaling pathways.
Knock-in
CRISPR knock-in of tags (e.g., FLAG, GFP) or reporter genes (e.g., luciferase) into the CXCL9 locus allows real-time monitoring of CXCL9 production and regulation. Tagging endogenous CXCL9 enables tracking of its secretion and localization.
Overexpression
CRISPR activation (CRISPRa) or lentiviral overexpression can be used to overexpress negative regulators of CXCL9, such as LINC00152 or SOCS1, to confirm their suppressive effects. This is useful for validating gain-of-function phenotypes.
How EDITGENE Supports negative regulation of chemokine (C-X-C motif) ligand 9 production Research
Researchers studying negative regulation of chemokine (C-X-C motif) ligand 9 production-related genes often need to determine whether a candidate gene is causally involved in suppressing CXCL9. EDITGENE provides comprehensive CRISPR gene editing services to enable precise functional studies.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of chemokine (C-X-C motif) ligand 9 production research.
Frequently Asked Questions About negative regulation of chemokine (C-X-C motif) ligand 9 production
What is GO:0035395?
GO:0035395 is the Gene Ontology term for negative regulation of chemokine (C-X-C motif) ligand 9 production, describing any process that reduces the production of CXCL9.
What genes are involved in negative regulation of CXCL9 production?
Key genes include CXCL9 itself, EZH2, LINC00152, ATG5, ATG7, STAT1, and IFNAR1, among others [1,4,8].
How is CXCL9 production negatively regulated?
Mechanisms include transcriptional repression by EZH2/LINC00152, suppression by type-I interferons, and autophagy-mediated modulation [1,4,8].
Why is negative regulation of CXCL9 important in cancer?
It can reduce CD8+ T-cell infiltration and promote immune evasion, as seen in gastric cancer where LINC00152 represses CXCL9.
What diseases are associated with dysregulated CXCL9 production?
Cancer, autoimmune diseases like gouty arthritis, chronic inflammation, and angiogenesis-related disorders [3,6,8].
How can I study negative regulation of CXCL9 in the lab?
Use CRISPR knockout, RNA-seq, ChIP-seq, ELISA, and chemotaxis assays to measure CXCL9 levels and function [1,4,8].
Does autophagy affect CXCL9 production?
Yes, keratinocyte autophagy modulates chemokine production, including CXCL9, during wound healing.
What is the role of type-I interferons in CXCL9 regulation?
Type-I interferons suppress microglial production of lymphoid chemokines, including CXCL13, and likely CXCL9, through JAK-STAT signaling.
Can CRISPR be used to study negative regulation of CXCL9?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to dissect regulatory mechanisms.
What cell models are suitable for studying GO:0035395?
Gastric cancer cells, keratinocytes, microglia, mesenchymal stem cells, and bovine mammary epithelial cells are commonly used [1,4,6,7,8].
Conclusion
GO:0035395, negative regulation of chemokine (C-X-C motif) ligand 9 production, is a critical biological process that controls immune cell recruitment and inflammation. Dysregulation of this process contributes to cancer immune evasion, autoimmune diseases, and impaired wound healing. Understanding the genes and mechanisms involved, such as EZH2/LINC00152-mediated repression and autophagy, offers opportunities for therapeutic intervention. EDITGENE provides advanced CRISPR tools to accelerate research in this field.
References
- 1. Qiang L et al.. 2021. Keratinocyte autophagy enables the activation of keratinocytes and fibroblastsand facilitates wound healing.. Autophagy 17(9):2128-2143 PMID: 32866426
- 2. Ciummo SL et al.. 2021. The C-X-C Motif Chemokine Ligand 1 Sustains Breast Cancer Stem Cell Self-Renewal and Promotes Tumor Progression and Immune Escape Programs.. Front Cell Dev Biol 9:689286 PMID: 34195201
- 3. Di H et al.. 2024. Role of Chemotaxis of Vδ2 T Cells to the Synovium in the Pathogenesis of Acute Gouty Arthritis.. J Inflamm Res 17:721-736 PMID: 38344307
- 4. Esen N et al.. 2014. Type-I interferons suppress microglial production of the lymphoid chemokine, CXCL13.. Glia 62(9):1452-62 PMID: 24829092
- 5. Niogret J et al.. 2021. Follicular helper-T cells restore CD8(+)-dependent antitumor immunity and anti-PD-L1/PD-1 efficacy.. J Immunother Cancer 9(6) PMID: 34103351
- 6. Shen Q et al.. 2019. Inhibiting expression of Cxcl9 promotes angiogenesis in MSCs-HUVECs co-culture.. Arch Biochem Biophys 675:108108 PMID: 31550444
- 7. Fukuyama K et al.. 2020. Evaluation of the Immunomodulatory Ability of Lactic Acid Bacteria Isolated from Feedlot Cattle Against Mastitis Using a Bovine Mammary Epithelial Cells In Vitro Assay.. Pathogens 9(5) PMID: 32466097
- 8. Ou J et al.. 2021. LINC00152 mediates CD8(+) T-cell infiltration in gastric cancer through binding to EZH2 and regulating the CXCL9, 10/CXCR3 axis.. J Mol Histol 52(3):611-620 PMID: 33709190
