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STAT2-IRF1-ISG15-Associated Temporal Immune Reprogramming in Macrophages during Aspergillus fumigatus Infection
Quan Zhou1,2, Rendong Li3, Dandan Song1,2
1Department of Dermatology, The First Affiliated Hospital of Nanchang University, Nanchang, China.
Abstract:
Aspergillus fumigatus poses a critical threat to immunocompromised hosts through invasive aspergillosis, with mortality rates reaching 50-90%. Although macrophages initiate early antifungal defenses via pattern recognition receptors, the temporal dynamics of immune reprogramming remain incompletely characterized. This study integrated time-series transcriptomics (GSE202286) of human monocyte-derived macrophages exposed to A. fumigatus conidia (0-8 h) with experimental validation to delineate immune-phase transitions and identify candidate regulatory modules. Bioinformatics analysis revealed a progressive increase in differential gene expression, peaking at 8 h (2,636 upregulated and 1,940 downregulated genes). Gene set enrichment analysis (GSEA) showed early enrichment of TNF-α/NF-κB signaling (NES = 2.37, 2 h), followed by a prominent interferon-γ response at 8 h (NES = 2.37) that coincided with ongoing inflammatory pathway activity. Temporal clustering identified 448 dynamically regulated genes associated with oxidative stress response (GO:0006979) and exploratory C-type lectin receptor-related pathway patterns (KEGG hsa04625). Protein interaction and transcription factor analyses predicted STAT2 as a candidate upstream regulator associated with IRF1 and ISG15. qRT-PCR validation in THP-1-derived macrophages showed sequential mRNA induction, with STAT2 peaking at 2 h, IRF1 at 4 h, and ISG15 remaining elevated through 8 h. A representative Western blot further supported this temporal pattern, showing early STAT2 phosphorylation, subsequent IRF1 protein induction, and later accumulation of free ISG15. Together, these findings suggest a STAT2-IRF1-ISG15-associated transcriptional program that may contribute to the transition from an inflammatory to an interferon-augmented macrophage response during A. fumigatus challenge.
Insights
This study reveals how macrophages transition their immune response to Aspergillus fumigatus, identifying a STAT2-IRF1-ISG15 pathway crucial for antifungal defense in immunocompromised individuals.
Area of Science:
- Immunology
- Molecular Biology
- Bioinformatics
Background:
- Invasive aspergillosis by Aspergillus fumigatus is a severe threat to immunocompromised individuals, with high mortality.
- Macrophages initiate early defense against A. fumigatus, but the dynamics of their immune reprogramming are not fully understood.
Purpose of the Study:
- To characterize the temporal immune reprogramming of macrophages upon exposure to A. fumigatus.
- To identify key regulatory modules and signaling pathways involved in macrophage antifungal defense.
Main Methods:
- Time-series transcriptomics of human monocyte-derived macrophages exposed to A. fumigatus conidia (0-8 hours).
- Bioinformatics analyses including gene set enrichment analysis (GSEA) and temporal clustering.
- Experimental validation using qRT-PCR and Western blotting in THP-1 derived macrophages.
Main Results:
- Gene expression progressively increased, peaking at 8 hours with thousands of upregulated and downregulated genes.
- Early TNF-α/NF-κB signaling was followed by a prominent interferon-γ response at 8 hours.
- A STAT2-IRF1-ISG15 transcriptional program was identified, with sequential protein induction and phosphorylation.
Conclusions:
- The study elucidates a STAT2-IRF1-ISG15 associated transcriptional program in macrophages during A. fumigatus challenge.
- This program may mediate the shift from an initial inflammatory response to an interferon-augmented defense.
- Findings provide insights into macrophage immune reprogramming critical for combating invasive aspergillosis.
