Related Experiment Videos
Modulation of cytokine expression in mouse dendritic cell clones
F Granucci1, G Girolomoni, M B Lutz
1Department of Pharmacology, CNR Center of Cytopharmacology, University of Milan, Italy.
Insights
Dendritic cells (DCs) exhibit limited cytokine gene expression compared to macrophages. Granulocyte/macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) differentially regulate cytokine production in distinct DC clones.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are crucial for initiating immune responses.
- Cytokine production by DCs remains incompletely understood.
- Immortalized DC clones offer a model to study DC function.
Purpose of the Study:
- To characterize the cytokine gene expression profile of two distinct immortalized mouse dendritic cell clones.
- To investigate the effects of granulocyte/macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) on DC cytokine production.
- To compare the cytokine expression patterns of DCs with those of macrophages.
Main Methods:
- Generation and immortalization of two mouse spleen-derived dendritic cell clones (CB1 and D2SC/1).
- Treatment of DC clones with GM-CSF, LPS, and interferon-gamma (IFN-γ).
- Analysis of cytokine gene expression (mRNA transcription) and protein synthesis.
Main Results:
- Transforming growth factor-beta 1 (TGF-β1) was constitutively transcribed, with detectable production only in D2SC/1 cells upon GM-CSF treatment.
- GM-CSF induced IL-1β transcription and synthesis in CB1 cells, enhancing their antigen presentation capacity.
- LPS upregulated gene expression and induced tumor necrosis factor-alpha (TNF-α) release in both clones, and additionally induced IL-1α and IL-1β in D2SC/1 cells.
- IFN-γ augmented antigen presentation but did not induce cytokine gene expression; IL-4, IL-10, and IL-12 mRNA were not induced by any stimulus.
Conclusions:
- Dendritic cells possess a restricted cytokine gene expression profile compared to macrophages.
- The two DC clones exhibit heterogeneity in their functional properties and responses to stimuli.
- Cytokine production by DCs is differentially regulated by specific stimuli like GM-CSF and LPS.
Abstract:
Dendritic cells (DC) play an essential role in the induction of primary immune responses; however, very little information is available on cytokine production by DC. Here we determined the cytokine gene expression profile of two immortalized DC clones, CB1 and D2SC/1, both generated from mouse spleen but differing in their activation requirements. Among the cytokines tested, only transforming growth factor-beta 1 was transcribed constitutively, but its production was detected only in D2SC/1 cells after treatment with granulocyte/macrophage colony-stimulating factor (GM-CSF). GM-CSF also promoted transcription and synthesis of interleukin (IL)-1 beta in CB1 cells that need pretreatment with GM-CSF to present major histocompatibility complex class II-restricted antigens efficiently in vitro. Lipopolysaccharide (LPS) up-regulated gene expression and induced release of tumor necrosis factor-alpha in both DC clones. In addition, LPS induced transcription of IL-1 alpha and both gene expression and synthesis of IL-1 beta in D2SC/1 cells. Interferon-gamma was ineffective in inducing cytokine gene expression, although it augmented the antigen-presentation capacity of DC, IL-4, IL-10 and IL-12 mRNA were not induced by any of the tested stimuli. The results suggest that DC have a limited cytokine gene expression pattern compared to macrophages and are heterogenous in some functional properties.