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Cord blood-derived human cultured mast cells produce transforming growth factor beta1
N Kanbe1, M Kurosawa, H Nagata
1Department of Dermatology, Gunma University School of Medicine, Maebashi, Japan.
Background:
Mast cells frequently accumulate at the site of fibrosis and their contribution has been suspected in the pathogenesis of fibrotic conditions. However, it still remains unknown whether human mast cells synthesize transforming growth factor beta1 (TGF-beta1).
Objective:
We have investigated whether cord blood-derived human cultured mast cells express messenger RNA (mRNA) for TGF-beta and produce bioactive TGF-beta1.
Methods:
Mast cells were obtained by culturing mononuclear cells from cord blood in the presence of stem cell factor and interleukin-6. Expression of mRNA for TGF-beta1 was examined by the method of reverse transcriptase-polymerase chain reaction (RT-PCR). Immunocytochemical staining for TGF-beta and growth-inhibitory assay using Mv1Lu cells were also performed.
Results:
The cultured human mast cells constitutively expressed mRNA for TGF-beta1. With calcium ionophore A23187, the intensity of the PCR-amplified band for TGF-beta1 was not increased. Immunocytochemical staining showed that the cultured mast cells were positive for both latency-associated peptide and activated forms of TGF-beta. Bioassay with Mv1Lu cells and R 4-2 mutant cells showed that mast-cell conditioned medium had a bioactivity of TGF-beta1.
Conclusion:
Cord blood-derived human cultured mast cells constitutively express mRNA for TGF-beta1 and produce functional TGF-beta1. Because TGF-beta1 has been shown to be highly fibrogenic, these results may highlight a novel role for human mast cells in tissue fibrosis.
Insights
Human mast cells, derived from cord blood, constitutively produce functional transforming growth factor beta1 (TGF-beta1). This finding suggests a new role for mast cells in the development of tissue fibrosis.
Area of Science:
- Immunology
- Cell Biology
- Fibrosis Research
Background:
- Mast cells are found at fibrotic sites, with suspected roles in pathogenesis.
- The synthesis of transforming growth factor beta1 (TGF-beta1) by human mast cells remains unconfirmed.
Purpose of the Study:
- To investigate if human mast cells from cord blood express mRNA for TGF-beta.
- To determine if these mast cells produce bioactive TGF-beta1.
Main Methods:
- Culturing mononuclear cells from cord blood to obtain mast cells.
- Utilizing reverse transcriptase-polymerase chain reaction (RT-PCR) for mRNA expression analysis.
- Performing immunocytochemical staining and bioassays to detect TGF-beta1 production.
Main Results:
- Cultured human mast cells constitutively express TGF-beta1 mRNA.
- Immunocytochemistry confirmed the presence of both latent and active TGF-beta forms.
- Mast cell conditioned medium demonstrated significant TGF-beta1 bioactivity.
Conclusions:
- Human mast cells derived from cord blood constitutively express and produce functional TGF-beta1.
- Given TGF-beta1's fibrogenic properties, these findings indicate a novel role for mast cells in tissue fibrosis.