The role of IL-1β in ectonucleotidases and adenosine receptors expression by human gingival fibroblasts – preliminary results
Introduction/Context/Diagnosis
Adenosine tri-phosphate (ATP) is strongly involved in inflammation not only by acting on cells of the immune system but also by exerting its activity on cells of the microenvironment. During inflammation, stressed cells can release ATP to the extracellular medium and this ATP can be hydrolyzed to adenosine by the action of ectonucleotidases, such as CD39 and CD73. It was previously reported that ATP inhibits IL-1β-induced matrix metalloproteinases through the action of CD39 in primary human gingival fibroblasts (HGF). Also, it has been documented that CD73 expression in HGF is involved in adenosine production by HGF. However, little information is available on the influence of IL-1 in the expression of CD39, CD73, as well as adenosine receptors (ADORA) in HGF. Here, we hypothesize that IL-1 signaling is involved in the CD39, CD73, and adenosine receptor expression by HGF.
Methods/Treatment Plan
Primary human gingival fibroblasts were isolated from gingival explants of healthy patients after obtaining informed consent. Gingival tissue was cut into small pieces and then cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% heatinactivated fetal bovine serum and antibiotics. Cells were used in passage 5 as sub confluent monolayers for experiments. After 2h, 4h or 6h of IL-1β stimulation (1ng/mL) total cellular RNA was extracted and reverse transcribed for quantitative polymerase chain reaction (qPCR) experiments. We evaluated mRNA expression of CD39, CD73, adenosine receptors (ADORA) A2a and A2b.
Results/Outcome
Even though neither CD39 nor CD73 were modulated after IL-1β stimulation, adenosine receptors showed increased mRNA levels. IL-1β induced peak mRNA expression for both A2a and A2b after 4h.
Significance/Conclusions
This project has just started to unveil the role of IL-1β stimulation in the expression of important ectonucleotidases involved in the ATP breakdown in the inflammatory environment. It is expected that these results will contribute to a better understanding of how purinergic signaling can influence the response of numerous cells of the connective tissue.
Location
University of the Pacific, Dugoni Dental School, San Francisco, CA
Format
Poster
Poster Session
1st and 2nd Year Student Research Presentations
The role of IL-1β in ectonucleotidases and adenosine receptors expression by human gingival fibroblasts – preliminary results
University of the Pacific, Dugoni Dental School, San Francisco, CA
Adenosine tri-phosphate (ATP) is strongly involved in inflammation not only by acting on cells of the immune system but also by exerting its activity on cells of the microenvironment. During inflammation, stressed cells can release ATP to the extracellular medium and this ATP can be hydrolyzed to adenosine by the action of ectonucleotidases, such as CD39 and CD73. It was previously reported that ATP inhibits IL-1β-induced matrix metalloproteinases through the action of CD39 in primary human gingival fibroblasts (HGF). Also, it has been documented that CD73 expression in HGF is involved in adenosine production by HGF. However, little information is available on the influence of IL-1 in the expression of CD39, CD73, as well as adenosine receptors (ADORA) in HGF. Here, we hypothesize that IL-1 signaling is involved in the CD39, CD73, and adenosine receptor expression by HGF.
Comments/Acknowledgements
This work was supported by Research Grants to ACM Award 01-Activity 333 from the Arthur A. Dugoni School of Dentistry.