Involvement of CD73 (ecto-5'-nucleotidase) in adenosine generation by human gingival fibroblasts.

Adenosine has various biological effects on human gingival fibroblasts (HGF) and epithelial cells closely associated with inflammation, such as cytokine production and cell adhesion. However, the mechanism of adenosine formation in periodontal tissues is not yet defined. In this study, we examined the involvement of CD73 (ecto-5'-nucleotidase) in adenosine generation by HGF. CD73 was detected on in vitro-maintained HGF by immunocytochemistry and flow cytometric analysis. Adenosine production was observed following the addition of 5'-AMP, the substrate of CD73-associated ecto-5'-nucleotidase. Moreover, the addition of 5'-AMP to cultured HGF resulted in the elevation of cyclic adenosine monophosphate (cAMP). The 5'-AMP-induced increase in intracellular cAMP level was inhibited markedly by xanthine amine congener, an adenosine receptor antagonist, and partially by alpha,beta-methylene adenosine 5'-diphosphate, an ecto-5'-nucleotidase inhibitor. These results suggest that CD73 on HGF is a critical enzyme responsible for the generation of adenosine, an immunomodulator that activates adenosine receptors.