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Expression of CD73/ecto‐5′‐nucleotidase on human gingival fibroblasts and contribution to the inhibition of interleukin‐1α‐induced granulocyte‐macrophage colony stimulating factor production
Author(s) -
Nemoto Eiji,
Kunii Ryotaro,
Tada Hiroyuki,
Tsubahara Taisuke,
Ishihata Hiroshi,
Shimauchi Hidetoshi
Publication year - 2004
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/j.1600-0765.2004.00698.x
Subject(s) - granulocyte macrophage colony stimulating factor , macrophage , microbiology and biotechnology , granulocyte , interleukin , granulocyte macrophage colony stimulating factor receptor , interleukin 1β , immunology , chemistry , biology , macrophage colony stimulating factor , cytokine , biochemistry , in vitro
Background and objectives: CD73/5′‐nucleotidase (5′‐NT) is an ectoenzyme that participates in immune/inflammatory reactions. We examined the possible expression of CD73/5′‐NT on human gingival fibroblasts (hGF), which are important to the immune/inflammatory system in periodontal tissue. Methods and results: We demonstrated that CD73/5′‐NT was expressed on hGF by flow cytometry. We found that pre‐treatment of hGF with 5′‐AMP induced marked inhibition of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) production from hGF upon stimulation with interleukin‐1α (IL‐1α) by enzyme‐linked immunosorbent assay (ELISA). A specific inhibitor of 5′‐NT, adenosine 5′‐[α,β‐methylene] diphosphate blocked the inhibition of GM‐CSF production, suggesting that adenosine converted from 5′‐AMP acts on the inhibitory effects. The GM‐CSF inhibition suggested that A3 receptor might be involved. The rank order of agonists was found to be ( N 6 ‐benzyl‐5′‐ N ‐ethylcarboxamidoadenosine) A3 receptor agonist ≥ (2‐chloroadenosine) non‐selective agonist > (CGS‐21680) A2 A receptor agonist > adenosine ≥ ( N 6 ‐cyclohexyladenosine) A1 agonist. Further support for the main role of A3 receptor was the binding A3 antagonist [9‐chloro‐2‐(2‐furanyl)‐5‐([phenylacetyl]amino)[1,2,4]‐triazolo[1,5‐ c ]quinazdine] reversed the effect of adenosine, but no significant reverse was observed by A1 (1,3‐dipropyl‐8‐cyclopentylxanthine), A2 [3,7‐dimethyl‐1‐(2‐propargyl)xanthine], A2 A [8‐(3‐chlorostyryl)caffeine], and A2 B (alloxazine) antagonists. The CD73/5′‐NT expression was increased upon stimulation with gamma‐interferon, but not other stimulants such as tumor necrosis factor‐alpha, IL‐4, lipopolysaccharide from Porphyromonas gingivalis and Escherichia coli , and fimbriae from P. gingivalis , and this increase was correlated with the enhanced GM‐CSF inhibition by 5′‐AMP but not adenosine. Conclusions: These findings suggested that CD73/5′‐NT on hGF exerts an anti‐inflammatory effects in periodontal disease by conversion from 5′‐AMP to adenosine.