Increase of CD26/dipeptidyl peptidase IV expression on human gingival fibroblasts upon stimulation with cytokines and bacterial components.
CD26/dipeptidyl peptidase IV (DPPIV) is a cell surface ectoenzyme which participates in immune and inflammatory reactions. We found that CD26 was only partially expressed on human fibroblasts from periodontal tissues, whereas fibroblasts from lung and skin expressed CD26 constitutively as revealed by flow cytometry. We examined the possible upregulation of CD26 expression on human gingival fibroblasts in response to various stimulants. Interleukin-1alpha (IL-1alpha); tumor necrosis factor alpha; gamma interferon; lipopolysaccharide from Porphyromonas gingivalis, Prevotella intermedia, and Escherichia coli; and Prevotella glycoprotein augmented CD26 expression on gingival fibroblasts. Among the stimulants, IL-1alpha exhibited the most potent activity. Enzymatic activity of CD26 induced by IL-1alpha on fibroblasts was determined colorimetrically in terms of Gly-Pro hydrolysis of a synthetic chromogenic substrate, Gly-Pro p-nitroanilide. Among various inhibitors tested, diprotin A and phenylmethylsulfonyl fluoride inhibited the enzymatic activity, suggesting that the enzyme induced by IL-1alpha was DPPIV. The upregulation of CD26 mRNA expression upon stimulation with IL-1alpha was also revealed by a quantitative reverse transcription-PCR assay. In the kinetic experiment, 48 h and several days were required for maximum CD26 mRNA accumulation and CD26 molecule expression on the cell surface, respectively. The addition of cycloheximide at 2 h before IL-1alpha stimulation almost completely inhibited the accumulation of CD26 mRNA. These results suggested that induction of CD26 on human gingival fibroblasts is regulated at the transcriptional level and is also dependent on a de novo-synthesized protein factor(s).
Nemoto
E
E
Department of Endodontics and Periodontics, Tohoku University School of Dentistry, Sendai, 980-8575, Japan. eiji@mail.cc.tohoku.ac.jp
Sugawara
S
S
Takada
H
H
Shoji
S
S
Horiuch
H
H
D016207:Cytokines / Q000494:pharmacology*; D018819:Dipeptidyl Peptidase 4 / Q000096:biosynthesis*; D004790:Enzyme Induction; D005347:Fibroblasts / Q000201:enzymology / Q000276:immunology*; D005881:Gingiva / Q000166:cytology / Q000201:enzymology / Q000276:immunology*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008168:Lung / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D010513:Periodontal Ligament / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D018720:Prevotella / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000378:metabolism
D016207:Cytokines; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D018819:Dipeptidyl Peptidase 4