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Clinical and Diagnostic Laboratory Immunology, March 2001, p. 325-332, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.325-332.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Butyric Acid-Induced T-Cell Apoptosis Is Mediated by Caspase-8 and -9 Activation in a Fas-Independent Manner

Tomoko Kurita-Ochiai,1,* Kuniyasu Ochiai,2 and Kazuo Fukushima1

Department of Microbiology, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba 271-8587,1 and Department of Oral Microbiology, Meikai University School of Dentistry, Sakado, Saitama 350-0283,2 Japan

Received 20 July 2000/Returned for modification 28 August 2000/Accepted 22 November 2000

Our previous study demonstrated that butyric acid, an extracellular metabolite of periodontopathic bacteria, induced apoptosis in murine thymocytes, splenic T cells, and human Jurkat cells. In this study, we examined whether CD95 ligand-receptor interaction is involved in butyric acid-induced T-cell apoptosis. Flow cytometry analysis indicated that expression of Fas in Jurkat and T cells from peripheral blood mononuclear cells was not affected by butyric acid treatment. Furthermore, the expression of Fas and FasL protein in Western blotting was not affected by butyric acid treatment. Coincubation with blocking anti-Fas antibodies prevented Fas-induced apoptosis but not butyric acid-induced apoptosis. Anti-FasL antibodies also did not prevent butyric acid-induced apoptosis at any dose examined. Although cytotoxic anti-Fas antibody affected butyric acid-induced apoptosis, a synergistic effect was not seen. Time-dependent activation of caspase-8 and -9 was recognized in butyric acid- as well as Fas-mediated apoptosis. IETD-CHO and LEHD-CHO, specific inhibitors of caspase-8 and -9, respectively, completely blocked Fas-mediated apoptosis and partially prevented butyric acid-induced apoptosis. These results suggest that the Fas-FasL interaction is not involved in butyric acid-induced apoptosis and that caspase-8 and -9-dependent apoptosis plays an important role in butyric acid-induced apoptosis, as well as Fas-induced apoptosis.

* Corresponding author. Mailing address: Department of Microbiology, Nihon University School of Dentistry at Matsudo, Matsudo-shi, Chiba 271-8587, Japan. Phone and fax: 47-360-9343. E-mail: tkurita{at}mascat.nihon-u.ac.jp.

Clinical and Diagnostic Laboratory Immunology, March 2001, p. 325-332, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.325-332.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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