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Infection and Immunity, March 2002, p. 1049-1055, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1049-1055.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Essential Role of Interleukin-12 (IL-12) and IL-18 for Gamma Interferon Production Induced by Listeriolysin O in Mouse Spleen Cells

Takamasa Nomura, Ikuo Kawamura, Kohsuke Tsuchiya, Chikara Kohda, Hisashi Baba, Yutaka Ito, Terumi Kimoto, Isao Watanabe, and Masao Mitsuyama*

Department of Microbiology, Kyoto University Graduate School of Medicine, Sakyo-ku, Kyoto 606-8501, Japan

Received 9 July 2001/ Returned for modification 22 August 2001/ Accepted 14 November 2001

The mechanism of gamma interferon (IFN-{gamma}) production induced by listeriolysin O (LLO), a cytolytic virulence factor of Listeria monocytogenes, was analyzed with special reference to the involvement of macrophage-derived cytokines in spleen cells of mice. LLO purified from the culture supernatant of L. monocytogenes was capable of inducing a high level of IFN-{gamma} when its cytolytic activity was blocked by cholesterol treatment. The IFN-{gamma}-inducing ability of LLO was not dependent on possibly contaminating lipopolysaccharide. Depletion of CD11b+ cells resulted in a profound decrease in IFN-{gamma} production in response to LLO stimulation. Negative selection also suggested the contribution of DX5+ cells in IFN-{gamma} production. Reverse transcription-PCR revealed that expression of interleukin-12 (IL-12) p35 and p40 was induced by LLO but that the IL-18 mRNA level in the CD11b+ fraction of spleen cells was unchanged. There was no change in the expression of the IFN-{gamma}-inducing cytokine genes in the CD11b- fraction. Neutralization of IL-12 and IL-18 in culture abolished the IFN-{gamma} production almost completely. Spleen cells from IL-12- or IL-18-deficient mice never produced IFN-{gamma} after stimulation with LLO. These results clearly indicated that LLO, a well-known virulence factor of L. monocytogenes, is capable of inducing IFN-{gamma} from NK cells through induction of IL-12 and IL-18 from macrophages. LLO appeared to play essential roles, not only as a bacterial virulence factor but also as a bacterial modulin in the immune response of the host.

* Corresponding author. Mailing address: Department of Microbiology, Kyoto University Graduate School of Medicine, Yoshida konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan. Phone: 81-75-753-4441. Fax: 81-75-753-4446. E-mail: mituyama{at}mb.med.kyoto-u.ac.jp.

Infection and Immunity, March 2002, p. 1049-1055, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1049-1055.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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