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Infection and Immunity, December 1999, p. 6611-6618, Vol. 67, No. 12
0019-9567/99/$04.00+0

Production of Tumor Necrosis Factor Alpha in Human T Lymphocytes by Staphylococcal Enterotoxin B Correlates with Toxin-Induced Proliferation and Is Regulated through Protein Kinase C

Zhengyin Yan,1,2,dagger David C. H. Yang,1 Roger Neill,2 and Marti Jett1,2,*

Chemistry Department, Georgetown University, Washington, D.C. 20056,1 and Department of Molecular Pathology, Walter Reed Army Institute of Research, Washington, D.C. 203072

Received 21 April 1999/Returned for modification 14 June 1999/Accepted 16 August 1999

The superantigen staphylococcal enterotoxin B (SEB) simultaneously binds both the major histocompatibility complex (MHC) class II receptor on monocytes and the T-cell receptor (TCR) on T lymphocytes, resulting in a range of cell responses including induction of tumor necrosis factor alpha (TNF-alpha ). In this study, we have used mixed cultures of human peripheral blood monocytes and lymphocytes to investigate biochemical events controlling SEB induction of TNF-alpha . TNF-alpha production induced by SEB in mixed cultures is more closely associated with T cells than with monocytes: (i) a TCR-binding-site mutant of SEB (N23F) is less active in TNF-alpha induction than an MHC class II receptor-binding-site mutant (F44R), and (ii) flow cytometric analysis indicated that SEB induced TNF-alpha production in T cells but not in monocytes. Pretreatment of cells with inhibitors of signal transduction pathways was employed to further define events in SEB-induced TNF-alpha production. Neither protein kinase A inhibitors nor two protein tyrosine kinase inhibitors altered SEB-induced TNF-alpha production. In contrast, SEB induced protein kinase C (PKC) translocation, and pretreatment of cultures with inhibitors of PKC blocked TNF-alpha induction. Alteration of levels of diacylglycerol (DAG), an activator of PKC, by treatment with inhibitors of phospholipase C or DAG kinase also altered SEB-induced TNF-alpha production. These data suggest that PKC activation plays a critical role in SEB-induced TNF-alpha production in human T cells.

* Corresponding author. Mailing address: Department of Molecular Pathology, Walter Reed Army Institute of Research, Building 503, 503 Robert Grant Rd., Silver Spring, MD 20910. Phone: (301) 319-9997. Fax: (301) 319-7699. E-mail: marti.jett{at}na.amedd.army.mil.

dagger Present address: R. W. Johnson Pharmaceutical Research Institute, Spring House, PA 19477.

Infection and Immunity, December 1999, p. 6611-6618, Vol. 67, No. 12
0019-9567/99/$04.00+0


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