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Originally published In Press as doi:10.1074/jbc.M002482200 on May 22, 2000

J. Biol. Chem., Vol. 275, Issue 31, 23674-23684, August 4, 2000
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Involvement of STAT-1 and Ets Family Members in Interferon-gamma Induction of CD40 Transcription in Microglia/Macrophages*

Vince T. NguyenDagger and Etty N. Benveniste§

From the Department of Cell Biology, The Univeristy of Alabama at Birmingham, Birmingham, Alabama 35294-0005

Cluster of differentiation (CD)-40 is a cell surface receptor belonging to the tumor necrosis factor receptor family that plays a critical role in the regulation of immune responses. We have previously shown that the cytokine interferon (IFN)-gamma induces CD40 expression in microglia. Herein, we have elucidated the molecular mechanisms underlying IFN-gamma induction of CD40 gene expression in microglia/macrophages. IFN-gamma up-regulates CD40 expression at the transcriptional level, and this regulation involves the STAT-1alpha transcription factor. Microglia from STAT-1alpha -deficient mice were refractive to IFN-gamma induction of CD40 expression, illustrating the importance of STAT-1alpha in this response. Functional analysis of the CD40 promoter indicates that two gamma activated sequence elements as well as two Ets elements are involved in IFN-gamma induction of CD40 promoter activity. STAT-1alpha binds to the gamma activated sequence elements, whereas PU.1 and/or Spi-B bind to the Ets elements. The expression of PU.1 and Spi-B, in conjuction with STAT-1alpha activation, correlates with IFN-gamma inducibility of CD40 expression. Collectively, our data demonstrate the involvement of STAT-1alpha , PU.1, and Spi-B in IFN-gamma induction of CD40 gene expression in cells of the macrophage lineage.

* This work was supported by National Multiple Sclerosis Society Grant RG-2205-B-9 (to E. N. B.), National Institutes of Health Grant NS-36765 (to E. N. B.), and Grant AM-20614 from the University of Alabama at Birmingham Flow Cytometry Core Facility.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Supported by a National Institutes of Health Predoctoral Fellowship T32AI-07493.

§ To whom correspondence should be addressed: Dept. of Cell Biology, MCLM 350, The University of Alabama at Birmingham, 1918 University Blvd., Birmingham, AL 35294-0005. Tel.: 205-934-7667; Fax: 205-975-6748; E-mail: tika@uab.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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