Involvement of STAT-1 and Ets family members in interferon-gamma inductionof CD40 transcription in microglia/macrophages

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 ha ve elucidated the molecular mechanisms underlying IFN-gamma induction of CD 40 gene expression in microglia/macrophages. IFN-gamma up-regulates CD40 ex pression at the transcriptional level, and this regulation involves the STA T-1 alpha transcription factor. Microglia from STAT-1 alpha-deficient mice were refractive to IFN-gamma induction of CD40 expression, illustrating the importance of STAT-1 alpha in this response. Functional analysis of the CD 40 promoter indicates that two gamma activated sequence elements as well as two Ets elements are involved in IFN-gamma induction of CD40 promoter acti vity. STAT-1 alpha 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-1 alpha activation, correlates with IFN-gamma in ducibility of CD40 expression. Collectively, our data demonstrate the invol vement of STAT-1 alpha, PU.1, and Spi-B in IFN-gamma induction of CD40 gene expression in cells of the macrophage lineage.