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The Journal of Immunology, 2007, 179: 6910-6918.
Copyright © 2007 by The American Association of Immunologists, Inc.
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CCAAT/Enhancer-Binding Protein β and {delta} Binding to CIITA Promoters Is Associated with the Inhibition of CIITA Expression in Response to Mycobacterium tuberculosis 19-kDa Lipoprotein1

Meghan E. Pennini*, Yi Liu*, Jianqi Yang{dagger}, Colleen M. Croniger{ddagger}, W. Henry Boom*,§ and Clifford V. Harding2,*

* Department of Pathology, {dagger} Department of Biochemistry, {ddagger} Department of Nutrition, and § Department of Division of Infectious Diseases and Tuberculosis Research Unit, Case Western Reserve University, Cleveland, OH 44106

TLR2 signaling by Mycobacterium tuberculosis 19-kDa lipoprotein (LpqH) inhibits IFN-{gamma}-induced expression of CIITA by macrophages. Microarray analysis, quantitative RT-PCR, and Western blots showed that LpqH induced C/EBPβ and C/EBP{delta} in kinetic correlation with inhibition of CIITA expression. Of the C/EBPβ isoforms, liver inhibitory protein (LIP) was notably induced and liver-activating protein was increased by LpqH. Putative C/EBP binding sites were identified in CIITA promoters I and IV (pI and pIV). LpqH induced binding of C/EBPβ (LIP and liver-activating protein) to biotinylated oligodeoxynucleotide containing the pI or pIV binding sites, and chromatin immunoprecipitation showed that LpqH induced binding of C/EBPβ and C/EBP{delta} to endogenous CIITA pI and pIV. Constitutive expression of C/EBPβ LIP inhibited IFN-{gamma}-induced CIITA expression in transfected cells. In summary, LpqH induced expression of C/EBPβ and C/EBP{delta}, and their binding to CIITA pI and pIV, in correlation with inhibition of IFN-{gamma}-induced expression of CIITA in macrophages, suggesting a role for C/EBP as a novel regulator of CIITA expression.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 This work was supported by National Institutes of Health Grants AI035726 and AI034343 (to C.V.H.) and AI027243 and HL055967 (to W.H.B.).

2 Address correspondence and reprint requests to Dr. Clifford V. Harding, Department of Pathology, Wolstein 6534, Case Western Reserve University, 2103 Cornell Road, Cleveland, OH 44106-7288. E-mail address: cvh3{at}cwru.edu

3 Abbreviations used in this paper: Mtb, Mycobacterium tuberculosis; CD-RAP, cartilage-derived retinoic acid-sensitive protein; ChIP, chromatin immunoprecipitation; IRF, IFN regulatory factor; LAP, liver-activating protein; LIP, liver inhibitory protein; LpqH, Mtb 19-kDa lipoprotein; ODN, oligodeoxynucleotide; pI, CIITA promoter I; pIV, CIITA promoter IV; TX114, Triton X-114.






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