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(C/EBP) Activates CCR5 Promoter: Increased C/EBP and CCR5 in T Lymphocytes from HIV-1-Infected Individuals
Human Retrovirus Section, Basic Research Laboratory, National Cancer Institute- Frederick, Frederick, MD 21702
C/EBP
is a member of a family of leucine zipper transcription
factors that are involved in regulating the expression of several
cytokines, including IL-1, IL-6, IL-8, TNF, and macrophage-inflammatory
protein-1
. We identified multiple C/EBP binding sites within the
gene for CCR5, suggesting that C/EBP may be involved in its
regulation. Transient transfection experiments in both myeloid and
lymphoid cells showed an increase in CCR5 promoter-driven green
fluorescent protein production in the presence of C/EBP. Deletion
analysis identified two C/EBP-responsive regions in the
CCR5 gene, one in the promoter region and one at
the 3' part of the intron. We provide evidence that, in myeloid cells
(U937), C/EBP independently activates CCR5 expression through sites
located either in the promoter region or in the intron of the
CCR5 gene. In contrast, in lymphoid cells (Jurkat) the
presence of the intronic cis-regulatory regions is
required for C/EBP-mediated activation. In agreement with the
functional data, EMSA demonstrated that in both myeloid and lymphoid
cells C/EBP binds specifically to sites present in the intron,
whereas interaction with the sites located in the promoter was cell
type specific and was detected only in myeloid cells. Analysis of
C/EBP in primary PBMCs obtained from HIV-1-infected individuals
revealed a significant increase in C/EBP expression. The enhanced
C/EBP activity correlated with a higher frequency of circulating
CCR5+ lymphocytes in AIDS patients and with a decline in
CD4 lymphocyte numbers. Taken together, these results suggest that
C/EBP is an important regulator of CCR5 expression and may play a
relevant role in the pathogenesis of HIV
disease.
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