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Divisions of
*
Pediatric Infectious Diseases and
Pediatric Intensive Care, Ahmanson Department of Pediatrics, Steven Spielberg Pediatric Research Center, Cedars-Sinai Medical Center, University of California School of Medicine, Los Angeles, CA 90048; and
Department of Immunology, Mayo Clinic, Rochester, MN 55905
In HIV-infected patients, concurrent infections with bacteria and
viruses are known to induce HIV replication as assessed by increases in
plasma HIV RNA levels. In the present study, we determined the cell
surface receptor and molecular mechanisms of enterobacterial
LPS-induced HIV transcription. Human dermal microvessel endothelial
cells (HMEC) were transfected with an HIV-long terminal repeat
(LTR)-luciferase construct and subsequently stimulated with purified
bacterial LPS. Our studies demonstrate that human Toll-like receptor 4
(TLR4) mediates LPS-induced NF-
B and HIV-LTR activation in HMEC
through IL-1 signaling molecules, namely myeloid differentiation
protein, IL-1R-associated kinase, TNFR-associated factor, and
NF-B-inducing kinase. Cotransfection of HMEC with HIV-LTR-luciferase
and TLR4 cDNA from LPS-hyporesponsive C3H/HeJ mice abrogates
LPS-induced HIV transcription as does the use of dominant-negative
mutants of the IL-1 signaling molecules. Transfection of HMEC with an
HIV-LTR-mutant that lacks the NF-B binding site or pretreatment of
cells with chemical inhibitors of the NF-B pathway also blocked
LPS-induced HIV-LTR transactivation. These data support the conclusion
that TLR4 mediates enterobacterial LPS-induced HIV transcription via
IL-1 signaling molecules and NF-B activation plays an important role
in HIV-LTR transactivation.
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