Applants Abstracts) This project will examine the effects of cocaine on HIV-1 infection of coronary artery endothelial cells (CAEC). Recent pivotal data suggest that HIV-1 infects CAEC and brain microvascular endothelial cells (BMVEC) in an abortive fashion leading to strong-stop DNA synthesis, cocaine increases this abortive infection with the JR-FL strain, and microvascular endothelial cells display modulation of CD4+, CXCR4 and CCR5. In CAEC and BMVEC, cocaine and HIV-1 induce IL-6 and cell adhesion molecules, and increases endothelial permeability. These effects may lead to microvascular endothelial leaks and to increased HIV-1 invasion and leukocyte transmigration into the heart interstitium. Preliminary studies: They have previously demonstrated that TNF-alpha and cocaine cause the microvascular endothelial barriers to allow HIV-1 invasion by a para- or transcellular route and have recently observed that HIV-1 infects CAEC abortively leading to reverse transcription of R/U5 without further steps of reverse transcription but with intracellular signaling, IL-6 induction and increase in cell permeability. They have noted that cocaine and inflammatory cytokines modulate the abortive infection by HIV-1JR-FL. They hypothesize that 1) the abortive infection and increased permeability of CAEC are mediated by cocaine's induction of IL-6 and modulation of CXCR4 and CCR5, and 2) the effects of the abortive HIV-1 infection, cocaine, and IL-6 result in an impairment of endothelial barrier function.
Specific Aims : They will determine 1) the effects of cocaine on the abortive infection of CAEC, 2) the effects of cocaine on passage and route of HIV-1 across CAEC, 3) cocaine's signaling, cocaine's signaling for IL-6, and the effects of cocaine, HIV-1 and IL-6 on CCR5 and CXCR4 expression, and 4) the effects of cocaine, HIV-1 and IL-6 on permeability, transendothelial electrical resistance, and procoagulant activity in CAEC.
