Abstract
Current antiretroviral drugs (ART) for HIV-1 need to be taken every day. As a result, treatment strategies are often unsuccessful due to lack of resources or drug fatigue. However, nanoparticles may serve as an improved drug carrier system for HIV-1 treatment. In these studies antiretroviral nanoparticles (ART-NP) have been designed to carry three different antiretroviral drugs, ritonavir, lopinavir. and efavirenz. The use of three drugs is important to reduce potential drug resistance by HIV-1. Past experiments have shown that ART-NP can continually release the three drugs in PBMC cells for up to 28 days, whereas free ART drugs only remained in the cells for 48 hours. We have assessed viability of immune cells which are the targets of HIV-1 infection following treatment with ART-NPs. Our data show no significant changes in viability or immune function as assessed by CellTitre Glow and EI.ISA analysis. Western blot analysis is currently being used to determine whether treatment with ART-NP can reduce HIV-1 load following infection in immune cells. We hypothesize that the HIV-1 proteins will be reduced in cells treated with ART-NP. The U937 human monocytic cell line, and the H9 human T cell line were infected with HIV-1 NL4-3. Following infection, cells were treated with ART-NP, free ART drug, blank nanoparticles, or left untreated. Subcellular fractionation of cellular lysates was used to separate cytoplasmic, membrane, nuclear, cytoskeletal, and chromatin-bound nuclear cellular materials. Western blot analysis of p17, p24, and gp120 viral proteins will be used to determine whether ART-NP treatment reduces viral replication in infected cells. HPLC will be done on the extracts to determine in which part of the cell ART drug is most concentrated. Future experiments will be directed at determining whether ART-NPs act prophylactically by blocking HIV-1 infection of immune cells. The evidence obtained from this project will be useful to help determine whether ART-NP will be useful as a therapeutic agent for HIV-1.