AEGiS-15IAC: Host cell gene expression during HIV-1 latency and reactivation.

15th International AIDS Conference

Bangkok, Thailand - July 11-16, 2004

Host cell gene expression during HIV-1 latency and reactivation.

Int Conf AIDS 2004 Jul 11-16; 15:(abstract no. ThOrA1397)

Zeichner S, Krishnan V
HIV and AIDS Malignancy Branch, NCI, NIH, Bethesda, United States

BACKGROUND: To better understand cellular machinery involved in HIV lytic replication and latency, we studied cellular gene expression before and during reactivation and completion of the lytic viral cycle in HIV-1 chronically infected cells.

METHODS: Cellular gene expression profiles were examined with cDNA ~10K element human gene arrays and fluorescently labeled cDNA made from ACH-2, J1.1, and U1 chronically infected cells and uninfected parental lines. RNA quantitation was also done with realtime RT-PCR. Reactivation following selected treatments was assessed via p24 ELISA.

RESULTS: After lytic cycle induction, we observed ordered, time-dependent changes in cell gene expression patterns. About 1740 genes, in 220 known pathways, were differentially expressed (p< 0.001), indicating that completion of HIV lytic replication is associated with distinct, temporally-ordered host cell gene expression changes. Maximum changes were observed in early and intermediate phases of the cycle. We also found several genes with altered expression before induction. We studied expression profiles of three chronically infected cell lines to determine whether all the uninduced cells showed similar changes. About 40 genes were found to be significantly altered in all three cell lines, including genes previously linked to HIV replication (e.g. cdc42 and Lyn) and genes previously not associated with HIV. Several gene classes showed increased expression before lytic replication, notably genes encoding proteasomes, histone deacetylases, and transcription factors. To determine the effects of targeting products of genes differentially expressed in latently infected cells, we treated latently infected cells with a proteasome inhibitor, clastolactacystin-beta-lactone (CLBL), and an Egr1 activator, resveratrol, and found that the agents activated lytic replication.

CONCLUSIONS: Our results may offer new insights into the interaction of latently infected host cells and HIV, and suggest therapeutic approaches for inhibiting HIV infection and manipulating latently infected cells.

Keywords: AEGIS, HIV-1, Proteasome Endopeptidase Complex, Virus Replication, Gene Expression, HIV Infections, Acquired Immunodeficiency Syndrome, HIV Seropositivity, Gene Expression Regulation, Viral, Reverse Transcriptase Polymerase Chain Reaction, HIV-1 Reverse Transcriptase, Cell Line, Oligonucleotide Array Sequence Analysis, Humans, virology, genetics

040711
ThOrA1397