Cellular uptake of ODNs in HIV-1 human-infected cells: a role for viral particles in DNA delivery?
We have previously described how a 16 nucleotides ODN (termed 93del) is capable of inhibiting the activity of recombinant integrase in a cell-free system as well as HIV-1 replication in human-infected cells with IC(50) in the low nanomolar range. Intracellular HIV-1 replication was inhibited when the ODN was added at the onset of infection. These results raise several questions. Is a naked ODN able to enter the cell? Does the virus play a role in ODN entry? The uptake of several ODNs (93del, 60del(sc), TBA, T30923) was evaluated and then tracked by labeling the ODN with a fluorescent dye and assessing its intracellular localization by confocal microscopy. A significant level of cellular uptake of free ODN was observed in several cell lines: HeLa epithelial cells, Huh7 hepatic cells, and H9 lymphocytes, and was detected for all ODNs tested except for TBA. Striking differences were observed when naked ODNs were added to cell in the presence or absence of the virus. When HIV-1 virions were present a sharp increase in cellular fluorescence was observed. These results strongly suggest a role for HIV-1 virions in the uptake of certain ODNs.
Métifiot
Mathieu
M
UMR 5097 CNRS, Université Victor Segalen Bordeaux 2, 33076 Bordeaux cedex, France.
Faure
Aurélie
A
Guyonnet-Duperat
Véronique
V
Bellecave
Pantxika
P
Litvak
Simon
S
Ventura
Michel
M
Andréola
Marie-Line
ML
D002460:Cell Line; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D019427:HIV Integrase / Q000378:metabolism; D019428:HIV Integrase Inhibitors / Q000494:pharmacology; D015497:HIV-1 / Q000378:metabolism / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D009841:Oligonucleotides / Q000378:metabolism* / Q000494:pharmacology; D014771:Virion / Q000502:physiology
D005456:Fluorescent Dyes; D019428:HIV Integrase Inhibitors; D009841:Oligonucleotides; D019427:HIV Integrase