Inhibition of HIV-1 in cell culture by oligonucleotide-loaded nanoparticles.
OBJECTIVE
To investigate the potential use of polymeric nanoparticles for the delivery of antisense oligonucleotides in HIV-1-infected cell cultures.


METHODS
Phosphorothioate oligonucleotides were encapsulated into poly (D,L-lactic acid) nanoparticles. Two models of infected cells were used to test the ability of nanoparticles to deliver them. HeLa P4-2 CD4+ cells, stably transfected with the beta-galactosidase reporter gene, were first used to evaluate the activity of the oligonucleotides on a single-round infection cycle. The acutely infected lymphoid CEM cells were then used to evaluate the inhibition of the viral production of HIV-1 by the oligonucleotides.


RESULTS
The addition to infected CEM cells of nanoparticles containing gag antisense oligonucleotides in the nanomolar range led to strong inhibition of the viral production in a concentration-dependent manner. Similar results were previously observed in HeLa P4-2 CD4+ cells. Nanoparticle-entrapped random-order gag oligonucleotides had similar effects on reverse transcription. However, the reverse transcriptase activity of infected cells treated with nanomolar concentrations of free antisense and random oligonucleotides was not affected.


CONCLUSIONS
These results suggest that poly (D,L-lactic acid) nanoparticles may have great potential as an efficient delivery system for oligonucleotides in HIV natural target cells, i.e., lymphocytic cells.
Berton
M
M
School of Pharmacy, University of Geneva, Switzerland. myriam.berton@pharm.unige.ch
Turelli
P
P
Trono
D
D
Stein
C A
CA
Allémann
E
E
Gurny
R
R
D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004339:Drug Compounding; D005079:Excipients; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D007202:Indicators and Reagents; D019344:Lactic Acid; D008214:Lymphocytes / Q000187:drug effects / Q000821:virology; D008863:Microspheres; D009841:Oligonucleotides / Q000008:administration & dosage* / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology; D011091:Polyesters; D011108:Polymers; D018894:Reverse Transcriptase Inhibitors / Q000008:administration & dosage* / Q000494:pharmacology*; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects
D005079:Excipients; D007202:Indicators and Reagents; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D011091:Polyesters; D011108:Polymers; D018894:Reverse Transcriptase Inhibitors; D019344:Lactic Acid; C033616:poly(lactide); D054303:HIV Reverse Transcriptase