title: Pre-incubation of cell-free HIV-1 group M isolates with non-nucleoside reverse transcriptase inhibitors blocks subsequent viral replication in co-cultures of dendritic cells and T cells.
abstract: In order to study the inhibitory effect of various reverse transcriptase inhibitors (RTIs) on cell-free HIV, we adapted a recently described in vitro system, based on co-cultures of dendritic cells and resting CD4 T cells, modelling early target cells during sexual transmission. The compounds tested included the second-generation non-nucleoside RTI (NNRTI) TMC-120 (R147681, dapivirine) and TMC-125 (R165335, travertine), as well as the reference nucleoside RTI AZT (zidovudine), the nucleotide RTI PMPA (tenofovir) and the NNRTI UC-781. The virus strains included the reference strain HIV-1Ba-L and six primary isolates, representative of the HIV-1 group M pandemic. They all display the non-syncytium-inducing and CCR5 receptor-using (NSI/R5) phenotype, important in transmission. Cell-free virus was immobilized on a poly-L-lysine (PLL)-treated microwell plate and incubated with compound for 1 h. Afterwards, the compound was thoroughly washed away; target cells were added and cultured for 2 weeks, followed by an extended culture with highly susceptible mitogen-activated T cells. Viral production in the cultures was measured on supernatant with HIV antigen ELISA. Negative results were confirmed by showing absence of proviral DNA in the cells. TMC-120 and TMC-125 inhibited replication of HIV-1Ba-L with average EC50 values of 38 nM and 117 nM, respectively, whereas the EC50 of UC-781 was 517 nM. Complete suppression of virus and provirus was observed at compound concentrations of 100, 300 and 1000 nM, respectively. Inhibition of all primary isolates followed the same pattern as HIV-1Ba-L. In contrast, pre-treating the virus with the nucleotide RTI PMPA and AZT failed to inhibit infection even at a concentration of 100000 nM. These data clearly suggest that NNRTIs inactivate RT enzymatic activity of different viral clades (predominant in the epidemic) and might be proposed for further testing as a sterilizing microbicide worldwide.
authors.affiliation: Department of Microbiology, Laboratories of Immunology and Virology, Institute of Tropical Medicine, Antwerp, Belgium. hnjai@itg.be
authors.forename: Harr F
authors.initials: HF
authors.lastname: Njai
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Lewi
authors.forename: Cornelus G M
authors.initials: CG
authors.lastname: Janssen
authors.forename: Sergio
authors.initials: S
authors.lastname: Garcia
authors.forename: Katrien
authors.initials: K
authors.lastname: Fransen
authors.forename: Luc
authors.initials: L
authors.lastname: Kestens
authors.forename: Guido
authors.initials: G
authors.lastname: Vanham
authors.forename: Paul A J
authors.initials: PA
authors.lastname: Janssen
mesh_terms: D015496:CD4-Positive T-Lymphocytes; D018920:Coculture Techniques; D003713:Dendritic Cells; D004305:Dose-Response Relationship, Drug; D015658:HIV Infections / Q000821:virology; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects*; D006801:Humans; D007963:Leukocytes, Mononuclear; D009570:Nitriles; D011724:Pyridazines / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D009570:Nitriles; D011724:Pyridazines; D011743:Pyrimidines; C451734:etravirine; D054303:HIV Reverse Transcriptase; C481671:Dapivirine