A dual chamber model of female cervical mucosa for the study of HIV transmission and for the evaluation of candidate HIV microbicides.
A dual chamber system was established to model heterosexual HIV transmission. Cell-associated, but not cell-free HIV, added to a confluent layer of cervical epithelial cells in the apical chamber, reproducibly infected monocyte-derived dendritic cells (MO-DC) and CD4(+) T cells in the basal compartment. Only minimal epithelial transmigration of HIV-infected mononuclear cells (HIV-PBMCs) was observed. Most evidence points to transepithelial migration of virus, released from HIV-PBMCs after their activation by epithelial cells. We used this model for evaluation of the therapeutic index of various potentially preventive antiviral compounds, including non-nucleoside reverse transcriptase inhibitors (NNRTIs, including UC781 and various diaryltriazines and diarylpyrimidines), poly-anionic entry inhibitors (including PRO2000, cellulose sulphate, dextrane sulphate 5000 and polystyrene sulphonate) and the fusion inhibitor T-20. The epithelium was pre-treated with compound and incubated with HIV-PBMCs for 24 h. Afterwards the apical chamber was removed and MO-DC/CD4(+) T cell co-cultures were further cultured without compound. NNRTIs, including a TMC120 gel, blocked infection of the sub-epithelial targets at sub-micromolar concentrations. Polyanionic entry inhibitors (up to 100 microg/ml) and T-20 (up to 449 microg/ml) failed to inhibit transmission. Moreover, whereas the NNRTIs used interfered with epithelial integrity with cervical epithelium only at very high concentrations, the evaluated entry inhibitors showed toxicity at concentrations that did not prevent infection.
Van Herrewege
Yven
Y
Virology Unit, Department of Microbiology, Institute of Tropical Medicine, 155 Nationalestraat, B-2000 Antwerpen, Belgium. yvherrewege@itg.be
Michiels
Jo
J
Waeytens
Anouk
A
De Boeck
Gitte
G
Salden
Evelyne
E
Heyndrickx
Leo
L
van den Mooter
Guy
G
de Béthune
Marie-Pierre
MP
Andries
Koen
K
Lewi
Paul
P
Praet
Marleen
M
Vanham
Guido
G
D019380:Anti-HIV Agents / Q000494:pharmacology*; D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000821:virology*; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000821:virology; D004847:Epithelial Cells / Q000821:virology; D005260:Female; D005434:Flow Cytometry; D015658:HIV Infections / Q000517:prevention & control / Q000635:transmission*; D015497:HIV-1 / Q000187:drug effects* / Q000254:growth & development; D006801:Humans; D018613:Microscopy, Confocal; D009092:Mucous Membrane / Q000821:virology
D019380:Anti-HIV Agents