Viral replication after ex vivo infection with HSV-2
HSV-2
Two ectocervical biopsies were obtained at baseline and 24 hours after insert dosing. Ectocervical biopsies were placed into 1ml of explant media RPMI 1640 (Gibco, 11875093) 10%FBS, 100u/ml Pen/Strep (Gibco, 15140122) in the clinic. The biopsies were infected with 4X10To perform the PCR the supernatants were thawed on ice. The PCR kit was purchased from Roche (catalogue number: 12239264001) and was used according to manufacturer’s protocol. HSV-2 DNA was evaluated by a quantitative PCR amplification of tissue culture supernatants using SYBR-green. Supernatants (6 μl) were amplified using the forward primer 5′- TCGCCAGCACAAACTCAT -3′ and the reverse primer 5′- CCACCGACCTCAAGTACAAC-3′ targeting glyprotein B. All data were normalized to day 0.