Cyclosporine

All patients received induction treatment with a regimen composed of antithymocyte globulin (ATG, Thymoglobulin ®Genzyme Corporation, Cambridge, MA) within 12 h before HTx and continued for up to 3 days. Patients were randomized within 5 days to either the EVR or the CNI group. The EVR group was treated with low‐exposure EVR (Certican; Novartis Pharma AG, Basel, Switzerland), low‐exposure cyclosporine (Sandimmun Neoral; Novartis Pharma AG), MMF, and CSs with cyclosporine withdrawal between 7 and 11 weeks post‐HTx. The CNI group was treated with cyclosporine, MMF, and CSs. Target trough levels and target doses of the treatment have been previously reported.19

Four-week-old juvenile wild-type ferrets (n = 15 per experimental group) were anesthetized with a mixture of ketamine and xylazine injected subcutaneously and were i.t. administered AAV2.5T-SP183-fCFTRΔR at a dose of 1 × 10¹³ DRP/kg body weight (BW). Four weeks later, anesthetized ferrets were administered 1 × 10¹³ DRP/kg BW AAV2.5T-SP183-gLuc. The ferrets were dosed using a microsprayer aerosolizer (PennCentury, model IA-1B). The AAV2.5T vector inoculum contained doxorubicin at a concentration of 200 μM, and the administered volume was normalized to ferret BW at 2 mL/kg BW.
Daily administration of immunosuppressants started 2 days before the first vector dose and continued until the dosing day of the second vector dose (30 days of total dosing). Specifically, the cyclosporine (1 mg/kg BW, Novartis) and MP (2 mg/kg BW, Pfizer) were administered intraperitoneally, and azathioprine (2 mg/kg BW, Wedgewood Pharmacy) was given orally. The antibiotic Baytril (10 mg/kg BW, Bayer) was injected daily subcutaneously to prevent bacterial infection in the immunosuppressed ferrets.

The HCV NS5A inhibitor daclatasvir (Bristol Myers Squibb), the HCV NS5B polymerase inhibitor sofosbuvir (Gilead), the HCV NS3 protease inhibitors boceprevir (Merck) and telaprevir (Vertex) and the HIV-1 reverse transcriptase inhibitor emtricitabine (Gilead) were all obtained from MedChemexpress (Princeton, NJ 08540, USA). Alisporivir and NIM811 were generously provided by Novartis, whereas cyclosporine A, sanglifehrins A and B were generously provided by Drs. Wilkinson and Gregory. Poly I:C was obtained from InvivoGen (San Diego, CA, USA).