Histidine-rich Modification of a Scorpion-derived Peptide Improves Bioavailability and Inhibitory Activity against HSV-1

Zeng, Zhengyang; Zhang, Runhong; Hong, Wei; Cheng, Yuting; Wang, Huijuan; Lang, Yange; Ji, Zhenglin; Wu, Yingliang; Li, Wenxin; Xie, Youli; Cao, Zhijian

African green monkey kidney cells (Vero) were cultured at 37 ℃ and 5% CO 2 in minimum essential medium (MEM) (Invitrogen, Foster, CA, USA) containing 10% (vol/vol) fetal bovine serum (FBS) (Gibco, Foster, CA, USA), 100 U/mL penicillin and 100 μg/mL streptomycin (Sigma, St. Louis, MO, USA). Cells infected with HSV-1 were cultured in MEM with 2% FBS in the same environment as described above. High-titer stocks of HSV-1 (F strain) were prepared as follows. The original viral stocks were diluted in MEM and inoculated into 80%-90% confluent Vero cell cultures at a multiplicity of infection (MOI) of 0.1 in a T25 flask (Corning, New York, USA). When the cytopathic effect (CPE) reached 90%-100%, the infected monolayers were harvested and subjected to three freeze-thaw cycles using a dry ice-ethanol bath and then centrifuged at 1000 × g for 5 min. The supernatant was collected after centrifugation. After titration, the virus aliquots were stored at -80 ℃.