Anti-heparan Sulfate Peptides That Block Herpes Simplex Virus Infection in Vivo

• DOI: 10.1074/jbc.m110.201103
• Publication Date: 2011-05-20T00:47:07Z
• Authors: Vaibhav Tiwari; Jian Liu; Tibor Valyi-Nagy; Deepak Shukla
• Keywords: 0301 basic medicine; Mice, Inbred BALB C; Eye Infections, Viral; Herpes Simplex; CHO Cells; Herpesvirus 1, Human; Virus Internalization; Antiviral Agents; 3. Good health; Mice; 03 medical and health sciences; Cricetulus; Amino Acid Substitution; Viral Envelope Proteins; Cricetinae; Chlorocebus aethiops; Animals; Humans; Point Mutation; Heparitin Sulfate; Peptides; Vero Cells; HeLa Cells

Heparan sulfate (HS) and its highly modified form, 3-O-sulfated heparan sulfate (3-OS HS), contribute strongly to herpes simplex virus type-1 (HSV-1) infection in vitro. Here we report results from a random M13-phage display library screening to isolate 12-mer peptides that bind specifically to HS, 3-OS HS, and block HSV-1 entry. The screening identified representative candidates from two-different groups of anti-HS peptides with high positive charge densities. Group 1, represented by G1 peptide (LRSRTKIIRIRH), belongs to a class with alternating charges (XRXRXKXXRXRX), and group 2, represented by G2 peptide (MPRRRRIRRRQK), shows repetitive charges (XXRRRRXRRRXK). Viral entry and glycoprotein D binding assays together with fluorescent microscopy data indicated that both G1 and G2 were potent in blocking HSV-1 entry into primary cultures of human corneal fibroblasts and CHO-K1 cells transiently expressing different glycoprotein D receptors. Interestingly, G2 peptide isolated against 3-OS HS displayed wider ability to inhibit entry of clinically relevant strains of HSV-1 and some divergent members of herpesvirus family including cytomegalovirus and human herpesvirus-8. To identify functional residues within G1 and G2, we performed point mutations and alanine-scanning mutagenesis. Several arginine and a lysine residues were needed for anti-HSV-1 activity, suggesting the importance of the positively charged residues in virus-cell binding and virus-induced membrane fusion. In vivo administration of G1 or G2 peptide as a prophylactic eye drop completely blocked HSV-1 spread in the mouse cornea as evident by immunohistochemistry. This result also highlights an in vivo significance of HS and 3-OS HS during ocular herpes infection.