Antibacterial activity of a synthetic peptide that mimics the LPS binding domain of Indian mud crab, Scylla serrata Anti-lipopolysaccharide Factor (SsALF) also involved in the modulation of vaginal immune functions through NF-kB signaling

A Rabbit Vaginal Cell-Derived Antimicrobial Peptide, RVFHbαP, Blocks Lipopolysaccharide-Mediated Inflammation in Human Vaginal CellsIn Vitro

“…HeLa hVECs used in this study constitutively express TLR4 and respond to LPS (27,33). On the day of treatment, cells at 70 to 80% confluence were split by trypsin treatment (0.1%).…”

“…After the treatment, hVECs were lysed with hypotonic HEPES lysis buffer (pH 7.4) and centrifuged at 1,000 ϫ g for 10 min at 4°C. Supernatants were collected, and total protein levels were determined (4) and used for the estimation of TLR4 levels by ELISA as described earlier (33).…”

“…The treatment details are the same as those described above for NF-B Western blot analysis. After treatment, hVECs were lysed with hypotonic HEPES lysis buffer (pH 7.4) and centrifuged at 1,000 ϫ g for 10 min at 4°C, and supernatants were collected and used for the estimation of NF-B p65 as described earlier (33).…”

“…The vagina is a major portal of entry for the pathogens associated with sexually transmitted infections (STIs), such as those from Neisseria gonorrhoeae, Chlamydia trachomatis, Haemophilus ducreyi, Treponema pallidum, and viruses like human immunodeficiency virus type 1 (HIV-1), herpes simplex virus 1 (HSV-1), and human papillomavirus (HPV), which represent major threats to human health (29,33). In a large part of bacterial infections, products like lipopolysaccharide (LPS) from Gram-negative bacteria (e.g., Escherichia coli) and lipoteichoic acid (LTA) from Gram-positive bacteria (e.g., Staphylococcus aureus) are involved (5,43).…”

“…Therefore, identifying the molecule(s) which binds to LPS and neutralizes its activities might have important clinical applications (25,33). In the past, to prevent LPS-induced inflammatory events in the host, several approaches have been employed (23), including specific antibodies (Abs) directed against LPS.…”