Expression of T9W in Pichia pastoris and the protective roles of T9W in ICR Mice.

BACKGROUND: The purpose of this paper is to express the novel α-helical peptide T9W more efficiently using the Pichia pastoris expression system and to examine the role of T9W in ICR mice. RESULTS: The novel antimicrobial peptide T9W was expressed in P. pastoris X-33 by using the vector pPICZαA. Approximately 13 mg/L T9W was secreted from the culture and purified. The expressed peptide has similar activity to the synthetic peptide. ICR female mice challenged with P. aeruginosa 27853 at the LD100 were treated with T9W and CPFX. The results showed that the secretion of inflammatory cytokines and lung damage was significantly reduced by the treatment group, and the protective response was equivalent between T9W and ciprofloxacin-treated mice. CONCLUSION: T9W was expressed in P. pastoris X-33 via the methanol-inducible vector pPICZαA and exhibited the same biological activity as synthetic T9W. T9W can alleviate damage to mice caused by P. aeruginosa.

TLR2/4-mediated NF-κB pathway combined with the histone modification regulates ß-defensins and interleukins expression by sodium phenyl butyrate in porcine intestinal epithelial cells.

BACKGROUND: Host defense peptides (HDPs) possess direct antibacterial, antineoplastic, and immunomodulatory abilities, playing a vital role in innate immunity. Dietary-regulated HDP holds immense potential as a novel pathway for preventing infection. OBJECTIVE: In this study, we examined the regulation mechanism of HDPs (pEP2C, pBD-1, and pBD-3) and cytokines (IL-8 and IL-18) expression by sodium phenylbutyrate (PBA). DESIGN: The effects of PBA on HDP induction and the mechanism involved were studied in porcine intestinal epithelial cell lines (IPEC J2). RESULTS: In this study, the results showed that HDPs (pEP2C, pBD-1, and pBD-3) and cytokines (IL-8 and IL-18) expression was increased significantly upon stimulation with PBA in IPEC J2 cells. Furthermore, toll-like receptor 2 (TLR2) and TLR4 were required for the PBA-mediated upregulation of the HDPs. This process occurred and further activated the NF-κB pathway via the phosphorylation of p65 and an IκB α synthesis delay. Meanwhile, histone deacetylase (HDAC) inhibition and an increased phosphorylation of histone H3 on serine S10 also occurred in PBA-induced HDP expression independently with TLR2 and TLR4. Furthermore, p38-MAPK suppressed PBA-induced pEP2C, pBD-1 pBD-3, IL-8, and IL-18 expression, but ERK1/2 failed to abolish the regulation of pBD-3, IL-8, and IL-18. Moreover, epidermal growth factor receptor (EGFR) is involved in PBA-mediated HDP regulation. CONCLUSIONS: We concluded that PBA induced HDP and cytokine increases but did not cause an excessive pro-inflammatory response, which proceeded through the TLR2 and TLR4-NF-κB pathway and histone modification in IPEC J2 cells.