Beta-defensin 118 (Homo sapiens)

Antibiotics are widely used to treat various inflammatory bowel diseases caused by enterotoxigenic Escherichia coli (ETEC). However, continuous use of antibiotics may lead to drug resistance. In this study, we investigated the role of human β-defensin 118 (DEFB118) in regulating the ETEC-induced inflammation and intestinal injury. ETEC-challenged or non-challenged mice were treated by different concentrations of DEFB118. We show that ETEC infection significantly increased fecal score (P < 0.05) and serum concentrations of interlukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Moreover, the concentrations of D-lactic acid, C-reactive protein (CRP), creatinine (CREA), and urea (P < 0.05) were both increased in the ETEC-challenged mice. However, DEFB118 significantly decreased their concentrations in the serum (P < 0.05). DEFB118 not only alleviated tissue damage in spleen upon ETEC challenge, but also increased the villus height in duodenum and ileum (P < 0.05). Moreover, DEFB118 improved the localization and abundance of tight junction protein ZO-1 in jejunal epithelium. Interestingly, DEFB118 decreased the expression levels of critical genes involving in mucosal inflammatory responses (NF-κB, TLR4, IL-1β, and TNF-α) and the apoptosis (caspase3) upon ETEC challenge (P < 0.05), whereas DEFB118 significantly upregulated the expression of mucosa functional genes such as the mucin1 (MUC1) and sodium-glucose transporter-1 (SGLT-1) in the ETEC-challenged mice (P < 0.05). These results indicated a novel function of the DEFB118. The anti-inflammatory effect of DEFB118 should make it an attractive candidate to prevent various bacteria-induced inflammatory bowel diseases.

Purpose: β-Defensin 118 (DEFB118) is a novel host defense peptide (HDP) identified in humans. To evaluate its potentials for future utilization, the DEFB118 gene was expressed in Escherichia coli (E. coli) and the recombinant protein was fully characterized. Methods: The DEFB118 protein was obtained by heterologous expression using E. coli Rosetta (DE3). Antibacterial activity of DEFB118 was determined by using various bacterial strains. IPEC-J cells challenged by E. coli K88 were used to determine its influences on inflammatory responses. Results: The E. coli transformants yielded more than 250 μg/mL DEFB118 protein after 4 h induction by 1.0 mM IPTG. The DEFB118 was estimated by SDS-PAGE to be 30 kDa, and MALDI-TOF analysis verified that it is a human β-defensin 118. Importantly, the DEFB118 showed antimicrobial activities against both Gram-negative bacteria (E. coli K88 and E. coli DH5α) and Gram-positive bacteria (S. aureus and B. subtilis), with a minimum inhibitory concentration (MIC) of 4 μg/mL. Hemolytic assays showed that DEFB118 had no detrimental impact on cell viability. Additionally, DEFB118 was found to elevate the viability of IPEC-J2 cells upon E. coli K88 challenge. Moreover, DEFB118 significantly decreased cell apoptosis in the late apoptosis phase and downregulated the expression of inflammatory cytokines such as IL-1β and TNF-α in IPEC-J2 cell exposure to E. coli K88.

The family of human β-defensins consists of small cysteine-rich peptides, which are receiving significant attention due to their antimicrobial activity. The N-terminal cysteine motif of β-defensin is considered to contribute to its biological activity. Human β-defensin 118 (DEFB 118) is a particular anion β-defensin expressed predominantly in the male reproductive tract, but its physiological activity has not yet been revealed. In order to verify the potential role of the N-terminal domain of DEFB118 peptide in the regulation of infection, the truncated β-defensin core region of DEFB118 peptide was expressed with IMPACT-pTWIN1 system in Escherichia coli. Herein, the purified homogeneous DEFB118 peptide was identified by mass spectrometry and circular dichroism spectroscopy. The in vitro experiments revealed that DEFB118 peptide exhibited prominent LPS-binding potency (KD: 2.94 nM). Moreover, the DEFB118 core peptide significantly inhibited the mRNA level of LPS-induced inflammatory cytokines including IL-α, IL-1β, IL-6 and TNF-α in RAW264.7 cells, and correspondingly decreased secretion of IL-6 and TNF-α. We concluded that strong binding of DEFB118 to LPS might prevent LPS from binding to its receptor, and hence inhibited cytokines secretion. The results of this study may be a benefit to elucidate the immune protection of DEFB118 in the male reproductive tract.