Antimicrobial peptide CAMA‐syn expressed in pulmonary epithelium by recombination adenovirus inhibited the growth of intracellular bacteria

Abstract

Background Intracellular bacteria, especially Mycobacterium tuberculosis, are important pathogenic microorganisms that endanger human health. Purified and synthesized cecropin A‐magainin 2 (CAMA‐syn) can exhibit a higher antibacterial activity and lower cytotoxicity. To enhance such antimicrobial potential, it would be desirable to deliver CAMA‐syn expressed in lung epithelial cells by an adenovirus vector using gene therapy. Methods A549 cells in vitro and lung epithelial cells in vivo were used to express CAMA‐syn by transducing recombinant adenovirus Ad‐SPC‐CAMA/GFP, and the expression of CAMA‐syn was determined by a reverse transcriptase‐polymerase reaction (RT‐PCR) and immunofluorescence. The antimicrobial activity in cells was investigated by colony‐forming rate and growth curve. Forty Kunming mice of a Bacillus Calmette‐Guerin (BCG) infection animal model were randomly divided into three groups: adenoviruses delivery of Ad‐SPC‐CAMA/GFP, Ad‐CMV‐CAMA/GFP and empty‐virus Ad‐CMV‐GFP. The expression of CAMA‐syn in mice was confirmed by RT‐PCR and immunofluorescence. After tracheal injection of adenoviral vector for 3 days, lungs from the mouse model were extracted and homogenized for detection of colony‐forming efficiency. Results CAMA‐syn expressed in lung epithelial cells A549 conferred antimicrobial activity against a series of bacteria, including Salmonella abortusovis and BCG. The results obtained in vivo showed that the colony‐forming rate of Ad‐SPC‐CAMA/GFP (74.54%) and Ad‐CMV‐CAMA/GFP (62.31%) transduced into mice was significantly lower than that of the control group. Conclusions Lung epithelial‐specific expression of antimicrobial peptide CAMA‐syn mediated by adenovirus suppressed the growth of intracellular bacteria, providing a promising approach for the control of refractory intracellular infection.

Galleria mellonella–intracellular bacteria pathogen infection models: the ins and outs

“…Protein expression was dose-dependent, with a higher CFU resulting in a higher differential in protein abundance. Detection of cecropin A was notable as a synthetic hybrid peptide containing the alpha-helical motif of cecropin A has antimycobacterial activity against BCG (Yue et al 2020 ). Furthermore, cecropin A is known to possess immunomodulating activity like the human AMP LL-37, a key component of hCAP-18 treatment for tuberculosis (Torres-Juarez et al 2015 ).…”

Antimicrobial Peptides (AMPs) and the Microbiome in Preterm Infants: Consequences and Opportunities for Future Therapeutics

“…The lung epithelium is rich in immune cells including alveolar macrophages and epithelial cells, which are all able to produce AMPs to protect the lungs from pathogens entering the airways [ 43 , 170 , 171 , 172 , 173 , 174 ]. Most data on preterm and term infants derive from in vitro experiments, studies of fetal lungs and the bronchoalveolar fluid of ventilated neonates.…”

A Meta-analysis of Antimicrobial Peptide Effects on Intestinal Bacteria, Immune Response, and Antioxidant Activity of Broilers

“…is possibly due to the ability of AMP in the form of cecropin-A-maganin-2 (CAMA) to inhibit or even kill gram-positive bacteria (Vizioli et al, 2000). CAMA is composed of an amphipathic terminal base in CA and N-terminal (hydrophobic region) base in MA that both terminals were effective in damaging bacterial cell membranes (Park & Yoe, 2017a;Xiao et al, 2015;Yue et al, 2020;Zhang et al, 2017).…”