Targeting expression of antimicrobial peptide CAMA-Syn by adenovirus vector in macrophages inhibits the growth of intracellular bacteria

Zhang, Junlin; Xie, Lin; Xu, Dake; Yue, Shuohao; Li, Yi; Guo, Xiaohong; Lai, Xiaojing

doi:10.1016/j.gene.2017.07.079

Cited by 8 publications

(8 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We had previously reported the application of macrophage specific promoter in lymphocyte to express the antimicrobial peptide CAMA-syn mediated by adenovirus. 9 The results obatained showed that the adenovirus expression system targeting macrophages ; all experiments were repeated three times demonstrated that the Ad-SPC-GFP/CAMA and Ad-CMV-CAMA/GFP showed better antibacterial activity with an inhibition rate of 24.4-38.4% to BCG, which was different compared to S. aureus, P. aeruginosa and E. coli. Our results have provided a promising approach for the expression of AMPs in the control of refractory infection, via the expression of CAMA-syn in lung epithelial cells by an adenovirus vector.…”

Section: Discussionmentioning

confidence: 92%

“…We had previously reported the application of macrophage specific promoter in lymphocyte to express the antimicrobial peptide CAMA‐syn mediated by adenovirus . The results obatained showed that the adenovirus expression system targeting macrophages inhibited both Gram‐positive and Gram‐negative bacteria in vitro and in vivo , and also demonstrated certain antimicrobial activity in a BCG infection model of the mouse.…”

Section: Discussionmentioning

confidence: 98%

“…The adenovirus control vector that lacks the CAMA‐syn gene was named as Ad‐CMV‐GFP. Recombinant adenoviruses were packaged in 293A cells and virus titers were measured by plaque assays as described in our previous research …”

Section: Methodsmentioning

confidence: 99%

“…Mice were anesthetized with 0.1 mL of 2% pentobarbital sodium by retroperitoneal injection. Tracheal injection was inducted as described previously . Next, 100 μL of rAd (1 × 10 9 pfu) was injected into trachea.…”

Section: Methodsmentioning

confidence: 99%

“…Tracheal injection was inducted as described previously. 9 Next, 100 μL of rAd (1 × 10 9 pfu) was injected into trachea. Intravenous infusion and retroperitoneal infusion were performed in accordance with standard operating procedures.…”

Section: Recombinant Adenovirus Injection Methodsmentioning

confidence: 99%

See 4 more Smart Citations

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

Yue

Jie

Xie

et al. 2020

The Journal of Gene Medicine

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Recombinant Adenovirus Injection Methodsmentioning

confidence: 99%

See 3 more Smart Citations