title: Utilization of plant-derived recombinant human β-defensins (hBD-1 and hBD-2) for averting salmonellosis.
abstract: We describe the use of plant-made β-defensins as effective antimicrobial substances for controlling salmonellosis, a deadly infection caused by Salmonella typhimurium (referred to further as S. typhi). Human β-defensin-1 (hBD-1) and -2 (hBD-2) were expressed under the control of strong constitutive promoters in tobacco plants, and bio-active β-defensins were successfully extracted. In the in vitro studies, enriched recombinant plant-derived human β-defensin-1 (phBD-1) and -2 (phBD-2) obtained from both T1 and T2 transgenic plants showed significant antimicrobial activity against Escherichia coli and S. typhi when used individually and in various combinations. The 2:1 peptide combination of phBD-1:phBD-2 with peptides isolated from T1-and T2-generation plants reduced the growth of S. typhi by 96 and 85 %, respectively. In vivo studies employing the mouse model (Balb/c) of Salmonella infection clearly demonstrated that the administration of plant-derived defensins individually and in different combinations enhanced the mean survival time of Salmonella-infected animals. When treatment consisted of the 2:1 phBD-1:phBD-2 combination, approximately 50 % of the infected mice were still alive at 206 h post-inoculation; the lowest number of viable S. typhi was observed in the liver and spleen of infected animals. We conclude that plant-made recombinant β-defensins (phBD-1 and phBD-2) are promising antimicrobial substances and have the potential to become additional tools against salmonellosis, particularly when used in combination.
authors.affiliation: Division of Gene Function and Regulation, Department of Biotechnology, Institute of Life Sciences, Govt. of India, Nalco Square, Chandrasekherpur, Bhubaneswar, 751 023, Odisha, India.
authors.forename: Sunita
authors.initials: S
authors.lastname: Patro
authors.forename: Soumitra
authors.initials: S
authors.lastname: Maiti
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Panda
authors.forename: Nrisingha
authors.initials: N
authors.lastname: Dey
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D051379:Mice; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D012475:Salmonella / Q000187:drug effects / Q000472:pathogenicity; D012480:Salmonella Infections / Q000188:drug therapy* / Q000382:microbiology; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D023083:beta-Defensins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics
chemical_list: C406407:DEFB1 protein, human; C106641:DEFB4A protein, human; D012333:RNA, Messenger; D023083:beta-Defensins