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.
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Acknowledgments
We are grateful to the Council for Scientific and Industrial Research, Govt. of India for financial support [Project No. 38(1268)/10/EMR-II to ND], Fellowship (20-6/2008 (ii) EU-IV) to SP and ILS/Core fund to ND. We sincerely thank the Director, Institute of Life Sciences, Bhubaneswar, for his constant inspiration and suggestion for pursuing this study. We are grateful to Prof. T. Ganz for providing the cDNA clones of hBD-1 and hBD-2 and to Dr. D.V Singh for providing the bacterial strains. We also thank Mr. Sukumar Purohit for his kind help in performing the mice experiments and Ms. Vineeta Rai for providing the transformed control plant used in this study. The technical support provided by Mr. Abhimanyu Das is gratefully acknowledged.
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Sunita Patro and Soumitra Maiti have contributed equally to this work.
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Patro, S., Maiti, S., Panda, S.K. et al. Utilization of plant-derived recombinant human β-defensins (hBD-1 and hBD-2) for averting salmonellosis. Transgenic Res 24, 353–364 (2015). https://doi.org/10.1007/s11248-014-9847-3
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DOI: https://doi.org/10.1007/s11248-014-9847-3