Journal of Isfahan Medical School

Journal of Isfahan Medical School

Evaluation of the Inhibitory and Destructive Effects of LL-37 Peptide on Biofilms of Uropathogenic Escherichia coli Strains Isolated from Patients with Urinary Tract Infections in Isfahan

Document Type : Original Article(s)

Authors
1 PhD Candidate of Microbiology, Division of Microbiology, Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran
2 PhD of Bacteriology, Division of Microbiology, Associate Professor, Department of Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran
3 PhD of Microbiology, Assistant Professor, Department of Biology, Faculty of Science, University of Sistan and Baluchestan, Zahedan, Iran
10.48305/jims.v44.i864.0868
Abstract
Background: The human cathelicidin antimicrobial peptide (LL-37), a component of the host innate immune system, possesses the capability to combat bacterial biofilms. This study aimed to evaluate the effect of sub-MIC concentrations of LL-37 on biofilms formed by clinical uropathogenic Escherichia coli (UPEC) strains isolated from patients in Isfahan, Iran, during 2024.
Methods: A total of 247 suspected E. coli isolates were collected from patients and identified by standard biochemical tests, and finally confirmed using PCR by specific primers for tufA gene. Biofilm-forming ability was assessed using qualitative Congo Red Agar (CRA) and quantitative Microtiter Plate (MTP) methods, and the frequency of biofilm-associated genes was determined. The minimum inhibitory concentration (MIC) of the LL-37 cathelicidin (ProteoGenix France) was evaluated; and subsequently, the activity of LL-37 at sub-MIC concentrations on biofilm inhibition and destruction among six selected strains (strong, moderate, and weak biofilm-producers) was investigated.
Findings: Of the 204 confirmed strains, 57% were biofilm producers. The yedQ gene was detected in all biofilm-producing strains. The 12 µM concentration of LL-37 reduced biofilm formation in strong and moderate biofilm producers by 34-40% and 41-48%, respectively. Furthermore, this concentration disrupted pre-formed biofilms with an efficacy of 38-53%; however, the 6 µM concentration showed a limited effect (<10%) on weak biofilm producers.
Conclusion: TThe LL-37 peptide exhibits significant anti-biofilm activity at sub-MIC concentrations against clinical UPEC strains, particularly strong biofilm producers. These findings highlight the potential of LL-37 as an adjunctive therapeutic agent in the management of urinary tract infections.

Highlights

Narges Sadat Mostafavi Darani: Google Scholar

Fateh Rahimi: Google Scholar , PubMed

Ali Qasemi: Google Scholar , PubMed

Keywords
Subjects

1.     Tullus K, Shaikh N. Urinary tract infections in children. The Lancet. 2020;395(10237):1659-68.
2.     Rahimi F, Khashei S. Investigation the effect of nitric oxide on biofilm-producing uropathogenic Escherichia coli strains isolated from patients [in Persian]. Tehran Univ Med Sci J 2024; 82(7): 528-37.
3.     Mahale RP, Solomon AP, Maheshwarappa YD, Mahadevaiah S. Comparative evaluation of biofilm-forming capacity in uropathogenic and commensal Escherichia coli. Front Cell Infect Microbiol 2025; 15: 1570422.
4.     Flores-Mireles AL, Walker JN, Caparon M, Hultgren SJ. Urinary tract infections: epidemiology, mechanisms of infection and treatment options. Nat Rev Microbiol 2015; 13(5): 269-84.
5.     Micali S, Isgro G, Bianchi G, Miceli N, Calapai G, Navarra M. Cranberry and recurrent cystitis: more than marketing? Crit Rev Food Sci Nutr 2014; 54(8): 1063-75.
6.     Jhang J-F, Kuo H-C. Recent advances in recurrent urinary tract infection from pathogenesis and biomarkers to prevention. Tzu Chi Medical Journal 2017; 29(3): 131-7.
7.     Soto SM, Marco F, Guiral E, Vila J. Biofilm formation in uropathogenic Escherichia coli strains: Relationship with urovirulence factors and antimicrobial resistance. Clinical Management of Complicated Urinary Tract Infection. 1st ed. London, UK: IntechOpen; 2011.
8.     Oelschlaeger T, Dobrindt U, Hacker J. Pathogenicity islands of uropathogenic E. coli and the evolution of virulence. Int J Antimicrob Agents 2002; 19(6): 517-21.
9.     Qasemi A, Rahimi F, Katouli M. Clonal groups of extended-spectrum β-lactamase and biofilm producing uropathogenic Escherichia coli in Iran [in Persian]. Pathogens and Global Health 2022; 116(8): 485-97.
10.  Qasemi A, Rahimi F, Katouli M. Genetic diversity and virulence characteristics of biofilm-producing uropathogenic Escherichia coli. Int Microbiol 2022; 25(2): 297-307.
11.  Wiles TJ, Kulesus RR, Mulvey MA. Origins and virulence mechanisms of uropathogenic Escherichia coli. Exp Mol Patho 2008; 85(1): 11-9.
12.  Zhao F, Yang H, Bi D, Khaledi A, Qiao M. A systematic review and meta-analysis of antibiotic resistance patterns, and the correlation between biofilm formation with virulence factors in uropathogenic E. coli isolated from urinary tract infections. Microb Pathog 2020; 144: 104196.
13.  Serra DO, Richter AM, Hengge R. Cellulose as an architectural element in spatially structured Escherichia coli biofilms. Microb Pathogc 2013; 195(24): 5540-54.
14.  Verma P, Ho R, Chambers SA, Cegelski L, Zimmer J. Insights into phosphoethanolamine cellulose synthesis and secretion across the Gram-negative cell envelope. Nat Commun 2024; 15(1): 7798.
15.  Lindenberg S, Klauck G, Pesavento C, Klauck E, Hengge R. The EAL domain protein YciR acts as a trigger enzyme in ac‐di‐GMP signalling cascade in E. coli biofilm control. EMBO J 2013; 32(14): 2001-14.
16.  Kai-Larsen Y, Lüthje P, Chromek M, Peters V, Wang X, Holm Å, et al. Uropathogenic Escherichia coli modulates immune responses and its curli fimbriae interact with the antimicrobial peptide LL-37. PLoS Pathog 2010; 6(7): e1001010.
17.  Santos J, Ventura S, Pallarès I. LL-37 and CsgC exemplify the crosstalk between anti-amyloid, antimicrobial, and anti-biofilm protein activities. Neural Regen Res 2023; 18(5): 1027-8.
18.  Chromek M, Slamová Z, Bergman P, Kovács L, Podracka Lu, Ehrén I, et al. The antimicrobial peptide cathelicidin protects the urinary tract against invasive bacterial infection. Nat Med 2006; 12(6): 636-41.
19.  Ridyard KE, Elsawy M, Mattrasingh D, Klein D, Strehmel J, Beaulieu C, et al. Synergy between human peptide LL-37 and polymyxin B against planktonic and biofilm cells of Escherichia coli and Pseudomonas aeruginosa. Antibiotics (Basel) 2023; 12(2): 389.
20.  Pereira AC, Aguiar AP, Araujo LM, Dantas LO, Mayer MP, Karygianni L, et al. Antibiofilm activity of LL-37 peptide and D-amino acids associated with antibiotics used in regenerative endodontics on an ex vivo multispecies biofilm model. Life (Basel) 2022; 12(11): 1686.
White JK, Muhammad T, Alsheim E, Mohanty S, Blasi-Romero A, Gunasekera S, et al. A stable cyclized antimicrobial peptide derived from LL-37 with host immunomodulatory effects and activity against uropathogens. Cell Mol Life Sci 2022; 79(8): 411
Volume 44, Issue 864
4th Week,July
July and August 2026
Pages 868-875

  • Receive Date 27 September 2025
  • Accept Date 26 July 2026