Abstract
Uropathogenic Escherichia coli (UPEC) strains are the primary cause of urinary tract infections (UTIs). UPEC strains are able to invade, multiply and persisting in host cells. Therefore, UPEC strains are associated to recurrent UTIs requiring long‐term antibiotic therapy. However, this therapy is suboptimal due to the increase of multidrug-resistant UPEC. The use of non-antibiotic treatments for managing UTIs is required. Among these, bovine lactoferrin (bLf), a multifunctional cationic glycoprotein, could be a promising tool because inhibits the entry into the host cells of several intracellular bacteria. Here, we demonstrate that 100 μg/ml bLf hinders the invasion of 2.0 ± 0.5 × 104 CFU/ml E. coli CFT073, prototype of UPEC, infecting 2.0 ± 0.5 × 105 cells/ml urinary bladder T24 epithelial cells. The highest protection (100%) is due to the bLf binding with host surface components even if an additional binding to bacterial surface components cannot be excluded. Of note, in the absence of bLf, UPEC survives and multiplies, while bLf significantly decreases bacterial intracellular survival. After these encouraging results, an observational survey on thirty-three patients affected by recurrent cystitis was performed. The treatment consisted in the oral administration of bLf alone or in combination with antibiotics and/or probiotics. After the observation period, a marked reduction of cystitis episodes was observed (p < 0.001) in all patients compared to the episodes occurred during the 6 months preceding the bLf-treatment. Twenty-nine patients did not report cystitis episodes (87.9%) whereas the remaining four (12.1%) experienced only one episode, indicating that bLf could be a worthwhile and safe treatment in counteracting recurrent cystitis.
Similar content being viewed by others
References
Ajello M, Greco R, Giansanti F, Massucci MT, Antonini G, Valenti P (2002) Anti-invasive activity of bovine lactoferrin towards group A streptococci. Biochem Cell Biol 80:119–124. https://doi.org/10.1139/o01-211
Al-Badr A, Al-Shaikh G (2013) Recurrent urinary tract infections management in women: a review. Sultan Qaboos Univ Med J 13:359–367
Alugupalli KR, Kalfas S (1997) Characterization of the lactoferrin-dependent inhibition of the adhesion of Actinobacilllus Actinomycetemcomitans, Prevotella Intermedia and Prevotella Nigrescens to fibroblasts and to a reconstituted basement membrane. APMIS 105:680–688. https://doi.org/10.1111/j.1699-0463.1997.tb05071.x
Antonini G, Catania MR, Greco R, Longhi C, Pisciotta MG, Seganti L, Valenti P (1997) Anti-invasive activity of bovine lactoferrin against listeria monocytogenes. J Food Prot 60(3):267–271. https://doi.org/10.4315/0362-028X-60.3.267
Appelmelk BJ, An YQ, Geerts M, Thijs BG, de Boer HA, MacLaren DM, de Graaff J, Nuijens JH (1994) Lactoferrin is a lipid A-binding protein. Infect Immun 62(6):2628–2632. https://doi.org/10.1128/iai.62.6.2628-2632.1994
Arao S, Matsuura S, Nonomura M, Miki K, Kabasawa K, Nakanishi H (1999) Measurement of urinary lactoferrin as a marker of urinary tract infection. J Clin Microbiol 37(3):553–557. https://doi.org/10.1128/JCM.37.3.553-557.1999
Ashida K, Sasaki H, Sasaki YA, Loönnerdal B (2004) Cellular internalization of lactoferrin in intestinal epithelial cells. Biometals 17:311–315. https://doi.org/10.1023/b:biom.0000027710.13543.3f
Bauckman KA, Matsuda R, Higgins CB, DeBosch BJ, Wang C, Mysorekar IU (2019) Dietary restriction of iron availability attenuates UPEC pathogenesis in a mouse model of urinary tract infection. Am J Physiol Renal Physiol 316(5):F814–F822. https://doi.org/10.1152/ajprenal.00133.2018
Bjerrum L, Gahrn-Hansen B, Grinsted P (2009) Pivmecillinam versus sulfamethizole for short-term treatment of uncomplicated acute cystitis in general practice: a randomized controlled trial. Scand J Prim Health Care 2(1):6–11. https://doi.org/10.1080/02813430802535312
Blango MG, Mulvey MA (2010) Persistence of uropathogenic Escherichia coli in the face of multiple antibiotics. Antimicrob Agents Chemother 54:1855–1863. https://doi.org/10.1128/AAC.00014-10
Bonkat G, Pickard R, Bartoletti R, Cai T, Geerlings SE, Köves B, Schubert S, Pilatz A, Veeratterapillay R, Wagenlehner F (2022) European Association of urology guidelines on urological infections. EAU Guidelines edn. Presented at the EAU annual congress Amsterdam, the Netherlands 2022. ISBN 978-94-92671-16-5. http://uroweb.org/guideline/urological-infections. Accessed 06 Apr 2022.
Brandenburg K, Jürgens G, Müller M, Fukuoka S, Koch MHJ (2001) Biophysical characterization of lipopolysaccharide and lipid a inactivation by lactoferrin. Biol Chem 382:1215–1225. https://doi.org/10.1515/BC.2001.152
Chen L, Xiong Z, Sun L, Yang J, Jin Q (2012) VFDB 2012 update: toward the genetic diversity and molecular evolution of bacterial virulence factors. Nucleic Acids Res 40:D641–D645. https://doi.org/10.1093/nar/gkr989
Croxen MA, Law RJ, Scholz R, Keeney KM, Wlodarska M, Finlay BB (2013) Recent advances in understanding enteric pathogenic Escherichia coli. Clin Microbiol Rev 26(4):822–880. https://doi.org/10.1128/CMR.00022-13
Cutone A, Rosa L, Lepanto MS, Scotti MJ, Berlutti F, Bonaccorsi di Patti MC, Musci G, Valenti P (2017) Lactoferrin efficiently counteracts the inflammation-induced changes of the iron homeostasis system in macrophages. Front Immunol 8:705. https://doi.org/10.3389/fimmu.2017.00705
Cutone A, Lepanto MS, Rosa L, Scotti MJ, Rossi A, Ranucci S, De Fino I, Bragonzi A, Valenti P, Musci G, Berlutti F (2019) Aerosolized bovine lactoferrin counteracts infection, inflammation and iron dysbalance in a cystic fibrosis mouse model of pseudomonas aeruginosa chronic lung infection. Int J Mol Sci 20(9):2128. https://doi.org/10.3390/ijms20092128
Diarra MS, Petitclerc D, Deschenes E, Lessard N, Grondin G, Talbot BG, Lacasse P (2003) Lactoferrin against Staphylococcus aureus mastitis. lactoferrin alone or in combination with penicillin G on bovine polymorphonuclear function and mammary epithelial cells colonisation by Staphylococcus aureus. Vet Immunol Immunopathol 95:33–42. https://doi.org/10.1016/s0165-2427(03)00098-9
Di Biase AM, Tinari A, Pietrantoni A, Antonini G, Valenti P, Conte MP, Superti F (2004) Effect of bovine lactoferricin on enteropathogenic Yersinia adhesion and invasion in HEp-2 cells. J Med Microbiol 53:407–412. https://doi.org/10.1099/jmm.0.05410-0
Elass E, Masson M, Mazurier J, Legrand D (2002) Lactoferrin inhibits the lipopolysaccharide-induced expression and proteoglycan binding ability of interleukin-8 in human endothelial cells. Infect Immun 70:1860–1866. https://doi.org/10.1128/IAI.70.4.1860-1866.2002
European Food Safety Authority (2012) Scientific opinion on bovine lactoferrin. EFSA J 10:2701. https://doi.org/10.2903/j.efsa.2012.2701
Flores-Mireles AL, Walker JN, Caparon M, Hultgren SJ (2015) Urinary tract infections: epidemiology, mechanisms of infection and treatment options. Nat Rev Microbiol 13:269–284. https://doi.org/10.1038/nrmicro3432
Foxman B (2003) Epidemiology of urinary tract infections: incidence, morbidity, and economic costs. Dis Mon 49:53–70. https://doi.org/10.1067/mda.2003.7
Foxman B (2014) Urinary tract infection syndromes: occurrence, recurrence, bacteriology, risk factors, and disease burden. Infect Dis Clin North Am 28:1–13. https://doi.org/10.1016/j.idc.2013.09.003
Frioni A, Conte MP, Cutone A, Longhi C, Musci G, di Patti MC, Natalizi T, Marazzato M, Lepanto MS, Puddu P, Paesano R et al (2014) Lactoferrin differently modulates the inflammatory response in epithelial models mimicking human inflammatory and infectious diseases. Biometals 27(5):843–856. https://doi.org/10.1007/s10534-014-9740-9
Guglietta A (2017) Recurrent urinary tract infections in women: risk factors, etiology, pathogenesis and prophylaxis. Future Microbiol 12:239–246. https://doi.org/10.2217/fmb-2016-0145
Gupta K, Hooton TM, Stamm WE (2001) Increasing antimicrobial resistance and the management of uncomplicated community-acquired urinary tract infections. Ann Intern Med 135(1):41–50. https://doi.org/10.7326/0003-4819-135-1-200107030-00012
Han Z, Pinkner JS, Ford B, Chorell E, Crowley JM, Cusumano CK, Campbell S, Henderson JP, Hultgren SJ, Janetka JW (2012) Lead optimization studies on FimH antagonists: discovery of potent and orally bioavailable ortho-substituted biphenyl mannosides. J Med Chem 55(8):3945–3959. https://doi.org/10.1021/jm300165m
Harding GK, Ronald AR (1994) The management of urinary infections: what have we learned in the past decade? Int J Antimicrob Agents 4:83–88. https://doi.org/10.1016/0924-8579(94)90038-8
Hickling DR, Nitti VW (2013) Management of recurrent urinary tract infections in healthy adult women. Rev Urol 15:41–48. https://doi.org/10.3909/riu0566
Hu Y, Meng X, Zhang F, Xiang Y, Wang J (2021) The in vitro antiviral activity of lactoferrin against common human coronaviruses and SARS-CoV-2 is mediated by targeting the heparan sulfate co-receptor. Emerg Microbes Infect 10(1):317–330. https://doi.org/10.1080/22221751.2021.1888660
Jepson RG, Williams G, Craig JC (2012) Cranberries for preventing urinary tract infections. Cochrane Database Syst Rev 10:CD001321. https://doi.org/10.1002/14651858.CD001321.pub5
Johnson JR (1991) Virulence factors in Escherichia coli urinary tract infection. Clin Microbiol Rev 4:80–128. https://doi.org/10.1128/CMR.4.1.80
Johnson JR, Russo TA (2002) Extraintestinal pathogenic Escherichia coli: “the other bad E. coli". J Lab Clin Med 139(3):155–162. https://doi.org/10.1067/mlc.2002.121550
Johnson JR, Russo TA (2018) Acute pyelonephritis in adults. N Engl J Med 378(1):48–59. https://doi.org/10.1056/nejmcp1702758
Josephs-Spaulding J, Krogh TJ, Rettig HC, Lyng M, Chkonia M, Waschina S, Graspeuntner S, Rupp J, Møller-Jensen J, Kaleta C (2021) Recurrent urinary tract infections: unraveling the complicated environment of uncomplicated rUTIs. Front Cell Infect Microbiol 11:562525. https://doi.org/10.3389/fcimb.2021.562525
Juthani-Mehta M, Van Ness PH, Bianco L, Rink A, Rubeck S, Ginter S, Argraves S, Charpentier P, Acampora D, Trentalange M, Quagliarello V, Peduzzi P (2016) Effect of cranberry capsules on bacteriuria plus pyuria among older women in nursing homes: a randomized clinical trial. JAMA 316(18):1879–1887. https://doi.org/10.1001/jama.2016.16141
Kawasaki Y, Tazume S, Shimizu K, Matsuzawa H, Dosako S, Isoda H, Tsukiji M, Fujimura R, Muranaka Y, Isihida H (2000) Inhibitory effects of bovine lactoferrin on the adherence of enterotoxigenic Escherichia Coli to host cells. Biosci Biotech Biochem 64:348–354. https://doi.org/10.1271/bbb.64.348
Kim CW, Lee TH, Park KH, Choi SY, Kim J (2012) Human lactoferrin suppresses TNF-α-induced intercellular adhesion molecule-1 expression via competition with NF-kB in endothelial cells. FEBS Lett 586:229–234. https://doi.org/10.1016/j.febslet.2011.12.011
Kıraç FS (2013) Is ethics approval necessary for all trials? A clear but not certain process. Mol Imaging Radionucl Ther 22:73–75. https://doi.org/10.4274/Mirt.80664
Kӧhler CD, Dobrindt U (2011) What defines extraintestinal pathogenic Escherichia coli? Int J Med Microbiol 301:642–647. https://doi.org/10.1016/j.ijmm.2011.09.006
Kolman KB (2019) Cystitis and pyelonephritis: diagnosis, treatment, and prevention. Prim Care 46(2):191–202. https://doi.org/10.1016/j.pop.2019.01.001
Lepanto MS, Rosa L, Cutone A, Conte MP, Paesano R, Valenti P (2018) Efficacy of lactoferrin oral administration in the treatment of anemia and anemia of inflammation in pregnant and non-pregnant women: an interventional study. Front Immunol 9:2123. https://doi.org/10.3389/fimmu.2018.02123
Lepanto MS, Rosa L, Paesano R, Valenti P, Cutone A (2019a) Lactoferrin in aseptic and septic inflammation. Molecules 3:24. https://doi.org/10.3390/molecules24071323
Lepanto MS, Rosa L, Cutone A, Scotti MJ, Conte AL, Marazzato M, Zagaglia C, Longhi C, Berlutti F, Musci G, Valenti P, Conte MP (2019b) Bovine lactoferrin pre-treatment induces intracellular killing of AIEC LF82 and reduces bacteria-induced DNA damage in differentiated human enterocytes. Int J Mol Sci 20(22):5666. https://doi.org/10.3390/ijms20225666
Longhi C, Conte MP, Seganti L, Polidoro M, Alfsen A, Valenti P (1993) Influence of lactoferrin on the entry process of Escherichia Coli HB101(PRI203) in HeLa cells. Med Microbiol Immunol 182(1):25–35. https://doi.org/10.1007/BF00195948
Lu J, Francis JD, Guevara MA, Moore RE, Chambers SA, Doster RS, Eastman AJ, Rogers LM, Noble KN, Manning SD, Damo SM, Aronoff DM, Townsend SD, Gaddy JA (2021) Antibacterial and anti-biofilm activity of the human breast milk glycoprotein lactoferrin against group B Streptococcus. ChemBioChem 22(12):2124–2133. https://doi.org/10.1002/cbic.202100016
Minardi D, d’Anzeo G, Cantoro D, Conti A, Muzzonigro G (2011) Urinary tract infections in women: etiology and treatment options. Int J Gen Med 4:333–343. https://doi.org/10.2147/IJGM.S11767
Nicolle LE, Bradley S, Colgan R, Rice JC, Schaeffer A, Hooton TM, Infectious Diseases Society of America, American Society of Nephrology, American Geriatric Society (2005) Infectious Diseases Society of America guidelines for the diagnosis and treatment of asymptomatic bacteriuria in adults. Clin Infect Dis 40:643–654. https://doi.org/10.1086/427507
O’Brien VP, Hannan TJ, Schaeffer AJ, Hultgren SJ (2015) Are you experienced? Understanding bladder innate immunity in the context of recurrent urinary tract infection. Curr Opin Infect Dis 28:97–105. https://doi.org/10.1097/QCO.0000000000000130
O’Brien VP, Hannan TJ, Nielsen HV, Hultgren SJ (2016) Drug and vaccine development for the treatment and prevention of urinary tract infections. Microbiol Spectr. https://doi.org/10.1128/microbiolspec.UTI-0013-2012
Paesano R, Natalizi T, Berlutti F, Valenti P (2012) Body iron delocalization: the serious drawback in iron disorders in both developing and developed countries. Pathog Glob Health 106:200–216. https://doi.org/10.1179/2047773212Y.0000000043
Patras KA, Ha AD, Rooholfada E, Olson J, Ramachandra Rao SP, Lin AE, Nizet V (2019) Augmentation of urinary lactoferrin enhances host innate immune clearance of uropathogenic Escherichia coli. J Innate Immun 11(6):481–495. https://doi.org/10.1159/000499342
Phé V, Pakzad M, Haslam C, Gonzales G, Curtis C, Porter B, Chataway J, Panicker JN (2017) Open label feasibility study evaluating D-mannose combined with home-based monitoring of suspected urinary tract infections in patients with multiple sclerosis. Neurourol Urodyn 36:1770–1775. https://doi.org/10.1002/nau.23173
Rosa L, Cutone A, Lepanto MS, Paesano R, Valenti P (2017) Lactoferrin: a natural glycoprotein involved in iron and inflammatory homeostasis. Int J Mol Sci 18:1985. https://doi.org/10.3390/ijms18091985
Rosa L, Lepanto MS, Cutone A, Siciliano RA, Paesano R, Costi R, Musci G, Valenti P (2020) Influence of oral administration mode on the efficacy of commercial bovine Lactoferrin against iron and inflammatory homeostasis disorders. Biometals 33(2–3):159–168. https://doi.org/10.1007/s10534-020-00236-2
Rossi P, Giansanti F, Boffi A, Ajello M, Valenti P, Chiancone E, Antonini G (2002) Ca2+ binding to bovine lactoferrin enhances protein stability and influences the release of bacterial lipopolysaccharide. Biochem Cell Biol 80:41–48. https://doi.org/10.1139/o01-209
Scholes D, Hooton TM, Roberts PL, Gupta K, Stapleton AE, Stamm WE (2005) Risk factors associated with acute pyelonephritis in healthy women. Ann Intern Med 142:20–27. https://doi.org/10.7326/0003-4819-142-1-200501040-00008
Sen A (2008) Recurrent cystitis in non-pregnant women. BMJ Clin Evid 07:801
Sessa R, Di Pietro M, Filardo S, Bressan A, Rosa L, Cutone A, Frioni A, Berlutti F, Paesano R, Valenti P (2017) Effect of bovine lactoferrin on Chlamydia trachomatis infection and inflammation. Biochem Cell Biol 95(1):34–40. https://doi.org/10.1139/bcb-2016-0049
Singh PK, Parsek MR, Greenberg EP, Welsh MJ (2002) A component of innate immunity prevents bacterial biofilm development. Nature 417(6888):552–555. https://doi.org/10.1038/417552a
Soto SM, Smithson A, Martinez JA, Horcajada JP, Mensa J, Vila J (2007) Biofilm formation in uropathogenic Escherichia coli strains: relationship with prostatitis, urovirulence factors and anti-microbial resistance. J Urol 177:365–368. https://doi.org/10.1016/j.juro.2006.08.081
Subashchandrabose S, Mobley HL (2015) Back to the metal age: battle for metals at the host-pathogen interface during urinary tract infection. Metallomics 7(6):935–942. https://doi.org/10.1039/c4mt00329b
Suzuki YA, Lopez V, Lönnerdal B (2005) Mammalian lactoferrin receptors: Structure and function. Cell Mol Life Sci 62:2560–2575. https://doi.org/10.1007/s00018-005-5371-1
Suzuki YA, Wong H, Ashida KY, Schryvers AB, Loönnerdal B (2008) The N1 domain of human lactoferrin is required for internalization by caco-2 cells and targeting to the nucleus. Biochemistry 47:10915–10920. https://doi.org/10.1021/bi8012164
U.S. FDA (2014) GRN 000465 cow’s milk-derived lactoferrin; Morinaga Milk Industry Co., Ltd.: Tokyo, Japan; U.S. Food and Drug Administration (U.S. FDA), Center for Food Safety & Applied Nutrition (CFSAN), Office of Food Additive Safety: Silver Spring, MD, USA. http://www.accessdata.fda.gov/scripts/fdcc/index.cfm?set=GRASNotices&id=465. Accessed 06 Apr 2022.
Valenti P, Antonini G (2005) Lactoferrin: an important host defence against microbial and viral attack. Cell Mol Life Sci 62:2576–2587. https://doi.org/10.1007/s00018-005-5372-0
Valenti P, Frioni A, Rossi A, Ranucci S, De Fino I, Cutone A, Rosa L, Bragonzi A, Berlutti F (2017) Aerosolized bovine lactoferrin reduces neutrophils and pro-inflammatory cytokines in mouse models of Pseudomonas aeruginosa lung infections. Biochem Cell Biol 95(1):41–47. https://doi.org/10.1139/bcb-2016-0050
Vega-Bautista A, de la Garza M, Carrero JC, Campos-Rodríguez R, Godínez-Victoria M, Drago-Serrano ME (2019) The impact of lactoferrin on the growth of intestinal inhabitant bacteria. Int J Mol Sci 20:4707. https://doi.org/10.3390/ijms20194707
WHO Regional Office for Europe/European Centre for Disease Prevention and Control (2022) Antimicrobial resistance surveillance in Europe 2022–2020 data. Copenhagen: WHO Regional Office for Europe. https://www.ecdc.europa.eu/en/publications-data/antimicrobial-resistance-surveillance-europe-2022-2020-data. Accessed 06 Apr 2022
Acknowledgements
This research received no external funding.
Author information
Authors and Affiliations
Contributions
Conceptualization AMC, RP, PV and LR; Formal analysis, GT; Investigation, ALC, RP, SS, FV, EN, GP, VC, MM, PF, PV, and LR; Writing: original draft, AMC, GT, PV, and LR; Writing-review & editing: CL, MPC, AMC, PV and LR. All authors read and approved the final version.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Conte, A.L., Longhi, C., Conte, M.P. et al. Effect of bovine lactoferrin on recurrent urinary tract infections: in vitro and in vivo evidences. Biometals 36, 491–507 (2023). https://doi.org/10.1007/s10534-022-00409-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10534-022-00409-1