The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infections

Lactobacillus crispatus is renowned for its antimicrobial properties, and some strains are used to treat vaginal dysbiosis, although the mechanisms underlying the antimicrobial properties remain elusive. We isolated L. crispatus M247 (LcM247) from a commercially available probiotic product Crispact®...

Full description

Saved in:
Bibliographic Details
Main Authors: Giulia Santarelli, Roberto Rosato, Michela Cicchinelli, Federica Iavarone, Andrea Urbani, Maurizio Sanguinetti, Giovanni Delogu, Flavio De Maio
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Cellular and Infection Microbiology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fcimb.2025.1586442/full
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849331211712856064
author Giulia Santarelli
Roberto Rosato
Michela Cicchinelli
Federica Iavarone
Federica Iavarone
Andrea Urbani
Andrea Urbani
Maurizio Sanguinetti
Maurizio Sanguinetti
Giovanni Delogu
Giovanni Delogu
Flavio De Maio
author_facet Giulia Santarelli
Roberto Rosato
Michela Cicchinelli
Federica Iavarone
Federica Iavarone
Andrea Urbani
Andrea Urbani
Maurizio Sanguinetti
Maurizio Sanguinetti
Giovanni Delogu
Giovanni Delogu
Flavio De Maio
author_sort Giulia Santarelli
collection DOAJ
description Lactobacillus crispatus is renowned for its antimicrobial properties, and some strains are used to treat vaginal dysbiosis, although the mechanisms underlying the antimicrobial properties remain elusive. We isolated L. crispatus M247 (LcM247) from a commercially available probiotic product Crispact® and tested its antimicrobial activity against selected pathobionts such as Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus agalactiae, Enterococcus faecalis and Candida albicans using both cocultures and testing the antimicrobial activity of cell-free supernatant (CFS) obtained from the culture of the probiotic strain. Furthermore, we demonstrate that CFS antimicrobial activity is pH dependent and that it is not affected by temperature and proteinase K treatment. Proteomic analysis suggests that this activity is mediated by S-layer secreted proteins. In a series of in vitro infection models, we infected Henrietta Lacks’ cervical eukaryotic cancer cells (HeLa) with E. coli, S. agalactiae and C. albicans at specific multiplicities of infection (MOIs) before the administration of LcM247, CFS, gentamicin or fluconazole alone or in combination with LcM247/CFS. We observed a slight decrease in the microbial burden following LcM247 administration, while treatment with CFS significantly reduced microbial growth compared to control and antimicrobial compounds. These results highlight the antimicrobial properties of LcM247 and its CFS and the likely mechanism of action that contributes to the eradication of common pathobionts. We show that actively replicating LcM247 is less efficient than its CFS, so the oral administration of LcM247 may result in treatment failure. Finally, the use of CFS may result in an upswing of the host Lactobacillus strains and promote the engraftment of Lactobacillus probiotic treatments.
format Article
id doaj-art-253be421ecf545b39f813fbcbb627bfa
institution Kabale University
issn 2235-2988
language English
publishDate 2025-06-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Cellular and Infection Microbiology
spelling doaj-art-253be421ecf545b39f813fbcbb627bfa2025-08-20T03:46:42ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-06-011510.3389/fcimb.2025.15864421586442The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infectionsGiulia Santarelli0Roberto Rosato1Michela Cicchinelli2Federica Iavarone3Federica Iavarone4Andrea Urbani5Andrea Urbani6Maurizio Sanguinetti7Maurizio Sanguinetti8Giovanni Delogu9Giovanni Delogu10Flavio De Maio11Dipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, Rome, ItalyDipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, Rome, ItalyDipartimento di Scienze di Laboratorio ed Ematologiche, Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, ItalyDipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, Rome, ItalyDipartimento di Scienze di Laboratorio ed Ematologiche, Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, ItalyDipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, Rome, ItalyDipartimento di Scienze di Laboratorio ed Ematologiche, Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, ItalyDipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, Rome, ItalyDipartimento di Scienze di Laboratorio ed Ematologiche, Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, ItalyDipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, Rome, ItalyMater Olbia Hospital, Olbia, ItalyDipartimento di Scienze di Laboratorio ed Ematologiche, Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome, ItalyLactobacillus crispatus is renowned for its antimicrobial properties, and some strains are used to treat vaginal dysbiosis, although the mechanisms underlying the antimicrobial properties remain elusive. We isolated L. crispatus M247 (LcM247) from a commercially available probiotic product Crispact® and tested its antimicrobial activity against selected pathobionts such as Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus agalactiae, Enterococcus faecalis and Candida albicans using both cocultures and testing the antimicrobial activity of cell-free supernatant (CFS) obtained from the culture of the probiotic strain. Furthermore, we demonstrate that CFS antimicrobial activity is pH dependent and that it is not affected by temperature and proteinase K treatment. Proteomic analysis suggests that this activity is mediated by S-layer secreted proteins. In a series of in vitro infection models, we infected Henrietta Lacks’ cervical eukaryotic cancer cells (HeLa) with E. coli, S. agalactiae and C. albicans at specific multiplicities of infection (MOIs) before the administration of LcM247, CFS, gentamicin or fluconazole alone or in combination with LcM247/CFS. We observed a slight decrease in the microbial burden following LcM247 administration, while treatment with CFS significantly reduced microbial growth compared to control and antimicrobial compounds. These results highlight the antimicrobial properties of LcM247 and its CFS and the likely mechanism of action that contributes to the eradication of common pathobionts. We show that actively replicating LcM247 is less efficient than its CFS, so the oral administration of LcM247 may result in treatment failure. Finally, the use of CFS may result in an upswing of the host Lactobacillus strains and promote the engraftment of Lactobacillus probiotic treatments.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1586442/fullLactobacillus crispatusvaginal infectionsprobioticscell-free supernatantLcM247
spellingShingle Giulia Santarelli
Roberto Rosato
Michela Cicchinelli
Federica Iavarone
Federica Iavarone
Andrea Urbani
Andrea Urbani
Maurizio Sanguinetti
Maurizio Sanguinetti
Giovanni Delogu
Giovanni Delogu
Flavio De Maio
The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infections
Frontiers in Cellular and Infection Microbiology
Lactobacillus crispatus
vaginal infections
probiotics
cell-free supernatant
LcM247
title The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infections
title_full The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infections
title_fullStr The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infections
title_full_unstemmed The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infections
title_short The activity of cell-free supernatant of Lactobacillus crispatus M247: a promising treatment against vaginal infections
title_sort activity of cell free supernatant of lactobacillus crispatus m247 a promising treatment against vaginal infections
topic Lactobacillus crispatus
vaginal infections
probiotics
cell-free supernatant
LcM247
url https://www.frontiersin.org/articles/10.3389/fcimb.2025.1586442/full
work_keys_str_mv AT giuliasantarelli theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT robertorosato theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT michelacicchinelli theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT federicaiavarone theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT federicaiavarone theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT andreaurbani theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT andreaurbani theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT mauriziosanguinetti theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT mauriziosanguinetti theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT giovannidelogu theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT giovannidelogu theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT flaviodemaio theactivityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT giuliasantarelli activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT robertorosato activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT michelacicchinelli activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT federicaiavarone activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT federicaiavarone activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT andreaurbani activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT andreaurbani activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT mauriziosanguinetti activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT mauriziosanguinetti activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT giovannidelogu activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT giovannidelogu activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections
AT flaviodemaio activityofcellfreesupernatantoflactobacilluscrispatusm247apromisingtreatmentagainstvaginalinfections