Cargando…

Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37

The antimicrobial peptide LL-37 inhibits the growth of the major human pathogen Mycobacterium tuberculosis (Mtb), but the mechanism of the peptide–pathogen interaction inside human macrophages remains unclear. Super-resolution imaging techniques provide a novel opportunity to visualize these interac...

Descripción completa

Detalles Bibliográficos
Autores principales: Deshpande, Dhruva, Grieshober, Mark, Wondany, Fanny, Gerbl, Fabian, Noschka, Reiner, Michaelis, Jens, Stenger, Steffen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555347/
https://www.ncbi.nlm.nih.gov/pubmed/32937921
http://dx.doi.org/10.3390/ijms21186741
_version_ 1783593986712141824
author Deshpande, Dhruva
Grieshober, Mark
Wondany, Fanny
Gerbl, Fabian
Noschka, Reiner
Michaelis, Jens
Stenger, Steffen
author_facet Deshpande, Dhruva
Grieshober, Mark
Wondany, Fanny
Gerbl, Fabian
Noschka, Reiner
Michaelis, Jens
Stenger, Steffen
author_sort Deshpande, Dhruva
collection PubMed
description The antimicrobial peptide LL-37 inhibits the growth of the major human pathogen Mycobacterium tuberculosis (Mtb), but the mechanism of the peptide–pathogen interaction inside human macrophages remains unclear. Super-resolution imaging techniques provide a novel opportunity to visualize these interactions on a molecular level. Here, we adapt the super-resolution technique of stimulated emission depletion (STED) microscopy to study the uptake, intracellular localization and interaction of LL-37 with macrophages and virulent Mtb. We demonstrate that LL-37 is internalized by both uninfected and Mtb infected primary human macrophages. The peptide localizes in the membrane of early endosomes and lysosomes, the compartment in which mycobacteria reside. Functionally, LL-37 disrupts the cell wall of intra- and extracellular Mtb, resulting in the killing of the pathogen. In conclusion, we introduce STED microscopy as an innovative and informative tool for studying host–pathogen–peptide interactions, clearly extending the possibilities of conventional confocal microscopy.
format Online
Article
Text
id pubmed-7555347
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-75553472020-10-19 Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37 Deshpande, Dhruva Grieshober, Mark Wondany, Fanny Gerbl, Fabian Noschka, Reiner Michaelis, Jens Stenger, Steffen Int J Mol Sci Article The antimicrobial peptide LL-37 inhibits the growth of the major human pathogen Mycobacterium tuberculosis (Mtb), but the mechanism of the peptide–pathogen interaction inside human macrophages remains unclear. Super-resolution imaging techniques provide a novel opportunity to visualize these interactions on a molecular level. Here, we adapt the super-resolution technique of stimulated emission depletion (STED) microscopy to study the uptake, intracellular localization and interaction of LL-37 with macrophages and virulent Mtb. We demonstrate that LL-37 is internalized by both uninfected and Mtb infected primary human macrophages. The peptide localizes in the membrane of early endosomes and lysosomes, the compartment in which mycobacteria reside. Functionally, LL-37 disrupts the cell wall of intra- and extracellular Mtb, resulting in the killing of the pathogen. In conclusion, we introduce STED microscopy as an innovative and informative tool for studying host–pathogen–peptide interactions, clearly extending the possibilities of conventional confocal microscopy. MDPI 2020-09-14 /pmc/articles/PMC7555347/ /pubmed/32937921 http://dx.doi.org/10.3390/ijms21186741 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Deshpande, Dhruva
Grieshober, Mark
Wondany, Fanny
Gerbl, Fabian
Noschka, Reiner
Michaelis, Jens
Stenger, Steffen
Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37
title Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37
title_full Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37
title_fullStr Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37
title_full_unstemmed Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37
title_short Super-Resolution Microscopy Reveals a Direct Interaction of Intracellular Mycobacterium tuberculosis with the Antimicrobial Peptide LL-37
title_sort super-resolution microscopy reveals a direct interaction of intracellular mycobacterium tuberculosis with the antimicrobial peptide ll-37
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555347/
https://www.ncbi.nlm.nih.gov/pubmed/32937921
http://dx.doi.org/10.3390/ijms21186741
work_keys_str_mv AT deshpandedhruva superresolutionmicroscopyrevealsadirectinteractionofintracellularmycobacteriumtuberculosiswiththeantimicrobialpeptidell37
AT grieshobermark superresolutionmicroscopyrevealsadirectinteractionofintracellularmycobacteriumtuberculosiswiththeantimicrobialpeptidell37
AT wondanyfanny superresolutionmicroscopyrevealsadirectinteractionofintracellularmycobacteriumtuberculosiswiththeantimicrobialpeptidell37
AT gerblfabian superresolutionmicroscopyrevealsadirectinteractionofintracellularmycobacteriumtuberculosiswiththeantimicrobialpeptidell37
AT noschkareiner superresolutionmicroscopyrevealsadirectinteractionofintracellularmycobacteriumtuberculosiswiththeantimicrobialpeptidell37
AT michaelisjens superresolutionmicroscopyrevealsadirectinteractionofintracellularmycobacteriumtuberculosiswiththeantimicrobialpeptidell37
AT stengersteffen superresolutionmicroscopyrevealsadirectinteractionofintracellularmycobacteriumtuberculosiswiththeantimicrobialpeptidell37