Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach.
A unique hallmark of tuberculosis is the granulomatous lesions formed in the lung. Granulomas can be heterogeneous in nature and can develop a necrotic, hypoxic core which is surrounded by an acellular, fibrotic rim. Studying bacilli in this in vivo microenvironment is problematic as Mycobacterium t...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2010-06-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2885421?pdf=render |
| _version_ | 1818416138125574144 |
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| author | Gavin J Ryan Donald R Hoff Emily R Driver Martin I Voskuil Mercedes Gonzalez-Juarrero Randall J Basaraba Dean C Crick John S Spencer Anne J Lenaerts |
| author_facet | Gavin J Ryan Donald R Hoff Emily R Driver Martin I Voskuil Mercedes Gonzalez-Juarrero Randall J Basaraba Dean C Crick John S Spencer Anne J Lenaerts |
| author_sort | Gavin J Ryan |
| collection | DOAJ |
| description | A unique hallmark of tuberculosis is the granulomatous lesions formed in the lung. Granulomas can be heterogeneous in nature and can develop a necrotic, hypoxic core which is surrounded by an acellular, fibrotic rim. Studying bacilli in this in vivo microenvironment is problematic as Mycobacterium tuberculosis can change its phenotype and also become acid-fast negative. Under in vitro models of differing environments, M. tuberculosis alters its metabolism, transcriptional profile and rate of replication. In this study, we investigated whether these phenotypic adaptations of M. tuberculosis are unique for certain environmental conditions and if they could therefore be used as differential markers. Bacilli were studied using fluorescent acid-fast auramine-rhodamine targeting the mycolic acid containing cell wall, and immunofluorescence targeting bacterial proteins using an anti-M. tuberculosis whole cell lysate polyclonal antibody. These techniques were combined and simultaneously applied to M. tuberculosis in vitro culture samples and to lung sections of M. tuberculosis infected mice and guinea pigs. Two phenotypically different subpopulations of M. tuberculosis were found in stationary culture whilst three subpopulations were found in hypoxic culture and in lung sections. Bacilli were either exclusively acid-fast positive, exclusively immunofluorescent positive or acid-fast and immunofluorescent positive. These results suggest that M. tuberculosis exists as multiple populations in most conditions, even within seemingly a single microenvironment. This is relevant information for approaches that study bacillary characteristics in pooled samples (using lipidomics and proteomics) as well as in M. tuberculosis drug development. |
| first_indexed | 2024-12-14T11:46:07Z |
| format | Article |
| id | doaj.art-442a27b408104ffd819da86b3a48cd29 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T11:46:07Z |
| publishDate | 2010-06-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-442a27b408104ffd819da86b3a48cd292022-12-21T23:02:34ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-06-0156e1110810.1371/journal.pone.0011108Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach.Gavin J RyanDonald R HoffEmily R DriverMartin I VoskuilMercedes Gonzalez-JuarreroRandall J BasarabaDean C CrickJohn S SpencerAnne J LenaertsA unique hallmark of tuberculosis is the granulomatous lesions formed in the lung. Granulomas can be heterogeneous in nature and can develop a necrotic, hypoxic core which is surrounded by an acellular, fibrotic rim. Studying bacilli in this in vivo microenvironment is problematic as Mycobacterium tuberculosis can change its phenotype and also become acid-fast negative. Under in vitro models of differing environments, M. tuberculosis alters its metabolism, transcriptional profile and rate of replication. In this study, we investigated whether these phenotypic adaptations of M. tuberculosis are unique for certain environmental conditions and if they could therefore be used as differential markers. Bacilli were studied using fluorescent acid-fast auramine-rhodamine targeting the mycolic acid containing cell wall, and immunofluorescence targeting bacterial proteins using an anti-M. tuberculosis whole cell lysate polyclonal antibody. These techniques were combined and simultaneously applied to M. tuberculosis in vitro culture samples and to lung sections of M. tuberculosis infected mice and guinea pigs. Two phenotypically different subpopulations of M. tuberculosis were found in stationary culture whilst three subpopulations were found in hypoxic culture and in lung sections. Bacilli were either exclusively acid-fast positive, exclusively immunofluorescent positive or acid-fast and immunofluorescent positive. These results suggest that M. tuberculosis exists as multiple populations in most conditions, even within seemingly a single microenvironment. This is relevant information for approaches that study bacillary characteristics in pooled samples (using lipidomics and proteomics) as well as in M. tuberculosis drug development.http://europepmc.org/articles/PMC2885421?pdf=render |
| spellingShingle | Gavin J Ryan Donald R Hoff Emily R Driver Martin I Voskuil Mercedes Gonzalez-Juarrero Randall J Basaraba Dean C Crick John S Spencer Anne J Lenaerts Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach. PLoS ONE |
| title | Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach. |
| title_full | Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach. |
| title_fullStr | Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach. |
| title_full_unstemmed | Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach. |
| title_short | Multiple M. tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual-staining approach. |
| title_sort | multiple m tuberculosis phenotypes in mouse and guinea pig lung tissue revealed by a dual staining approach |
| url | http://europepmc.org/articles/PMC2885421?pdf=render |
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