Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach
IntroductionThe environmental bacterium Burkholderia pseudomallei causes the often fatal and massively underreported infectious disease melioidosis. Antigens inducing protective immunity in experimental models have recently been identified and serodiagnostic tools have been improved. However, furthe...
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| Format: | Article |
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Frontiers Media S.A.
2023-12-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1294113/full |
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| author | Gabriel E. Wagner Thomas Franz Paul Stanjek Dirk Albrecht Michaela Lipp Susanna J. Dunachie Susanna J. Dunachie Susanna J. Dunachie Esther Föderl-Höbenreich Esther Föderl-Höbenreich Katharina Riedel Anne Kohler Ivo Steinmetz Ivo Steinmetz Christian Kohler |
| author_facet | Gabriel E. Wagner Thomas Franz Paul Stanjek Dirk Albrecht Michaela Lipp Susanna J. Dunachie Susanna J. Dunachie Susanna J. Dunachie Esther Föderl-Höbenreich Esther Föderl-Höbenreich Katharina Riedel Anne Kohler Ivo Steinmetz Ivo Steinmetz Christian Kohler |
| author_sort | Gabriel E. Wagner |
| collection | DOAJ |
| description | IntroductionThe environmental bacterium Burkholderia pseudomallei causes the often fatal and massively underreported infectious disease melioidosis. Antigens inducing protective immunity in experimental models have recently been identified and serodiagnostic tools have been improved. However, further elucidation of the antigenic repertoire of B. pseudomallei during human infection for diagnostic and vaccine purposes is required. The adaptation of B. pseudomallei to very different habitats is reflected by a huge genome and a selective transcriptional response to a variety of conditions. We, therefore, hypothesized that exposure of B. pseudomallei to culture conditions mimicking habitats encountered in the human host might unravel novel antigens that are recognized by melioidosis patients.Methods and resultsIn this study, B. pseudomallei was exposed to various stress and growth conditions, including anaerobiosis, acid stress, oxidative stress, iron starvation and osmotic stress. Immunogenic proteins were identified by probing two-dimensional Western blots of B. pseudomallei intracellular and extracellular protein extracts with sera from melioidosis patients and controls and subsequent MALDI-TOF MS. Among B. pseudomallei specific immunogenic signals, 90 % (55/61) of extracellular immunogenic proteins were identified by acid, osmotic or oxidative stress. A total of 84 % (44/52) of intracellular antigens originated from the stationary growth phase, acidic, oxidative and anaerobic conditions. The majority of the extracellular and intracellular protein antigens were identified in only one of the various stress conditions. Sixty-three immunoreactive proteins and an additional 38 candidates from a literature screening were heterologously expressed and subjected to dot blot analysis using melioidosis sera and controls. Our experiments confirmed melioidosis-specific signals in 58 of our immunoproteome candidates. These include 15 antigens with average signal ratios (melioidosis:controls) greater than 10 and another 26 with average ratios greater than 5, including new promising serodiagnostic candidates with a very high signal-to-noise ratio.ConclusionOur study shows that a comprehensive B. pseudomallei immunoproteomics approach, using conditions which are likely to be encountered during infection, can identify novel antibody targets previously unrecognized in human melioidosis. |
| first_indexed | 2024-03-09T01:05:27Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-09T01:05:27Z |
| publishDate | 2023-12-01 |
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| series | Frontiers in Immunology |
| spelling | doaj.art-26cabcceb2f4435a9802762527a53d012023-12-11T10:00:38ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-12-011410.3389/fimmu.2023.12941131294113Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approachGabriel E. Wagner0Thomas Franz Paul Stanjek1Dirk Albrecht2Michaela Lipp3Susanna J. Dunachie4Susanna J. Dunachie5Susanna J. Dunachie6Esther Föderl-Höbenreich7Esther Föderl-Höbenreich8Katharina Riedel9Anne Kohler10Ivo Steinmetz11Ivo Steinmetz12Christian Kohler13Diagnostic and Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, AustriaFriedrich Loeffler Institute of Medical Microbiology, University Medicine, Greifswald, GermanyInstitute of Microbiology, Department of Microbial Physiology and Molecular Biology, University of Greifswald, Greifswald, GermanyDiagnostic and Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, AustriaNuffield Department of Medicine (NDM) Centre for Global Health Research, Nuffield Department of Medicine, University of Oxford, Oxford, United KingdomNational Institute for Health and Care Research (NIHR) Oxford Biomedical Centre, Oxford University Hospitals National Health Service (NHS) Foundation Trust, Oxford, United KingdomMahidol-Oxford Tropical Medicine Research Unit, Mahidol University, Bangkok, ThailandDiagnostic and Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, AustriaDiagnostic & Research Institute of Pathology, Medical University Graz, Graz, AustriaInstitute of Microbiology, Department of Microbial Physiology and Molecular Biology, University of Greifswald, Greifswald, GermanyFriedrich Loeffler Institute of Medical Microbiology, University Medicine, Greifswald, GermanyDiagnostic and Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, AustriaFriedrich Loeffler Institute of Medical Microbiology, University Medicine, Greifswald, GermanyFriedrich Loeffler Institute of Medical Microbiology, University Medicine, Greifswald, GermanyIntroductionThe environmental bacterium Burkholderia pseudomallei causes the often fatal and massively underreported infectious disease melioidosis. Antigens inducing protective immunity in experimental models have recently been identified and serodiagnostic tools have been improved. However, further elucidation of the antigenic repertoire of B. pseudomallei during human infection for diagnostic and vaccine purposes is required. The adaptation of B. pseudomallei to very different habitats is reflected by a huge genome and a selective transcriptional response to a variety of conditions. We, therefore, hypothesized that exposure of B. pseudomallei to culture conditions mimicking habitats encountered in the human host might unravel novel antigens that are recognized by melioidosis patients.Methods and resultsIn this study, B. pseudomallei was exposed to various stress and growth conditions, including anaerobiosis, acid stress, oxidative stress, iron starvation and osmotic stress. Immunogenic proteins were identified by probing two-dimensional Western blots of B. pseudomallei intracellular and extracellular protein extracts with sera from melioidosis patients and controls and subsequent MALDI-TOF MS. Among B. pseudomallei specific immunogenic signals, 90 % (55/61) of extracellular immunogenic proteins were identified by acid, osmotic or oxidative stress. A total of 84 % (44/52) of intracellular antigens originated from the stationary growth phase, acidic, oxidative and anaerobic conditions. The majority of the extracellular and intracellular protein antigens were identified in only one of the various stress conditions. Sixty-three immunoreactive proteins and an additional 38 candidates from a literature screening were heterologously expressed and subjected to dot blot analysis using melioidosis sera and controls. Our experiments confirmed melioidosis-specific signals in 58 of our immunoproteome candidates. These include 15 antigens with average signal ratios (melioidosis:controls) greater than 10 and another 26 with average ratios greater than 5, including new promising serodiagnostic candidates with a very high signal-to-noise ratio.ConclusionOur study shows that a comprehensive B. pseudomallei immunoproteomics approach, using conditions which are likely to be encountered during infection, can identify novel antibody targets previously unrecognized in human melioidosis.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1294113/fullBurkholderia pseudomalleiimmunoproteomicsmelioidosisdot blotWestern blot |
| spellingShingle | Gabriel E. Wagner Thomas Franz Paul Stanjek Dirk Albrecht Michaela Lipp Susanna J. Dunachie Susanna J. Dunachie Susanna J. Dunachie Esther Föderl-Höbenreich Esther Föderl-Höbenreich Katharina Riedel Anne Kohler Ivo Steinmetz Ivo Steinmetz Christian Kohler Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach Frontiers in Immunology Burkholderia pseudomallei immunoproteomics melioidosis dot blot Western blot |
| title | Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach |
| title_full | Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach |
| title_fullStr | Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach |
| title_full_unstemmed | Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach |
| title_short | Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach |
| title_sort | deciphering the human antibody response against burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach |
| topic | Burkholderia pseudomallei immunoproteomics melioidosis dot blot Western blot |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1294113/full |
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