A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens
ABSTRACTBorrelia burgdorferi, the pathogen of Lyme disease, differentially produces many outer surface proteins (Osp), some of which represent the most abundant membrane proteins, such as OspA, OspB, and OspC. In cultured bacteria, these proteins can account for a substantial fraction of the total c...
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2024-01-01
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| Series: | mSystems |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/msystems.01087-23 |
| _version_ | 1827376005670502400 |
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| author | Ozlem Buyuktanir Yaş Adam S. Coleman Rachel M. Lipman Kavita Sharma Sajith Raghunandanan Fuad Alanazi Vipin S. Rana Chrysoula Kitsou Xiuli Yang Utpal Pal |
| author_facet | Ozlem Buyuktanir Yaş Adam S. Coleman Rachel M. Lipman Kavita Sharma Sajith Raghunandanan Fuad Alanazi Vipin S. Rana Chrysoula Kitsou Xiuli Yang Utpal Pal |
| author_sort | Ozlem Buyuktanir Yaş |
| collection | DOAJ |
| description | ABSTRACTBorrelia burgdorferi, the pathogen of Lyme disease, differentially produces many outer surface proteins (Osp), some of which represent the most abundant membrane proteins, such as OspA, OspB, and OspC. In cultured bacteria, these proteins can account for a substantial fraction of the total cellular or membrane proteins, posing challenges to the identification and analysis of non-abundant proteins, which could serve as novel pathogen detection markers or as vaccine candidates. Herein, we introduced serial mutations to remove these abundant Osps and generated a B. burgdorferi mutant deficient in OspA, OspB, and OspC in an infectious 297-isolate background, designated as OspABC− mutant. Compared to parental isolate, the mutant did not reflect growth defects in the cultured medium but showed differential mRNA expression of representative tested genes, in addition to gross changes in cellular and membrane protein profiles. The analysis of differentially detectable protein contents of the OspABC− mutant, as compared to the wild type, by two-dimensional gel electrophoresis followed by liquid chromatography-mass spectrometry, identified several spirochete proteins that are dominated by proteins of unknown functions, as well as membrane transporters, chaperons, and metabolic enzymes. We produced recombinant forms of two of these represented proteins, BBA34 and BB0238, and showed that these proteins are detectable during spirochete infection in the tick-borne murine model of Lyme borreliosis and thus serve as potential antigenic markers of the infection.IMPORTANCEThe present manuscript employed a systemic approach to identify non-abundant proteins in cultured Borrelia burgdorferi that are otherwise masked or hidden due to the overwhelming presence of abundant Osps like OspA, OspB, and OspC. As these Osps are either absent or transiently expressed in mammals, we performed a proof-of-concept study in which their removal allowed the analysis of otherwise less abundant antigens in OspABC-deficient mutants and identified several immunogenic proteins, including BBA34 and BB0238. These antigens could serve as novel vaccine candidates and/or genetic markers of Lyme borreliosis, promoting new research in the clinical diagnosis and prevention of Lyme disease. |
| first_indexed | 2024-03-08T12:02:55Z |
| format | Article |
| id | doaj.art-b22d85d751014897819968b675054f25 |
| institution | Directory Open Access Journal |
| issn | 2379-5077 |
| language | English |
| last_indexed | 2024-03-08T12:02:55Z |
| publishDate | 2024-01-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSystems |
| spelling | doaj.art-b22d85d751014897819968b675054f252024-01-23T14:00:49ZengAmerican Society for MicrobiologymSystems2379-50772024-01-019110.1128/msystems.01087-23A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogensOzlem Buyuktanir Yaş0Adam S. Coleman1Rachel M. Lipman2Kavita Sharma3Sajith Raghunandanan4Fuad Alanazi5Vipin S. Rana6Chrysoula Kitsou7Xiuli Yang8Utpal Pal9Department of Microbiology, Faculty of Veterinary Medicine, Ondokuz Mayis University, Samsun, TurkeyDepartment of Veterinary Medicine, University of Maryland, College Park, Maryland, USADepartment of Veterinary Medicine, University of Maryland, College Park, Maryland, USADepartment of Veterinary Medicine, University of Maryland, College Park, Maryland, USADepartment of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Veterinary Medicine, University of Maryland, College Park, Maryland, USADepartment of Veterinary Medicine, University of Maryland, College Park, Maryland, USADepartment of Veterinary Medicine, University of Maryland, College Park, Maryland, USADepartment of Veterinary Medicine, University of Maryland, College Park, Maryland, USAABSTRACTBorrelia burgdorferi, the pathogen of Lyme disease, differentially produces many outer surface proteins (Osp), some of which represent the most abundant membrane proteins, such as OspA, OspB, and OspC. In cultured bacteria, these proteins can account for a substantial fraction of the total cellular or membrane proteins, posing challenges to the identification and analysis of non-abundant proteins, which could serve as novel pathogen detection markers or as vaccine candidates. Herein, we introduced serial mutations to remove these abundant Osps and generated a B. burgdorferi mutant deficient in OspA, OspB, and OspC in an infectious 297-isolate background, designated as OspABC− mutant. Compared to parental isolate, the mutant did not reflect growth defects in the cultured medium but showed differential mRNA expression of representative tested genes, in addition to gross changes in cellular and membrane protein profiles. The analysis of differentially detectable protein contents of the OspABC− mutant, as compared to the wild type, by two-dimensional gel electrophoresis followed by liquid chromatography-mass spectrometry, identified several spirochete proteins that are dominated by proteins of unknown functions, as well as membrane transporters, chaperons, and metabolic enzymes. We produced recombinant forms of two of these represented proteins, BBA34 and BB0238, and showed that these proteins are detectable during spirochete infection in the tick-borne murine model of Lyme borreliosis and thus serve as potential antigenic markers of the infection.IMPORTANCEThe present manuscript employed a systemic approach to identify non-abundant proteins in cultured Borrelia burgdorferi that are otherwise masked or hidden due to the overwhelming presence of abundant Osps like OspA, OspB, and OspC. As these Osps are either absent or transiently expressed in mammals, we performed a proof-of-concept study in which their removal allowed the analysis of otherwise less abundant antigens in OspABC-deficient mutants and identified several immunogenic proteins, including BBA34 and BB0238. These antigens could serve as novel vaccine candidates and/or genetic markers of Lyme borreliosis, promoting new research in the clinical diagnosis and prevention of Lyme disease.https://journals.asm.org/doi/10.1128/msystems.01087-23Borrelia burgdorferiLyme diseasemutagenesis |
| spellingShingle | Ozlem Buyuktanir Yaş Adam S. Coleman Rachel M. Lipman Kavita Sharma Sajith Raghunandanan Fuad Alanazi Vipin S. Rana Chrysoula Kitsou Xiuli Yang Utpal Pal A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens mSystems Borrelia burgdorferi Lyme disease mutagenesis |
| title | A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens |
| title_full | A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens |
| title_fullStr | A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens |
| title_full_unstemmed | A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens |
| title_short | A systemic approach to identify non-abundant immunogenic proteins in Lyme disease pathogens |
| title_sort | systemic approach to identify non abundant immunogenic proteins in lyme disease pathogens |
| topic | Borrelia burgdorferi Lyme disease mutagenesis |
| url | https://journals.asm.org/doi/10.1128/msystems.01087-23 |
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