Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense Mechanisms
Klebsiella pneumoniae produces capsular polysaccharides that are a crucial virulence factor protecting bacteria against innate response mechanisms of the infected host. Simultaneously, those capsules are targeted by specific bacteriophages equipped with virion-associated depolymerases able to recogn...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2018-10-01
|
| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.02517/full |
| _version_ | 1819177255815872512 |
|---|---|
| author | Grazyna Majkowska-Skrobek Agnieszka Latka Agnieszka Latka Rita Berisio Flavia Squeglia Barbara Maciejewska Yves Briers Zuzanna Drulis-Kawa |
| author_facet | Grazyna Majkowska-Skrobek Agnieszka Latka Agnieszka Latka Rita Berisio Flavia Squeglia Barbara Maciejewska Yves Briers Zuzanna Drulis-Kawa |
| author_sort | Grazyna Majkowska-Skrobek |
| collection | DOAJ |
| description | Klebsiella pneumoniae produces capsular polysaccharides that are a crucial virulence factor protecting bacteria against innate response mechanisms of the infected host. Simultaneously, those capsules are targeted by specific bacteriophages equipped with virion-associated depolymerases able to recognize and degrade these polysaccharides. We show that Klebsiella phage KP32 produces two capsule depolymerases, KP32gp37 and KP32gp38, with a high specificity for the capsular serotypes K3 and K21, respectively. Together, they determine the host spectrum of bacteriophage KP32, which is limited to strains with serotype K3 and K21. Both depolymerases form a trimeric β-structure, display moderate thermostability and function optimally under neutral to alkaline conditions. We show that both depolymerases strongly affect the virulence of K. pneumoniae with the corresponding K3 and K21 capsular serotypes. Capsule degradation renders the otherwise serum-resistant cells more prone to complement-mediated killing with up to four log reduction in serum upon exposure to KP32gp37. Decapsulated strains are also sensitized for phagocytosis with a twofold increased uptake. In addition, the intracellular survival of phagocytized cells in macrophages was significantly reduced when bacteria were previously exposed to the capsule depolymerases. Finally, depolymerase application considerably increases the lifespan of Galleria mellonella larvae infected with K. pneumoniae in a time- and strain-dependent manner. In sum, capsule depolymerases are promising antivirulence compounds that act by defeating a major resistance mechanism of K. pneumoniae against the innate immunity. |
| first_indexed | 2024-12-22T21:23:45Z |
| format | Article |
| id | doaj.art-5f592070a3fa46c2947c14db91f34e2a |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-22T21:23:45Z |
| publishDate | 2018-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-5f592070a3fa46c2947c14db91f34e2a2022-12-21T18:12:06ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-10-01910.3389/fmicb.2018.02517419180Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense MechanismsGrazyna Majkowska-Skrobek0Agnieszka Latka1Agnieszka Latka2Rita Berisio3Flavia Squeglia4Barbara Maciejewska5Yves Briers6Zuzanna Drulis-Kawa7Department of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wrocław, Wrocław, PolandDepartment of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wrocław, Wrocław, PolandLaboratory of Applied Biotechnology, Department of Biotechnology, Ghent University, Ghent, BelgiumInstitute of Biostructure and Bioimaging, Italian National Research Council, Naples, ItalyInstitute of Biostructure and Bioimaging, Italian National Research Council, Naples, ItalyDepartment of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wrocław, Wrocław, PolandLaboratory of Applied Biotechnology, Department of Biotechnology, Ghent University, Ghent, BelgiumDepartment of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wrocław, Wrocław, PolandKlebsiella pneumoniae produces capsular polysaccharides that are a crucial virulence factor protecting bacteria against innate response mechanisms of the infected host. Simultaneously, those capsules are targeted by specific bacteriophages equipped with virion-associated depolymerases able to recognize and degrade these polysaccharides. We show that Klebsiella phage KP32 produces two capsule depolymerases, KP32gp37 and KP32gp38, with a high specificity for the capsular serotypes K3 and K21, respectively. Together, they determine the host spectrum of bacteriophage KP32, which is limited to strains with serotype K3 and K21. Both depolymerases form a trimeric β-structure, display moderate thermostability and function optimally under neutral to alkaline conditions. We show that both depolymerases strongly affect the virulence of K. pneumoniae with the corresponding K3 and K21 capsular serotypes. Capsule degradation renders the otherwise serum-resistant cells more prone to complement-mediated killing with up to four log reduction in serum upon exposure to KP32gp37. Decapsulated strains are also sensitized for phagocytosis with a twofold increased uptake. In addition, the intracellular survival of phagocytized cells in macrophages was significantly reduced when bacteria were previously exposed to the capsule depolymerases. Finally, depolymerase application considerably increases the lifespan of Galleria mellonella larvae infected with K. pneumoniae in a time- and strain-dependent manner. In sum, capsule depolymerases are promising antivirulence compounds that act by defeating a major resistance mechanism of K. pneumoniae against the innate immunity.https://www.frontiersin.org/article/10.3389/fmicb.2018.02517/fullbacteriophageKlebsiella sp.capsulepolysaccharide depolymeraseinnate immunity |
| spellingShingle | Grazyna Majkowska-Skrobek Agnieszka Latka Agnieszka Latka Rita Berisio Flavia Squeglia Barbara Maciejewska Yves Briers Zuzanna Drulis-Kawa Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense Mechanisms Frontiers in Microbiology bacteriophage Klebsiella sp. capsule polysaccharide depolymerase innate immunity |
| title | Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense Mechanisms |
| title_full | Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense Mechanisms |
| title_fullStr | Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense Mechanisms |
| title_full_unstemmed | Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense Mechanisms |
| title_short | Phage-Borne Depolymerases Decrease Klebsiella pneumoniae Resistance to Innate Defense Mechanisms |
| title_sort | phage borne depolymerases decrease klebsiella pneumoniae resistance to innate defense mechanisms |
| topic | bacteriophage Klebsiella sp. capsule polysaccharide depolymerase innate immunity |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.02517/full |
| work_keys_str_mv | AT grazynamajkowskaskrobek phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms AT agnieszkalatka phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms AT agnieszkalatka phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms AT ritaberisio phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms AT flaviasqueglia phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms AT barbaramaciejewska phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms AT yvesbriers phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms AT zuzannadruliskawa phagebornedepolymerasesdecreaseklebsiellapneumoniaeresistancetoinnatedefensemechanisms |