Omp31 plays an important role on outer membrane properties and intracellular survival of Brucella melitensis in murine macrophages and HeLa cells
| Autor: | L Verdiguel-Fernández, A Castañeda-Ramírez, Francisco Javier Basurto-Alcantara, R Oropeza-Navarro, Antonio Verdugo-Rodríguez |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Virulence Factors 030106 microbiology Mutant Virulence Brucella Microbial Sensitivity Tests Biochemistry Microbiology Brucellosis 03 medical and health sciences Mice Cell Line Tumor Genetics medicine Brucella melitensis Animals Humans Molecular Biology Polymyxin B biology Intracellular parasite Macrophages General Medicine biology.organism_classification 030104 developmental biology Mutation Bacterial outer membrane Intracellular medicine.drug Bacterial Outer Membrane Proteins Deoxycholic Acid HeLa Cells Plasmids |
| Zdroj: | Archives of microbiology. 199(7) |
| ISSN: | 1432-072X |
| Popis: | Brucellosis is an infectious disease that affects practically all species of mammals, including human, and is a major zoonosis worldwide. Brucella spp. are facultative intracellular pathogens that have the ability to survive and multiply in phagocytic and nonphagocytic cells such as trophoblast and epithelial cells. Among the six recognized species of the genus Brucella, Brucella melitensis is the main etiological agent involved in goat brucellosis and is also the most pathogenic for human. It causes significant losses in livestock production as a result of abortions, metritis, infertility, and birth of weak animals. Outer membrane proteins (OMPs) are exposed on the bacterial surface and are in contact with cells and effectors of the host immune response, whereby they could be important virulence factors of Brucella species. To evaluate this hypothesis, the gene encoding for the major outer membrane protein Omp31 was amplified, cloned into pUC18 plasmid, and inactivated by inserting a kanamycin cassette, rendering pLVM31 plasmid which was transformed into B. melitensis wild-type strain to obtain LVM31 mutant strain. The Outer membrane (OM) properties of the mutant strain were compared with B. melitensis Bm133 wild-type and B. melitensis Rev1 vaccine strains, in assessing its susceptibility to polymyxin B, sodium deoxycholate, and nonimmune serum. The mutant strain was assessed in vitro with survival assays in murine macrophages J774.A1 and HeLa cells. Our results demonstrate that LVM31 mutant is more susceptible to polymyxin B, sodium deoxycholate, and nonimmune serum than control strains; moreover, Omp31 mutation caused a decrease in the internalization and a significant decrease in the intracellular survival compared with the reference strains in both cell lines. These results allow us to conclude that Omp31 is important for maintaining OM integrity, but also it is necessary for bacterial internalization, establishment and development of an optimal replication niche, and essential for survival and intracellular multiplication. |
| Databáze: | OpenAIRE |
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