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Recombinant Salmonella Expressing Burkholderia mallei LPS O Antigen Provides Protection in a Murine Model of Melioidosis and Glanders.

Moustafa DA, Scarff JM, Garcia PP, Cassidy SK, DiGiandomenico A, Waag DM, Inzana TJ, Goldberg JB - PLoS ONE (2015)

Bottom Line: Consequently efforts are directed towards the development of an efficacious and safe vaccine.Lipopolysaccharide (LPS) is an immunodominant antigen and potent stimulator of host immune responses.These results suggest that live-attenuated SL3261 expressing B. mallei O antigen is a promising platform for developing a safe and effective vaccine.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Cancer Biology, University of Virginia, Charlottesville, Virginia, United States of America; Department of Pediatrics, Emory University School of Medicine and Children's Hospital of Atlanta, Inc., Atlanta, Georgia, United States of America.

ABSTRACT
Burkholderia pseudomallei and Burkholderia mallei are the etiologic agents of melioidosis and glanders, respectively. These bacteria are highly infectious via the respiratory route and can cause severe and often fatal diseases in humans and animals. Both species are considered potential agents of biological warfare; they are classified as category B priority pathogens. Currently there are no human or veterinary vaccines available against these pathogens. Consequently efforts are directed towards the development of an efficacious and safe vaccine. Lipopolysaccharide (LPS) is an immunodominant antigen and potent stimulator of host immune responses. B. mallei express LPS that is structurally similar to that expressed by B. pseudomallei, suggesting the possibility of constructing a single protective vaccine against melioidosis and glanders. Previous studies of others have shown that antibodies against B. mallei or B. pseudomallei LPS partially protect mice against subsequent lethal virulent Burkholderia challenge. In this study, we evaluated the protective efficacy of recombinant Salmonella enterica serovar Typhimurium SL3261 expressing B. mallei O antigen against lethal intranasal infection with Burkholderia thailandensis, a surrogate for biothreat Burkholderia spp. in a murine model that mimics melioidosis and glanders. All vaccine-immunized mice developed a specific antibody response to B. mallei and B. pseudomallei O antigen and to B. thailandensis and were significantly protected against challenge with a lethal dose of B. thailandensis. These results suggest that live-attenuated SL3261 expressing B. mallei O antigen is a promising platform for developing a safe and effective vaccine.

No MeSH data available.


Related in: MedlinePlus

Analysis of LPS expression and immunoreactivity.(A) LPS extracted from B. mallei (Lane 2), SL3261 (Lane 3), and SL3261/p1C3 (Lane 4) was subjected to SDS-PAGE followed by immunoblotting analysis using 5C8-1C3 anti-B. mallei-specific LPS monoclonal antibody (mAb). Molecular weight marker is shown in Lane 1. (B) LPS extracted from B. mallei (B.m.) and B. pseudomallei (B.p) was run, subjected to SDS-PAGE followed by immunoblot using sera collected from SL3261 (vector)-immunized mice; 3B3-5, anti-B. pseudomallei mAb; sera collected from SL3261/p1C3 (vaccine)-immunized mice; and 5C8-1C3, anti-B. mallei mAb. Molecular weight ladder (L).
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pone.0132032.g001: Analysis of LPS expression and immunoreactivity.(A) LPS extracted from B. mallei (Lane 2), SL3261 (Lane 3), and SL3261/p1C3 (Lane 4) was subjected to SDS-PAGE followed by immunoblotting analysis using 5C8-1C3 anti-B. mallei-specific LPS monoclonal antibody (mAb). Molecular weight marker is shown in Lane 1. (B) LPS extracted from B. mallei (B.m.) and B. pseudomallei (B.p) was run, subjected to SDS-PAGE followed by immunoblot using sera collected from SL3261 (vector)-immunized mice; 3B3-5, anti-B. pseudomallei mAb; sera collected from SL3261/p1C3 (vaccine)-immunized mice; and 5C8-1C3, anti-B. mallei mAb. Molecular weight ladder (L).

Mentions: Plasmid p1C3, which contains the genes that encode proteins for production of the B. mallei O antigen, was transferred to the live-attenuated vaccine strain Salmonella enterica serovar Typhimurium SL3261. In order to determine whether SL3261 was capable of expressing the B. mallei O antigen, LPS was extracted from strains SL3261 and SL3261/p1C3 and analyzed by immunoblotting with a monoclonal antibody (mAb) specific for the B. mallei O antigen (Fig 1A). Our results indicate that SL3261/pIC3 is capable of expressing the B. mallei O-antigen (Fig 1A).


Recombinant Salmonella Expressing Burkholderia mallei LPS O Antigen Provides Protection in a Murine Model of Melioidosis and Glanders.

Moustafa DA, Scarff JM, Garcia PP, Cassidy SK, DiGiandomenico A, Waag DM, Inzana TJ, Goldberg JB - PLoS ONE (2015)

Analysis of LPS expression and immunoreactivity.(A) LPS extracted from B. mallei (Lane 2), SL3261 (Lane 3), and SL3261/p1C3 (Lane 4) was subjected to SDS-PAGE followed by immunoblotting analysis using 5C8-1C3 anti-B. mallei-specific LPS monoclonal antibody (mAb). Molecular weight marker is shown in Lane 1. (B) LPS extracted from B. mallei (B.m.) and B. pseudomallei (B.p) was run, subjected to SDS-PAGE followed by immunoblot using sera collected from SL3261 (vector)-immunized mice; 3B3-5, anti-B. pseudomallei mAb; sera collected from SL3261/p1C3 (vaccine)-immunized mice; and 5C8-1C3, anti-B. mallei mAb. Molecular weight ladder (L).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4492786&req=5

pone.0132032.g001: Analysis of LPS expression and immunoreactivity.(A) LPS extracted from B. mallei (Lane 2), SL3261 (Lane 3), and SL3261/p1C3 (Lane 4) was subjected to SDS-PAGE followed by immunoblotting analysis using 5C8-1C3 anti-B. mallei-specific LPS monoclonal antibody (mAb). Molecular weight marker is shown in Lane 1. (B) LPS extracted from B. mallei (B.m.) and B. pseudomallei (B.p) was run, subjected to SDS-PAGE followed by immunoblot using sera collected from SL3261 (vector)-immunized mice; 3B3-5, anti-B. pseudomallei mAb; sera collected from SL3261/p1C3 (vaccine)-immunized mice; and 5C8-1C3, anti-B. mallei mAb. Molecular weight ladder (L).
Mentions: Plasmid p1C3, which contains the genes that encode proteins for production of the B. mallei O antigen, was transferred to the live-attenuated vaccine strain Salmonella enterica serovar Typhimurium SL3261. In order to determine whether SL3261 was capable of expressing the B. mallei O antigen, LPS was extracted from strains SL3261 and SL3261/p1C3 and analyzed by immunoblotting with a monoclonal antibody (mAb) specific for the B. mallei O antigen (Fig 1A). Our results indicate that SL3261/pIC3 is capable of expressing the B. mallei O-antigen (Fig 1A).

Bottom Line: Consequently efforts are directed towards the development of an efficacious and safe vaccine.Lipopolysaccharide (LPS) is an immunodominant antigen and potent stimulator of host immune responses.These results suggest that live-attenuated SL3261 expressing B. mallei O antigen is a promising platform for developing a safe and effective vaccine.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Cancer Biology, University of Virginia, Charlottesville, Virginia, United States of America; Department of Pediatrics, Emory University School of Medicine and Children's Hospital of Atlanta, Inc., Atlanta, Georgia, United States of America.

ABSTRACT
Burkholderia pseudomallei and Burkholderia mallei are the etiologic agents of melioidosis and glanders, respectively. These bacteria are highly infectious via the respiratory route and can cause severe and often fatal diseases in humans and animals. Both species are considered potential agents of biological warfare; they are classified as category B priority pathogens. Currently there are no human or veterinary vaccines available against these pathogens. Consequently efforts are directed towards the development of an efficacious and safe vaccine. Lipopolysaccharide (LPS) is an immunodominant antigen and potent stimulator of host immune responses. B. mallei express LPS that is structurally similar to that expressed by B. pseudomallei, suggesting the possibility of constructing a single protective vaccine against melioidosis and glanders. Previous studies of others have shown that antibodies against B. mallei or B. pseudomallei LPS partially protect mice against subsequent lethal virulent Burkholderia challenge. In this study, we evaluated the protective efficacy of recombinant Salmonella enterica serovar Typhimurium SL3261 expressing B. mallei O antigen against lethal intranasal infection with Burkholderia thailandensis, a surrogate for biothreat Burkholderia spp. in a murine model that mimics melioidosis and glanders. All vaccine-immunized mice developed a specific antibody response to B. mallei and B. pseudomallei O antigen and to B. thailandensis and were significantly protected against challenge with a lethal dose of B. thailandensis. These results suggest that live-attenuated SL3261 expressing B. mallei O antigen is a promising platform for developing a safe and effective vaccine.

No MeSH data available.


Related in: MedlinePlus