Identification of a LolC Homologue in<i>Burkholderia pseudomallei</i>, a Novel Protective Antigen for Melioidosis

Harland, David N.; Chu, Karen K.; Haque, Ashraful; Nelson, Michelle; Walker, Nicola; Sarkar‐Tyson, Mitali; Atkins, Timothy P.; Moore, Benjamin L.; Brown, Katherine A.; Bancroft, Gregory J.; Titball, Richard W.; Atkins, Helen S.

doi:10.1128/iai.00404-07

Cited by 61 publications

(77 citation statements)

References 40 publications

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“…Considerable work has been done to identify protein targets to act as a subunit vaccine for B. pseudomallei and B. mallei (4,5,18,23,24). However, relatively few of these proteins provide suitable protection, despite being highly immunogenic.…”

Section: Discussionmentioning

confidence: 99%

Flagellar Glycosylation in Burkholderia pseudomallei and Burkholderia thailandensis

Scott¹,

Twine²,

Fulton³

et al. 2011

Journal of Bacteriology

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Glycosylation of proteins is known to impart novel physical properties and biological roles to proteins from both eukaryotes and prokaryotes. In this study, gel-based glycoproteomics were used to identify glycoproteins of the potential biothreat agent Burkholderia pseudomallei and the closely related but nonpathogenic B. thailandensis. Top-down and bottom-up mass spectrometry (MS) analyses identified that the flagellin proteins of both species were posttranslationally modified by novel glycans. Analysis of proteins from two strains of each species demonstrated that B. pseudomallei flagellin proteins were modified with a glycan with a mass of 291 Da, while B. thailandensis flagellin protein was modified with related glycans with a mass of 300 or 342 Da. Structural characterization of the B. thailandensis carbohydrate moiety suggests that it is an acetylated hexuronic acid. In addition, we have identified through mutagenesis a gene from the lipopolysaccharide (LPS) O-antigen biosynthetic cluster which is involved in flagellar glycosylation, and inactivation of this gene eliminates flagellar glycosylation and motility in B. pseudomallei. This is the first report to conclusively demonstrate the presence of a carbohydrate covalently linked to a protein in B. pseudomallei and B. thailandensis, and it suggests new avenues to explore in order to examine the marked differences in virulence between these two species.

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Section: Discussionmentioning

confidence: 99%

Flagellar Glycosylation in Burkholderia pseudomallei and Burkholderia thailandensis

Scott¹,

Twine²,

Fulton³

et al. 2011

Journal of Bacteriology

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“…The bacteria used in this study were B. pseudomallei strains K96243 (34), RR2808 (⌬purM ⌬wcbB) (35), and RR2683 (⌬purM ⌬rmlD) (19) and Escherichia coli BL21 Star(DE3) (pLysS)(pCRT7/NT-TOPO-LolC) (36). B. pseudomallei strains RR2808 and RR2683 are derivatives of the adenine auxotroph Bp82 (37), a selectagent-excluded ⌬purM derivative of strain 1026b.…”

Section: Methodsmentioning

confidence: 99%

Burkholderia pseudomallei Capsular Polysaccharide Conjugates Provide Protection against Acute Melioidosis

et al. 2014

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bBurkholderia pseudomallei, the etiologic agent of melioidosis, is a CDC tier 1 select agent that causes severe disease in both humans and animals. Diagnosis and treatment of melioidosis can be challenging, and in the absence of optimal chemotherapeutic intervention, acute disease is frequently fatal. Melioidosis is an emerging infectious disease for which there are currently no licensed vaccines. Due to the potential malicious use of B. pseudomallei as well as its impact on public health in regions where the disease is endemic, there is significant interest in developing vaccines for immunization against this disease. In the present study, type A O-polysaccharide (OPS) and manno-heptose capsular polysaccharide (CPS) antigens were isolated from nonpathogenic, select-agent-excluded strains of B. pseudomallei and covalently linked to carrier proteins. By using these conjugates (OPS2B1 and CPS2B1, respectively), it was shown that although high-titer IgG responses against the OPS or CPS component of the glycoconjugates could be raised in BALB/c mice, only those animals immunized with CPS2B1 were protected against intraperitoneal challenge with B. pseudomallei. Extending upon these studies, it was also demonstrated that when the mice were immunized with a combination of CPS2B1 and recombinant B. pseudomallei LolC, rather than with CPS2B1 or LolC individually, they exhibited higher survival rates when challenged with a lethal dose of B. pseudomallei. Collectively, these results suggest that CPSbased glycoconjugates are promising candidates for the development of subunit vaccines for immunization against melioidosis.

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“…Subunit vaccines have also been developed for immunization against B. pseudomallei, with the most promising protection to date being achieved with purified proteins such as BipD (34) and BipB and BipC (36); with recombinant proteins, including Hcp1, Hcp2, Hcp3, and Hcp6 (37); with lipopolysaccharide (LPS) (38); with purified flagellin (39); with LolC, PotF, and OppA nonmembrane protein (40); with outer membrane vesicles (41); and with recombinant Omp85 protein (42). However, the use of single antigen subunit vaccines for Burkholderia infection is unlikely to generate broad protective immunity against this very genetically diverse and unstable organism (35,43).…”

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confidence: 99%

Correlates of Immune Protection following Cutaneous Immunization with an Attenuated Burkholderia pseudomallei Vaccine

et al. 2013

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Infections with the Gram-negative bacterium Burkholderia pseudomallei (melioidosis) are associated with high mortality, and there is currently no approved vaccine to prevent the development of melioidosis in humans. Infected patients also do not develop protective immunity to reinfection, and some individuals will develop chronic, subclinical infections with B. pseudomallei. At present, our understanding of what constitutes effective protective immunity against B. pseudomallei infection remains incomplete. Therefore, we conducted a study to elucidate immune correlates of vaccine-induced protective immunity against acute B. pseudomallei infection. BALB/c and C57BL/6 mice were immunized subcutaneously with a highly attenuated, Select Agent-excluded purM deletion mutant of B. pseudomallei (strain Bp82) and then subjected to intranasal challenge with virulent B. pseudomallei strain 1026b. Immunization with Bp82 generated significant protection from challenge with B. pseudomallei, and protection was associated with a significant reduction in bacterial burden in lungs, liver, and spleen of immunized mice. Humoral immunity was critically important for vaccine-induced protection, as mice lacking B cells were not protected by immunization and serum from Bp82-vaccinated mice could transfer partial protection to nonvaccinated animals. In contrast, vaccineinduced protective immunity was found to be independent of both CD4 and CD8 T cells. Tracking studies demonstrated uptake of the Bp82 vaccine strain predominately by neutrophils in vaccine-draining lymph nodes and by smaller numbers of dendritic cells (DC) and monocytes. We concluded that protection following cutaneous immunization with a live attenuated Burkholderia vaccine strain was dependent primarily on generation of effective humoral immune responses.

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Identification of a LolC Homologue inBurkholderia pseudomallei, a Novel Protective Antigen for Melioidosis

Abstract: Melioidosis is an emerging disease of humans in

Cited by 61 publications

References 40 publications

Flagellar Glycosylation in Burkholderia pseudomallei and Burkholderia thailandensis

Flagellar Glycosylation in Burkholderia pseudomallei and Burkholderia thailandensis

Burkholderia pseudomallei Capsular Polysaccharide Conjugates Provide Protection against Acute Melioidosis

Correlates of Immune Protection following Cutaneous Immunization with an Attenuated Burkholderia pseudomallei Vaccine

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