Exploiting the Burkholderia pseudomallei Acute Phase Antigen BPSL2765 for Structure-Based Epitope Discovery/Design in Structural Vaccinology

Abstract

We solved the crystal structure of Burkholderia pseudomallei acute phase antigen BPSL2765 in the context of a structural vaccinology study, in the area of melioidosis vaccine development. Based on the structure, we applied a recently developed method for epitope design that combines computational epitope predictions with in vitro mapping experiments and successfully identified a consensus sequence within the antigen that, when engineered as a synthetic peptide, was selectively immunorecognized to the same extent as the recombinant protein in sera from melioidosis-affected subjects. Antibodies raised against the consensus peptide were successfully tested in opsonization bacterial killing experiments and antibody-dependent agglutination tests of B. pseudomallei. Our strategy represents a step in the development of immunodiagnostics, in the production of specific antibodies and in the optimization of antigens for vaccine development, starting from structural and physicochemical principles.

Effect of Host Factors on Neutrophil Functions in Response to Burkholderia pseudomallei in Healthy Thai Subjects

“…This result is consistent with our previous studies, which showed that levels of antibody against BPSL2765 (OmpA protein) were 10-fold higher in individuals with melioidosis (12). A recent study showed that the levels of specific antibodies that increased against the BPSL2765 peptide effectively enhanced phagocytosis and ROS production in an opsonization and killing assay using B. pseudomallei (16). This result suggests that the levels of antibodies are likely to play an important role in the response to B. pseudomallei infection.…”

Structure-based Epitope Mapping of Mycobacterium tuberculosis Secretary Antigen MTC28

“…Linear epitopes sometimes have an elevated level of antibody reactivity than the parental antigen. As for example, two acute phase antigens (BPSL 2765 and OppA) of Burkholderia pseudomallei (46,47). An antigen contains several epitopes, some of which are specific to the parental organism.…”

Boosting of post‐exposure human T‐cell and B‐cell recall responses in vivo by Burkholderia pseudomallei‐related proteins

“…17,40,41 Lipopolysaccharide removal was carried out by incubating each purified protein sample overnight at 4°, in a Pierce High Capacity Endotoxin Removal Spin Column (Thermo Fisher Scientific, MA), according to the manufacturer's instructions. 17,40,41 Lipopolysaccharide removal was carried out by incubating each purified protein sample overnight at 4°, in a Pierce High Capacity Endotoxin Removal Spin Column (Thermo Fisher Scientific, MA), according to the manufacturer's instructions.…”