Impact of Detergent on Biophysical Properties and Immune Response of the IpaDB Fusion Protein, a Candidate Subunit Vaccine against Shigella Species

Shigellaspp. are causative agents of bacillary dysentery, a human illness with high global morbidity levels, particularly among elderly and infant populations.Shigellainfects via the fecal-oral route, and its virulence is dependent upon a type III secretion system (T3SS). Two components of the exposed needle tip complex of theShigellaT3SS, invasion plasmid antigen D (IpaD) and IpaB, have been identified as broadly protective antigens in the mouse lethal pneumonia model. A recombinant fusion protein (DB fusion) was created by joining the coding sequences of IpaD and IpaB. The DB fusion is coexpressed with IpaB's cognate chaperone, IpgC, for proper recombinant expression. The chaperone can then be removed by using the mild detergents octyl oligooxyethelene (OPOE) orN,N-dimethyldodecylamineN-oxide (LDAO). The DB fusion in OPOE or LDAO was used for biophysical characterization and subsequent construction of an empirical phase diagram (EPD). The EPD showed that the DB fusion in OPOE is most stable at neutral pH below 55°C. In contrast, the DB fusion in LDAO exhibited remarkable thermal plasticity, since this detergent prevents the loss of secondary and tertiary structures after thermal unfolding at 90°C, as well as preventing thermally induced aggregation. Moreover, the DB fusion in LDAO induced higher interleukin-17 secretion and provided a higher protective efficacy in a mouse challenge model than did the DB fusion in OPOE. These data indicate that LDAO might introduce plasticity to the protein, promoting thermal resilience and enhanced protective efficacy, which may be important in its use as a subunit vaccine.

A Molecular Dynamics Study of the Terminal Zr(Ni) Solid Solutions

The Zr terminal portion of the Zr-Ni phase diagram has been evaluated by means of a many-body tight-binding potential. The internal energy curves of the Zr(Ni) solid solutions at T=300 K have been calculated by Molecular Dynamics simulations. These curves exhibit positive values, contrary to former empirical phase diagram calculations. Implications of these results relevant to the problem of amorphization in metallic systems by solid-state reactions are discussed.