title: Impact of detergent on biophysical properties and immune response of the IpaDB fusion protein, a candidate subunit vaccine against Shigella species.
abstract: Shigella spp. are causative agents of bacillary dysentery, a human illness with high global morbidity levels, particularly among elderly and infant populations. Shigella infects 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 the Shigella T3SS, 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) or N,N-dimethyldodecylamine N-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.
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
authors.forename: Xiaotong
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
authors.forename: Shyamal P
authors.initials: SP
authors.lastname: Choudhari
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Martinez-Becerra
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA.
authors.forename: Jae Hyun
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Dickenson
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA.
authors.forename: Ronald T
authors.initials: RT
authors.lastname: Toth
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA.
authors.forename: Sangeeta B
authors.initials: SB
authors.lastname: Joshi
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
authors.forename: Jamie C
authors.initials: JC
authors.lastname: Greenwood
authors.affiliation: Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, Louisiana, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Clements
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
authors.forename: William D
authors.initials: WD
authors.lastname: Picking
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA.
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA wendy.picking@ku.edu.
authors.forename: Wendy L
authors.initials: WL
authors.lastname: Picking
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry* / Q000276:immunology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000276:immunology*; D001428:Bacterial Vaccines / Q000737:chemistry* / Q000276:immunology*; D055598:Chemical Phenomena / Q000187:drug effects; D003902:Detergents / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D051379:Mice; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology; D013696:Temperature
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D003902:Detergents; C095770:IpaD protein, Shigella flexneri; D011993:Recombinant Fusion Proteins; C087447:ipaB protein, Shigella