Immunological properties and protective efficacy of a single mycobacterial antigen displayed on polyhydroxybutyrate beads

Rubio‐Reyes, Patricia; Parlane, Natalie A.; Buddle, Bryce M.; Wedlock, D. Neil; Rehm, Bernd H. A.

doi:10.1111/1751-7915.12754

Cited by 10 publications

(10 citation statements)

References 28 publications

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“…2B). The polyester synthase PhaC remained anchored on the surface after particle formation and isolation (29,39). Protein profile analysis by SDS-PAGE showed that the full-length PhaC-TB antigen fusions were overproduced because the MWs of fusion proteins corresponded to their theoretical MWs (Fig.…”

Section: Discussionmentioning

confidence: 99%

Innovative antigen carrier system for the development of tuberculosis vaccines

et al. 2019

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A major obstacle to tuberculosis (TB)‐subunit–vaccine development has been the induction of inadequate levels of protective immunity due to the limited breadth of antigen in vaccine preparations. In this study, immunogenic mycobacterial fusion peptides Ag85B‐TB10.4 and Ag85B‐TB10.4‐Rv2660c were covalently displayed on the surface of self‐assembled polyester particles. This study investigated whether polyester particles displaying mycobacterial antigens could provide augmented immunogenicity (i.e., offer an innovative vaccine formulation) when compared with free soluble antigens. Herein, polyester particle–based particulate vaccines were produced in an endotoxin‐free Escherichia coli strain and emulsified with the adjuvant dimethyl dioctadecyl ammonium bromide. C57BL/6 mice were used to study the immunogenicity of formulated particulate vaccines. The result of humoral immunity showed the antibodies only interacted with target antigens and not with PhaC and the background proteins of the production host. The analysis of T helper 1 cellular immunity indicated that a relatively strong production of cellular immunity biomarkers, IFN‐γ and IL‐17A cytokines, was induced by particulate vaccines when compared with the respective soluble controls. This study demonstrated that polyester particles have the potential to perform as a mycobacterial antigen–delivery agent to induce augmented antigen‐specific immune responses in contrast to free soluble vaccines.—Chen, S., Sandford, S., Kirman, J. R., Rehm, B. H. A. Innovative antigen carrier system for the development of tuberculosis vaccines. FASEB J. 33, 7505–7518 (2019). http://www.fasebj.org

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Section: Discussionmentioning

confidence: 99%

Innovative antigen carrier system for the development of tuberculosis vaccines

et al. 2019

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“…Nevertheless, the construction of bioactive PHA systems by protein immobilization has been investigated to a limited degree at the laboratory scale and is still a nascent technology that needs scaling up for industrial applications (43). Very efficient covalent methods for PHA functionalization-based translational fusions with PhaC from Ralstonia eutropha have been described in detail by Rehm's group, leading to a variety of biotechnological and biomedical applications (3,44,45). However, in the case of noncovalent methods like those based in phasins, several key technical issues, such as control of PHA-protein association/dissociation and granule size, still remain unresolved.…”

Section: Discussionmentioning

confidence: 99%

Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins

Bello-Gil

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Fonseca

et al. 2018

Appl Environ Microbiol

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Polyhydroxyalkanoates (PHAs) are biodegradable polyesters that accumulate in the cytoplasm of certain bacteria. One promising biotechnological application utilizes these biopolymers as supports for protein immobilization. Here, the PHA-binding domain of the KT2440 PhaF phasin (BioF polypeptide) was investigated as an affinity tag for the functionalization of poly-3-hydroxybutyrate (PHB) particles with recombinant proteins, namely, full-length PhaF and two fusion proteins tagged to BioF (BioF-C-LytA and BioF-β-galactosidase, containing the choline-binding module C-LytA and the β-galactosidase enzyme, respectively). The protein-biopolyester interaction was strong and stable at a wide range of pHs and temperatures, and the bound protein was highly protected from self-degradation, while the binding strength could be modulated by coating with amphiphilic compounds. Finally, BioF-β-galactosidase displayed very stable enzymatic activity after several continuous activity-plus-washing cycles when immobilized in a minibioreactor. Our results demonstrate the potentialities of PHA and the BioF tag for the construction of novel bioactive materials. Our results confirm the biotechnological potential of the BioF affinity tag as a versatile tool for functionalizing PHA supports with recombinant proteins, leading to novel bioactive materials. The wide substrate range of the BioF tag presumably enables protein immobilization of virtually all natural PHAs as well as blends, copolymers, or artificial chemically modified derivatives with novel physicochemical properties. Moreover, the strength of protein adsorption may be easily modulated by varying the coating of the support, providing new perspectives for the engineering of bioactive materials that require a tight control of protein loading.

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“…Moreover, IL17A, IFNγ, and IL2 are known to be important for protection against TB; however, their production postvaccination does not always correlate tightly with enhanced protective immunity. [32b,37] Future animal studies including challenge with the pathogen will investigate whether the induced immune responses prevent infection by M. tuberculosis .…”

Section: Resultsmentioning

confidence: 99%

Design of Bacterial Inclusion Bodies as Antigen Carrier Systems

et al. 2018

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Synthetic subunit vaccines hold great promise to prevent infectious diseases, but they often induce only weak immune responses. Here bioengineering techniques to develop immunogenic antigen particles comprised of self‐assembling antigenic peptides are exploited. A bioparticle platform is developed to present repetitive copies of subunit antigens, which mimic the host‐pathogen surface interaction to provide enhanced immunogenicity. Herein, mycobacterial fusion proteins H4 (consisting of Ag88B‐TB10.4) and H28 (containing Ag88B‐TB10.4‐Rv2660c) are bioengineered to assemble H4 and/or H28 antigens into particulate vaccines inside an endotoxin‐free Escherichia coli strain, to serve as a novel vaccine against tuberculosis (TB). A bioprocess for particle isolation is developed. The physicochemical properties of the particles are investigated. Antigen particles formulated with the adjuvant dimethyl dioctadecyl ammonium bromide are used to subcutaneously immunize C57BL/6 mice. Both soluble and particulate vaccines elicit production of functional and specific antibodies, which only interact with target antigens and not production host cell proteins. Particulate vaccines induce strong production of interferon gamma (IFNγ) and interleukin 17A (IL17A) cytokines. When compared to soluble antigens, the antigen particle vaccines show increased immunogenicity. Overall, this study provides proof of concept that selected antigens can be engineered to self‐assemble into inclusion bodies to serve as particulate vaccines with improved immunological properties compared to the respective soluble antigens.

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Immunological properties and protective efficacy of a single mycobacterial antigen displayed on polyhydroxybutyrate beads

Cited by 10 publications

References 28 publications

Innovative antigen carrier system for the development of tuberculosis vaccines

Innovative antigen carrier system for the development of tuberculosis vaccines

Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins

Design of Bacterial Inclusion Bodies as Antigen Carrier Systems

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