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Review
. 2021 Jan 22;13(2):83.
doi: 10.3390/toxins13020083.

Bioengineering of Bordetella pertussis Adenylate Cyclase Toxin for Vaccine Development and Other Biotechnological Purposes

Daniel Ladant  1
Affiliations
Review

Bioengineering of Bordetella pertussis Adenylate Cyclase Toxin for Vaccine Development and Other Biotechnological Purposes

Daniel Ladant. Toxins (Basel). .

Abstract

The adenylate cyclase toxin, CyaA, is one of the key virulent factors produced by Bordetella pertussis, the causative agent of whooping cough. This toxin primarily targets innate immunity to facilitate bacterial colonization of the respiratory tract. CyaA exhibits several remarkable characteristics that have been exploited for various applications in vaccinology and other biotechnological purposes. CyaA has been engineered as a potent vaccine vehicle to deliver antigens into antigen-presenting cells, while the adenylate cyclase catalytic domain has been used to design a robust genetic assay for monitoring protein-protein interactions in bacteria. These two biotechnological applications are briefly summarized in this chapter.

Keywords: Bordetella pertussis; adenylate cyclase; antigen delivery; biological screening; protein–protein interactions; recombinant toxin; recombinant vaccine; two-hybrid.

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Conflict of interest statement

D.L. is the co-inventor of patents protecting the use of recombinant CyaA as an antigen delivery vector and bacterial two-hybrid system.

Figures

Figure 1
Figure 1
Schematic representation of CyaA structural organization and biogenesis. Upper part: Structural organization of the CyaA polypeptide with its different subdomains. Lower part left: in B. pertussis, CyaA is synthesized as an inactive precursor proCyaA that is activated by the CyaC acyltransferase and then secreted across the bacterial envelope by a dedicated type I secretion system (T1SS); Lower part right: model for CyaA invasion of eukaryotic cells. After binding to CD11b/CD18 receptor (j), CyaA inserts its hydrophobic segments into the membrane and delivers its catalytic domain into the cytosol where it is activated by calmodulin (CaM) to overproduce cAMP.
Figure 2
Figure 2
Recombinant CyaA for antigen delivery into dendritic cells (DCs). An antigen of interest (orange) is genetically inserted into one of the permissive sites (yellow) located within the CyaA catalytic domain. When injected into an animal, the purified CyaA-antigen fusion binds to the CD11b/CD18 receptor (j) at the surface of the dendritic cells. Upon translocation of the AC domain across the plasma membrane, the antigen is processed by proteasome and enters the MHC-I presentation pathway (via the endoplasmic reticulum, ER) leading to the activation of cytotoxic CD8+ T-cells. Alternatively, the recombinant CyaA is endocytosed, and the grafted antigen enters the endosomal pathway for MHC-II presentation, leading to activation of specific CD4+ helper T-cells.
Figure 3
Figure 3
Principle of BACTH assay. (A) In an E. coli cya strain, the T25 and T18 fragments of CyaA are co-expressed as fusions with polypeptides X and Y; interaction between X and Y triggers heterodimerization of the hybrid proteins leading to cAMP synthesis. (B) The catabolite activator protein (CAP) binds cAMP and activates transcription of catabolic operons (reporter gene). (C) Diverse topological arrangement and subcellular localization of hybrid proteins; “im” represents the bacterial inner membrane.
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