Genetically detoxified pertussis toxin displays near identical structure to its wild-type and exhibits robust immunogenicity

Abstract

The mutant gdPT R9K/E129G is a genetically detoxified variant of the pertussis toxin (PTx) and represents an attractive candidate for the development of improved pertussis vaccines. The impact of the mutations on the overall protein structure and its immunogenicity has remained elusive. Here we present the crystal structure of gdPT and show that it is nearly identical to that of PTx. Hydrogen-deuterium exchange mass spectrometry revealed dynamic changes in the catalytic domain that directly impacted NAD(+)binding which was confirmed by biolayer interferometry. Distal changes in dynamics were also detected in S2-S5 subunit interactions resulting in tighter packing of B-oligomer corresponding to increased thermal stability. Finally, antigen stimulation of human whole blood, analyzed by a previously unreported mass cytometry assay, indicated broader immunogenicity of gdPT compared to pertussis toxoid. These findings establish a direct link between the conserved structure of gdPT and its ability to generate a robust immune response. Ausar et al. show that the crystal structure of gdPT, a mutant pertussis toxin (PTx), is nearly identical to that of PTx. They also find that gdPT exhibits broader immunogenicity than the pertussis toxoid antigen. This study suggests a promising potential of gdPT as an improved acellular pertussis vaccine candidate due to its reduced toxicity and greater immunogenicity.