Background Thiamine pyrophosphate (TPP) is an important metabolite that affects many metabolic pathways within the cell.) Thiamine (thi) auxotrophs of Neisseria gonorrhoeae (Gc) that could not grow without TPP or other thiamine derivatives were isolated from patients in the 1970’s, but the effect of thi auxotrophy on Gc pathogenesis is not known. We recently demonstrated that a genetically defined Gc mutant that cannot biosynthesize TPP is more susceptible to killing by neutrophils, cationic antimicrobial peptides (CAMPs), and reactive oxygen species (ROS) in vitro, and attenuated for experimental infection of mice that have a neutrophil response to infection. Here we investigated the susceptibility of five recently isolated TPP auxotrophs to paraquat, an inducer of ROS and CAMPs as a first step towards understanding the consequence of thi auxotrophy during human infection.
Methods Eighty-nine Gc isolates in the USUHS Gc Resistance and Reference Repository isolated between 2014 and 2017 were screened for the capacity to grow on medium without TPP. Auxotrophs were tested for susceptibility to paraquat and colistin (polymyxin E) using standard methods.
Results Five thi auxotrophs were identified among the 89 isolates tested (5.6%). Four of the auxotrophs exhibited increased susceptibility to paraquat and colistin compared to a wild-type Gc strain. Auxotroph 4097, in contrast, showed a ∼2-fold greater resistance to 0.0195 mM of paraquat and colistin compared to a wild-type Gc strain, suggesting this isolate may carry a compensatory mutation(s).
Conclusion We conclude that clinically isolated thi auxotrophs are more susceptible to ROS and CAMPS. We hypothesize that these isolates may have a lesser ability to withstand oxygen-dependent and independent effectors of the host inflammatory response and that selection for compensatory mutations during infection may be one mechanism by which thi auxotrophs remain in circulation. Analysis of WGS data is underway to identify the genetic basis of thi auxotrophy and possible compensatory mutations.
Disclosure No significant relationships.t
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.