Titolo della tesi: In vitro spatial and temporal dynamics of Gonococcus interaction to host cells: Time resolving Dual RNA-Seq
Neisseria gonorrhoeae is the leading cause of gonorrhea, a sexually transmitted infection representing a major public health concern globally due to the high rise of untreatable infections and to the high resistance to antibiotics, raising the urgent need of new preventive countermeasures. Neisseria gonorrhoeae is an exclusive human pathogen having the human genital tracts as initial sites of infection. We analyzed the temporal and spatial adhesion/internalization of Neisseria gonorrhoeae strains to three human cell models (cervix, urethra and oropharynx), exploiting an innovative technology based on the measurement of labeled bacteria by a high content screening (HCS) fluorescence microscope platform, Opera Phenix. This analysis has highlighted that the gonococcal interaction with the host cells seems to be strain dependent and cell line dependent, raising interest on mechanisms guiding the pathogenic process in the various anatomical districts.
The complex interplay between infecting bacteria and the human host is key for the understanding of pathogenesis and disease progression and for the identification of crucial virulence determinants. The Dual RNA seq approach allows the monitoring of genes from both host and pathogen and it is a powerful method to investigate the infection dynamics. Our work highlights the phenotypical and transcriptional changes, in a time course of infection, from initial contact to invasion, using relevant in vitro models that mimic the different anatomical sites encountered by the pathogen during the natural infection.