Pseudomonas aeruginosa is a dominant pathogen in people with cystic fibrosis (CF) contributing to morbidity and mortality. Its tremendous ability to adapt greatly facilitates its capacity to cause chronic infections. The adaptability and flexibility of the pathogen are afforded by the extensive number of virulence factors it has at its disposal, providing P. aeruginosa with the facility to tailor its response against the different stressors in the environment. A deep understanding of these virulence mechanisms is crucial for the design of therapeutic strategies and vaccines against this multi-resistant pathogen. Therefore, this review describes the main virulence factors of P. aeruginosa and the adaptations it undergoes to persist in hostile environments such as the CF respiratory tract. The very large P. aeruginosa genome (5 to 7 MB) contributes considerably to its adaptive capacity; consequently, genomic studies have provided significant insights into elucidating P. aeruginosa evolution and its interactions with the host throughout the course of infection.

Pseudomonas aeruginosa is a leading cause of chronic respiratory infections in people with cystic fibrosis (CF), bronchiectasis or chronic obstructive pulmonary disease (COPD), and acute infections in immunocompromised individuals. The adaptability of this opportunistic pathogen has hampered the development of antimicrobial therapies, and consequently, it remains a major threat to public health. Due to its antimicrobial resistance, vaccines represent an alternative strategy to tackle the pathogen, yet despite over 50 years of research on anti-Pseudomonas vaccines, no vaccine has been licensed. Nevertheless, there have been many advances in this field, including a better understanding of the host immune response and the biology of P. aeruginosa. Multiple antigens and adjuvants have been investigated with varying results. Although the most effective protective response remains to be established, it is clear that a polarised Th2 response is sub-optimal, and a mixed Th1/Th2 or Th1/Th17 response appears beneficial. This comprehensive review collates the current understanding of the complexities of P. aeruginosa-host interactions and its implication in vaccine design, with a view to understanding the current state of Pseudomonal vaccine development and the direction of future efforts. It highlights the importance of the incorporation of appropriate adjuvants to the protective antigen to yield optimal protection.

Antimicrobial-resistant pathogens such as Pseudomonas aeruginosa and Acinetobacter baumannii can cause potentially fatal infections in susceptible individuals, with respiratory tract infections among the most common clinical presentations. The development of novel treatments or prophylactic interventions to combat these infections is urgently needed and requires robust, reliable animal models for their preclinical evaluation. In particular, the bacterial burden needs to be accurately determined before and after administration of the potential therapy under evaluation to quantify the effectiveness of the treatment. We provide two reliable, non-invasive murine acute lung challenge models with either P. aeruginosa or A. baumannii using an oropharyngeal aspiration technique, which has been widely overlooked in studies testing vaccines or treatments for these pathogens. Here, we show that this non-surgical technique to deliver suspensions into mouse lungs does not significantly impact animal welfare (based on welfare monitoring and weight) and allows uniform bilateral distribution of the bacterial dose, resulting in even bioburden in both lungs. The optimal timepoint for humane killing and organ harvest was 24 h after challenge for both pathogens, and at least 4×10⁶ and 10⁷ c.f.u. per mouse were needed to obtain a reproducible P. aeruginosa or A. baumannii bioburden, respectively. These mouse challenge models offer a valuable tool to assess therapeutic interventions against P. aeruginosa or A. baumannii infections.

Antimicrobial resistant pathogens such as Pseudomonas aeruginosa and Acinetobacter baumannii can cause potentially fatal infections in susceptible individuals, with respiratory tract infections among the most common clinical presentation. The development of novel treatments or prophylactic interventions to combat these infections is urgently needed, and requires robust, reliable animal models for their preclinical evaluation. In particular, the bacterial burden needs to be accurately determined before and after administration of the potential therapy under evaluation to quantitate the effectiveness of the treatment. We provide two reliable, non-invasive murine acute lung challenge models with either P. aeruginosa or A. baumannii using an oropharyngeal aspiration technique, which has been widely overlooked in studies testing vaccines or treatments for these pathogens. Here, we show that this non-surgical technique to deliver suspensions into mouse lungs does not significantly impact animal welfare (based on welfare monitoring and weight) and allows uniform bilateral distribution of the bacterial dose resulting in even bioburden in both lungs. The optimal timepoint for humane killing and organ harvest was 24 h after challenge for both pathogens, and at least 4x106 and 107 CFU/mouse were needed to obtain a reproducible P. aeruginosa or A. baumannii bioburden, respectively. These mouse challenge models offer a valuable tool to assess therapeutic interventions against P. aeruginosa or A. baumannii infections.

Bacterial infections caused by antimicrobial resistant pathogens such as Pseudomonas aeruginosa and Acinetobacter baumannii can cause potentially fatal infections in susceptible individuals, with respiratory tract infections among the most common presentation. There is a real need to develop novel treatments or prophylactic interventions to combat these infections. The development of any new treatment or vaccine requires robust, reliable animal models for their preclinical evaluation. In particular, the bacterial burden needs to be accurately determined before and after administration of the potential therapy under evaluation to quantitate the effectiveness of the treatment. We provide a reliable non-invasive murine model of acute lung infection with either P. aeruginosa or A. baumannii using an oropharyngeal aspiration technique. This non-surgical technique to deliver suspensions into mouse lungs is less stressful to animals than other techniques. It allowed uniform bilateral distribution of the bacterial dose resulting in even colonisation of both lungs. The optimal timepoint for humane killing and organ harvest was 24 h after challenge for both pathogens, and at least 4x106 and 107 CFU/mouse were needed to establish P. aeruginosa or A. baumannii infection, respectively. These optimised mouse pneumonia models offer a valuable tool for studying P. aeruginosa and A. baumannii infections and evaluating therapeutic interventions against them.