A recombinant hybrid outer membrane protein for vaccination against Pseudomonas aeruginosa
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Why a vaccine against Pseudomonas aeruginosa?
P. aeruginosa is an opportunistic pathogen which infects cystic fibrosis patients and represents a major risk among hospital-acquired infections, especially in burn and otherwise immunocompromized patients, including transplantation and cancer patients. Because of the wide distribution of natural and induced resistance to P. aeruginosa the chemotherapeutical possibilities remain restricted to only a few and partly toxic drugs.
Due to the uncontrolled use of antibacterial chemotherapeutics and
Targets for the development of a vaccine
There are varying approaches when developing vaccines against P. aeruginosa depending on the bacterial target used: flagella, pili, alginate, lipopolysaccharides or the outer membrane proteins. As the outer membrane proteins have been shown to be antigenically crossreactive within different serogroups and clinical isolates[2], they were chosen as vaccine candidates.
We focused our efforts on the two outer membrane proteins OprI[3]and OprF[4]of P. aeruginosa. OprI is a lipoprotein which contains
Construction and testing of a hybrid antigen
We combined the segment from aa 190 to aa 342 of OprF with the mature OprI protein because such a hybrid antigen based on two different outer membrane proteins has the advantage of producing one single recombinant antigen which is more cost-effective than the development and production of two different recombinant proteins. The hybrid protein was shown to be efficacious in two different animal models, a model for active and a model for passive immunization.
For active immunization, the hybrid
Clinical pilot study with the hybrid antigen
A purification procedure for the recombinant OprF/OprI hybrid antigen was established. After rupture of the E. coli cells the supernatant was loaded onto a metal chelate column. The His-tagged recombinant protein was eluted by imidazole which was removed by a gel filtration step. The LPS was separated from the product by acid precipitation. The LPS content was shown to be lower than 30 EU/mg protein and the specific purity was determined to be ≥98%.
A clinical pilot study was performed using the
Antibody response against OprF and OprI in P. aeruginosa infected patients
Our proposed target group for the clinical testing of our hybrid antigen vaccine are burn patients known to have an incidence of 35–50% infection by P. aeruginosa. We have analyzed sera of infected burn patients and found that they develop antibodies against OprF and OprI, the titer correlating with the total burn surface and the infection time. We conclude that burn patients are able to form antibodies against OprF and OprI. Therefore vaccination of burn patients with the hybrid antigen should
Future directions
The next step in the direction of vaccine development is the testing of safety and immunogenicity of the hybrid antigen in burn patients. A prerequisite for the protection of this patient group against an infection with P. aeruginosa is a good immunological response to the vaccine after just one single immunization or at the latest after two short time immunizations. This is being tested in a further clinical pilot study including burn patients.
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Cited by (31)
Immunization with outer membrane proteins (OprF and OprI) and flagellin B protects mice from pulmonary infection with mucoid and nonmucoid Pseudomonas aeruginosa
2018, Journal of Microbiology, Immunology and InfectionCitation Excerpt :The combined vaccine showed a significant protective effect (Figure 4A and B). This protection could be explained by synergistic effect of both antibodies that binds to different epitopes on the surface of the bacteria.30 Vaccines consisting of a mixture of different outer membrane proteins or fusion components of these proteins could increase the degree of protection against pseudomonas infection.31
Anti-Pseudomonas aeruginosa IgY antibodies augment bacterial clearance in a murine pneumonia model
2016, Journal of Cystic FibrosisCitation Excerpt :The efficacy of active immunization to reduce P. aeruginosa lung infection has been evaluated in vivo with some attainment. Conjugate vaccines targeting P. aeruginosa polysaccharides have been successfully tested in animal models [21,22] and several other bacterial antigens like flagella [23], pili [24] and outer membrane proteins [25] encompass immunogenic potentials to induce a protective immune response in vivo. However, the current accessible clinical vaccination studies have failed to authenticate any convincing validity of P. aeruginosa-specific vaccines in CF [26–28].
Vaccination efficiency of surface antigens and killed whole cell of Pseudomonas putida in large yellow croaker (Pseudosciaena crocea)
2013, Fish and Shellfish ImmunologyCitation Excerpt :In P. aeruginosa, LPS plays a key role in pathogenesis and the O-antigen is a major target for protective immunity [20,21]. Some major outer membrane proteins, such as OprF and OprI, are highly conserved [22], and targeted vaccines have provided efficient protection in most experiments [23–26]. Pseudomonads usually produce an EPS matrix to form a biofilm community [27,28].