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Published on: January 8, 2020
Evaluation of outer membrane proteins of Pseudomonas aeruginosa as a protective agent in mice model
Mohammad Kazem Sharifi-Yyazdi1, Farhad Esmaily, Farhad Vaezzadeh
1Department of Medical Laboratory Sciences, Faculty of Para medicine, Tehran Medical Sciences, Tehran, Iran.
Abstract:
The crude Outer Membrane Protein (OMP) from a strain of P. aeruginosa isolated from burn patient was purified by two different methods. One procedure involved separation of Sodium Dodecyle Sulphate (SDS) and Triton X-100, where as the other involved using lysozyme enzyme. Both methods showed very similar polypeptide pattern and the major peptide band with molecular weight of 37 KD was common in both procedures. The protein estimation of OMP extracted by lysosyme was 3 mg mL(-1) compared to 5.5 mg mL(-1) extracted by Triton-X100 method. The latter was chosen to examine for the immunogenicity study in a mice model. The efficacy of immunization with OMP and challenge with homologous strain in mice showed a very good protection compared to control mice injected with saline. The passive haemoagglutination test (PHA) in mice, injected with OMP showed increased level of antibody after the second injection and stayed constant after repeated injection. The results of this study showed that the crude OMP extracted from P. aeruginosa induced a significant protection in mice against Pseudomonas infections and could be used as a vaccine candidate.
Insights
Outer Membrane Protein (OMP) from Pseudomonas aeruginosa, a burn patient isolate, was purified. Immunization with OMP provided significant protection in mice against Pseudomonas infections, indicating its potential as a vaccine candidate.
Area of Science:
- Microbiology
- Immunology
- Biochemistry
Background:
- Pseudomonas aeruginosa is a significant opportunistic pathogen, particularly in burn wound infections.
- Outer Membrane Proteins (OMPs) are key targets for developing vaccines against bacterial infections.
Purpose of the Study:
- To purify Outer Membrane Protein (OMP) from a P. aeruginosa strain isolated from a burn patient.
- To evaluate the immunogenicity and protective efficacy of the purified OMP in a murine model.
Main Methods:
- OMP purification using Sodium Dodecyle Sulphate (SDS) and Triton X-100, and lysozyme enzyme.
- Protein estimation and polypeptide pattern analysis.
- Immunization of mice with OMP, followed by challenge with homologous P. aeruginosa strain and passive haemoagglutination (PHA) test.
Main Results:
- Two purification methods yielded similar polypeptide patterns, with a common 37 kDa peptide band.
- Triton X-100 method yielded higher protein concentration (5.5 mg/mL) compared to lysozyme (3 mg/mL).
- OMP immunization conferred significant protection against P. aeruginosa challenge and increased antibody levels in mice.
Conclusions:
- Crude OMP from P. aeruginosa is immunogenic and protective in a mouse model.
- The purified OMP demonstrates potential as a vaccine candidate for preventing Pseudomonas infections.
