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Published on: October 26, 2016
Synthesis and characterization of Pseudomonas aeruginosa alginate-tetanus toxoid conjugate
Nasim Kashef, Qorban Behzadian-Nejad, Shahin Najar-Peerayeh
1Iranian Blood Transfusion Organization, Research Center, Tehran, Iran.
Abstract:
Chronic infection with Pseudomonas aeruginosa is the main proven perpetrator of lung function decline and ultimate mortality in cystic fibrosis (CF) patients. Mucoid strains of this bacterium elaborate mucoid exopolysaccharide, also referred to as alginate. Alginate-based immunization of naïve animals elicits opsonic antibodies and leads to clearance of mucoid P. aeruginosa from the lungs. Alginate was isolated from mucoid P. aeruginosa strain 8821M by repeated ethanol precipitation, dialysis, proteinase and nuclease digestion, and chromatography. To improve immunogenicity, the purified antigen was coupled to tetanus toxoid (TT) with adipic acid dihydrazide (ADH) as a spacer and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDAC) as a linker. The reaction mixture was passed through a Sepharose CL-4B column. The resulting conjugate was composed of TT and large-size alginate polymer at a ratio of about 3 : 1; it was non-toxic and non-pyrogenic, and elicited high titres of alginate-specific IgG. Antisera raised against the conjugate had high opsonic activity against the vaccine strain. The alginate conjugate was also able to protect mice against a lethal dose of mucoid P. aeruginosa. These data indicate that an alginate-based vaccine has significant potential to protect against chronic infection with mucoid P. aeruginosa in the CF host.
Insights
A novel alginate conjugate vaccine shows promise for cystic fibrosis (CF) patients. This vaccine effectively clears Pseudomonas aeruginosa infections, reducing lung function decline and mortality in CF.
Area of Science:
- Bacteriology
- Immunology
- Vaccine Development
Background:
- Chronic Pseudomonas aeruginosa infection significantly impairs lung function and survival in cystic fibrosis (CF) patients.
- Mucoid strains of P. aeruginosa produce alginate, a key component contributing to chronic infections.
- Alginate-based immunization has demonstrated potential for clearing mucoid P. aeruginosa.
Purpose of the Study:
- To develop and evaluate an immunogenic and protective alginate-based vaccine against mucoid Pseudomonas aeruginosa.
- To improve the immunogenicity of purified alginate by conjugating it to tetanus toxoid (TT).
Main Methods:
- Alginate was isolated from mucoid P. aeruginosa strain 8821M using ethanol precipitation, dialysis, and enzymatic digestion.
- Purified alginate was conjugated to tetanus toxoid (TT) using adipic acid dihydrazide (ADH) and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDAC).
- The resulting alginate-TT conjugate was purified and characterized, and its immunogenicity and protective efficacy were tested in animal models.
Main Results:
- The alginate-TT conjugate was non-toxic and non-pyrogenic, eliciting high titers of alginate-specific IgG antibodies.
- Antisera generated against the conjugate exhibited significant opsonic activity against mucoid P. aeruginosa.
- The alginate conjugate vaccine protected mice against a lethal challenge with mucoid P. aeruginosa.
Conclusions:
- Alginate-based immunization is a viable strategy for combating chronic Pseudomonas aeruginosa infections in CF.
- The developed alginate-TT conjugate vaccine demonstrates significant potential for protecting CF patients against mucoid P. aeruginosa.
- This vaccine approach may reduce lung function decline and mortality associated with P. aeruginosa in cystic fibrosis.
