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J Med Microbiol 55 (2006), 1441-1446; DOI: 10.1099/jmm.0.46696-0
© 2006 Society for General Microbiology
ISSN 1473-5644

Synthesis and characterization of Pseudomonas aeruginosa alginate–tetanus toxoid conjugate

Nasim Kashef1, Qorban Behzadian-Nejad1, Shahin Najar-Peerayeh1, Kamran Mousavi-Hosseini2, Mohammad Moazzeni3 and Gholamreza Esmaeeli Djavid4

1 ,3 Department of Bacteriology1 and Department of Immunology3 , School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Iranian Blood Transfusion Organization, Research Center, Tehran, Iran

4 Academic Center for Education, Culture, and Research, Tehran, Iran

Correspondence
Nasim Kashef
na_kashef{at}yahoo.com

Received 24 April 2006
Accepted 13 July 2006

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.

Abbreviations: ADH, adipic acid dihydrazide; CF, cystic fibrosis; EDAC, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide; i.p., intraperitoneal(ly); KLH, keyhole-limpet haemocyanin; MEP, mucoid exopolysaccharide; TT, tetanus toxoid.






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