- Meeting abstract
- Open Access
A new type of glycoconjugate vaccine containing Klebsiellafimbriae type 1 and 3 as carrier proteins
© Current Science Ltd 1997
- Published: 1 March 1997
- Ammonium Sulfate
- Carrier Protein
- Conjugate Vaccine
- Ammonium Sulfate Precipitation
Opportunistic pathogens continue to be a major cause of infections among hospitalized patients. The vaccination as a primary defence strategy and the development of new vaccines of broad specificity offer an important tool to use against bacterial infections. Our study concentrates on a glycoconjugate vaccine based on covalently attached bacterial antigen to Klebsiella fimbriae type 1 and 3.
Fimbriae mediate the attachment of many pathogenic bacteria to host cells. The mannose-specific fimbriae of type 1 are expressed in many enterobacterial species. The mannose-resistant type 3 of fimbriae are produced by majority strains of Klebsiella . These surface antigens may serve as carrier proteins and also as a common antigen for vaccine of broad specificity. For the studies of stimulation of immune mediators it is necessary to have the fimbrial preparations free of endotoxin. In order to isolate pure fimbriae, simple methods have been adopted, especially considering the contamination with lipopolysaccharide. The procedure involves homogenization, ammonium sulfate precipitation, gel filtration in the presence of 6 M urea and hydrophobic chromatography on phenyl-sepharose. The purity was checked in SDS-PAGE, with silver staining specific for proteins and lipopolysaccharides as well as in mass spectrometry with MALDI-TOF technique, which allowed also to determine the precise molecular mass of fimbrial monomers. We have also undertaken the studv on stimulation of cytokine induction by fimbriae. The type 3 of Klebsiella fimbriae are moderate inductors of IL-6 and interferon, whereas type 1 is even less potent inductor. Both types of fimbriae are almost inactive regarding the stimulation of TNF when tested in human whole blood assay. These results prompt us for studies on practical use of fimbriae, especially type 1, as carriers for conjugate vaccine. Therefore we conjugated fimbrial protein with a core oligosaccharide fraction obtained from Escherichia coli K-12 lipopolysaccharide, which has been found to contain an epitope common for several enterobacterial species . The conjugate was immunogenie in rabbit model regarding to the oligosaccharide hapten and fimbrial carrier, respectively.
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