Immunochemical studies of the lipopolysaccharides of Hafnia alvei PCM 1219 and other strains with the O-antigens containing D-glucose 1-phosphate and 2-deoxy-2-[(R)-3-hydroxybutyramido]-D-glucose.
BACKGROUND
Hafnia alveiis the only species of the genus Hafnia, which belongs to the family of Enterobacteriaceae. These Gram-negative bacteria are commonly distributed in the natural environment and are often the cause of human opportunistic infections. Their lipopolysaccharides (LPSs) are important surface antigens which are responsible for the serological specificity and numerous cross-reactions with other enterobacterial genera. So far, 29 different O-polysaccharide (OPS, O-antigen) structures in Hafnias LPSs have been established and for some of them the molecular basis of the serological activity has been elucidated.


METHODS
OPS from H. alvei strain PCM 1219 was obtained by mild acid hydrolysis of the LPS followed by gel permeation chromatography of carbohydrate material on Sephadex G-50 column. The polysaccharide structure was determined using chemical methods as well as (13)C NMR and (1)H NMR spectroscopy. For serological studies, SDS-PAGE, immunoblotting, and passive hemagglutination tests were used.


RESULTS
The serological studies revealed a cross-reactivity of the LPSs of H. alvei PCM 1219 and a group of H. alvei strains with an O-antigen containing D-glucose 1-phosphate and [(R)-3-hydroxybutyramido]-D-glucose. The following structure of the OPS was established: where Acyl stands for (R)-3-hydroxybutyryl and the degree of O-acetylation is ~70%. The structure of the core oligosaccharide was found to be typical of the genus Hafnia.


CONCLUSIONS
Based on the OPS structure and serological results it was concluded that H. alvei strain PCM 1219 should be classified in the same serogroup as the H. alvei type strain ATCC 13337 and five other strains containing D-glucose 1-phosphate and 2-deoxy-2-[(R)-3-hydroxybutyramido]-D-glucose in their O-antigens.
Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114, Wrocław, Poland. katzenel@iitd.pan.wroc.pl
Ewa
E
Katzenellenbogen
Nina A
NA
Kocharova
Agnieszka
A
Korzeniowska-Kowal
Andrzej
A
Gamian
Maria
M
Bogulska
Bernadeta
B
Szostko
Alexander S
AS
Shashkov
Yuriy A
YA
Knirel
D000942:Antigens, Bacterial / Q000032:analysis / Q000276:immunology*; D005947:Glucose / Q000031:analogs & derivatives* / Q000737:chemistry; D005958:Glucosephosphates / Q000737:chemistry*; D006031:Glycosylation; D020611:Hafnia alvei / Q000145:classification / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D019081:O Antigens / Q000737:chemistry / Q000276:immunology*; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000276:immunology*; D013237:Stereoisomerism
C474835:2-deoxy-2-(3-hydroxybutyramido)-glucose; D000942:Antigens, Bacterial; D005958:Glucosephosphates; D019081:O Antigens; D011135:Polysaccharides, Bacterial; C031590:glucose-1-phosphate; D005947:Glucose