Serological characterization of anti-endotoxin serum directed against the conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid

• Published in: FEMS immunology and medical microbiology Vol. 37; no. 1; pp. 59 - 67
• Main Authors: Lukasiewicz, Jolanta, Jachymek, Wojciech, Niedziela, Tomasz, Dzieciatkowska, Monika, Lakomska, Joanna, Miedzybrodzki, Ryszard, Fortuna, Wojciech, Szymaniec, Stanislaw, Misiuk-Hojlo, Marta, Lugowski, Czeslaw
• Format: Journal Article
• Language: English
• Published: Oxford, UK Elsevier B.V 10.06.2003; Blackwell Publishing Ltd; Blackwell
• Subjects: Animals; Antibody; Bacteriology; Biological and medical sciences; Carbohydrate Sequence; Cell Line; Core region; Endotoxin; Endotoxins - immunology; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Escherichia coli - chemistry; Escherichia coli - immunology; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Glycoconjugates - immunology; Immune Sera - immunology; Immunoblotting; Lipopolysaccharides - chemistry; Lipopolysaccharides - immunology; Lung - cytology; Lung - immunology; Macrophages - immunology; Mice; Microbiology; Molecular Sequence Data; Neoglycoconjugate; Nitric Oxide - biosynthesis; Nitric Oxide - immunology; Tetanus Toxoid - chemistry; Tetanus Toxoid - immunology; Tumor Necrosis Factor-alpha - biosynthesis; Tumor Necrosis Factor-alpha - immunology; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies; R4; Antibody; Endotoxin; Escherichia coli; Neoglycoconjugate; Core region; Enzyme; Oligosaccharide; Metalloendopeptidases; Peptidases; Toxin; Lipopolysaccharide; Bacteria; Hydrolases; Serum; Tentoxilysin; Enterobacteriaceae
• ISSN: 0928-8244; 1574-695X
• DOI: 10.1016/S0928-8244(03)00104-4

The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti-lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme-linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti-core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti-OS R4-TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin-induced tumor necrosis factor α and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells.