Serological characterization of anti-endotoxin serum directed against the conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid.
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 alpha and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells.
Lukasiewicz
Jolanta
J
Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland. czaja@iitd.pan.wroc.pl
Jachymek
Wojciech
W
Niedziela
Tomasz
T
Dzieciatkowska
Monika
M
Lakomska
Joanna
J
Miedzybrodzki
Ryszard
R
Fortuna
Wojciech
W
Szymaniec
Stanislaw
S
Misiuk-Hojlo
Marta
M
Lugowski
Czeslaw
C
D000818:Animals; D002240:Carbohydrate Sequence; D002460:Cell Line; D004731:Endotoxins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000737:chemistry / Q000276:immunology*; D005434:Flow Cytometry; D006001:Glycoconjugates / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D015151:Immunoblotting; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology*; D008168:Lung / Q000166:cytology / Q000276:immunology; D008264:Macrophages / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000096:biosynthesis / Q000276:immunology; D013745:Tetanus Toxoid / Q000737:chemistry / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000276:immunology
D004731:Endotoxins; D006001:Glycoconjugates; D007106:Immune Sera; D008070:Lipopolysaccharides; D013745:Tetanus Toxoid; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide