Serological characterization of anti-endotoxin serum directed against the conjugate of oligosaccharide core ofEscherichia colitype R4 with tetanus toxoid

Abstract

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.

Epitope of the Vaccine-TypeBordetella pertussisStrain 186 Lipooligosaccharide and Antiendotoxin Activity of Antibodies Directed against the Terminal Pentasaccharide-Tetanus Toxoid Conjugate

“…The antibodies produced against complete core structures of Escherichia coli reacted strongly with LPS in the presence of serum proteins and were able to recognize LPS present on the surface of live bacteria (25,26,28).…”

Interaction of Mannose-Binding Lectin With Lipopolysaccharide Outer Core Region and Its Biological Consequences

“…H. alvei LPS is also an example of endotoxin having the E. coli -type structure of lipid A ( 22 – 24 ). A few strains of H. alvei synthesize LPS containing E. coli R4 [strains Polish Collection of Microorganisms (PCM) 23 or 1222] or Salmonella Ra (strain PCM 1212) core types (Figure 1 ) ( 25 , 26 ). The OS1 hexasaccharide is the predominant core OS for this species, with Hep and Kdo residues in its inner core region like most Gram-negative bacteria (Table 1 , footnote f) ( 24 , 27 ).…”

Lactoferrin-monophosphoryl lipid A complex enhances immunity of mice to Plesiomonas shigelloides CNCTC 138/92.

“…Sera were isolated and kept frozen at -20°C until determination of antibodies. Determination of immunoglobulin concentrations was performed according to a modified method described by (Lukasiewicz et al, 2003). Briefly, 96-well plates (Nunc) were coated using a 10 μg/ml solution of P. shigelloides LPS in 0.05 M sodium carbonate/bicar-bonate buffer (pH 9.6) at room temperature for 2 h and further 16 h at 4°C.…”