Measurement of the human immune response to meningococcal lipooligosaccharide antigens by using serum to inhibit monoclonal antibody binding to purified lipooligosaccharide.
We developed a human inhibition monoclonal enzyme-linked immunosorbent assay (HIMELISA) to investigate the human immune response to the lipooligosaccharides (LOS) of Neisseria meningitidis. Monoclonal antibodies (MAb) were used to define seven epitopes on four LOS molecules of a meningococcal strain (126E) previously shown to express immunogenic LOS epitopes. The assay could distinguish epitope-specific antibody within whole sera. Neither the specificity nor the amount of the antibody measured by HIMELISA in sera of vaccinates changed during the immune response to meningococcal capsular polysaccharides, a chemically unrelated antigen. By using the HIMELISA, it was determined that sera from adults convalescing from meningococcal disease strongly inhibited MAb binding to two of the seven defined epitopes. The 3.6-kilodalton LOS of strain 126E expressed both of these epitopes. In addition, one of the inhibited epitopes was also expressed on the 4.0-kilodalton LOS of strain 126E. The convalescent-phase sera inhibited MAb binding to these two epitopes when they were expressed on LOS of diverse meningococcal strains. An acute-phase serum blocked MAb to the two epitopes to a lesser degree than did a convalescent-phase serum from the same patient. Immunoblotting the sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated LOS with convalescent-phase sera confirmed the specificity of the human anti-LOS antibody identified by HIMELISA.
Centre for Immunochemistry, Veterans Administration Medical Center, San Francisco, California 94121.
M M
MM
Estabrook
R E
RE
Mandrell
M A
MA
Apicella
J M
JM
Griffiss
D000907:Antibodies, Bacterial / Q000032:analysis*; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D003289:Convalescence; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D015151:Immunoblotting; D008070:Lipopolysaccharides / Q000276:immunology*; D008589:Meningococcal Infections / Q000276:immunology; D009345:Neisseria meningitidis / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007106:Immune Sera; D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides