The immunochemistry of antibodies sharing concanavalin A's anti-mannosyl binding specificity.
Procedures were developed for the synthesis of the disaccharide hapten, p-isothiocyanatophenyl 2-O-alpha-D-mannopyranosyl-alpha-D-mannopyranoside, and for its conjugation to hemocyanin. The synthetic carbohydrate: protein antigen was then emulsified in complete Freund's adjuvant and injected into the footpads of New Zealand White rabbits. A population of the resulting anti-conjugate antibodies displayed some binding properties analogous to concanavalin A, the carbohydrate-binding protein of the jack bean. The antisera weakly percipitated mannans from Saccharomyces rouxii, S. cerevisiae, and an alpha-(1 leads to 3)-mannopyranosyl transferase-deficient mutant from Kluyveromyces lactis Y58a. These polysaccharides, possessing side chains containing terminal alpha-(1 leads to 2)-mannobiosyl residues, produce strong percipitation reactions with concanavalin A. In addition, various saccharides were tested for their ability to inhibit the interaction of anti-conjugate antisera with alpha-(1 leads to 2)-mannobiosyl-containing polymers. p-Nitrophenyl 2-O-alpha-D-mannopyranosyl-alpha-D-mannopyranoside showed a strong complementarity for the binding sites of both the anti-conjugate antisera and concanavalin A. However, the antibody failed to bind a concanavalin A-reactive mouse fibrosarcoma or to stimulate mitogenesis of human peripheral lymphocytes.
D000818:Animals; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D003208:Concanavalin A / Q000494:pharmacology*; D004187:Disaccharides / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006241:Haptens; D007106:Immune Sera; D008213:Lymphocyte Activation; D008351:Mannans / Q000276:immunology; D008358:Mannose / Q000276:immunology*; D011234:Precipitins; D011817:Rabbits
D004187:Disaccharides; D006241:Haptens; D007106:Immune Sera; D008351:Mannans; D011234:Precipitins; D003208:Concanavalin A; D008358:Mannose