Preparation of a diphtheria toxin-pullulan conjugate that elicits good IgG antibody production with poor IgE synthesis.
Diphtheria toxin is detoxified through conjugation with pullulan. The toxin-pullulan conjugate is easily purified by DEAE-Sephacel chromatography. The conjugate forms a transparent 'clear line' with anti-toxin antibodies on agarose plate, which offers a good indicator of conjugate formation. The toxin-pullulan conjugate induces both IgG1 and IgG2b antibody production with diminished IgE response, while the alum-precipitated conventional toxoid causes mainly increases in IgE as well as IgG1 antibody formation. The anti-toxin HA titre (IgG antibody) induced by the toxin-pullulan conjugate parallels the neutralizing activity of the immune-sera. These results suggest that the conjugation of toxin to pullulan is a very powerful method by which to develop a vaccine that induces neutralizing antibody with diminished IgE antibody synthesis.
Yamaya
S
S
Department of Applied Immunology, National Institute of Health, Shinagawa-ku, Tokyo.
Yamamoto
A
A
Komiya
T
T
Mizuguchi
J
J
Matuhasi
T
T
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D002850:Chromatography, Gel; D004167:Diphtheria Toxin / Q000276:immunology*; D004168:Diphtheria Toxoid / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005936:Glucans; D006386:Hemagglutination Tests; D005779:Immunodiffusion; D007073:Immunoglobulin E / Q000096:biosynthesis*; D007074:Immunoglobulin G / Q000096:biosynthesis*; D007075:Immunoglobulin M / Q000096:biosynthesis; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009500:Neutralization Tests; D051381:Rats; D011919:Rats, Inbred Strains
D000907:Antibodies, Bacterial; D004167:Diphtheria Toxin; D004168:Diphtheria Toxoid; D005936:Glucans; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007073:Immunoglobulin E; C009109:pullulan