Cholera toxin as a mucosal adjuvant. Glutaraldehyde treatment dissociates adjuvanticity from toxicity.
Cholera toxin (CT), either mixed with or conjugated to unrelated protein Ag, is known to enhance the intestinal IgA response of rodents toward the unrelated Ag. Although relatively low doses of CT exert this gut mucosal adjuvant effect, the inherent toxicity of CT is a hindrance to its use in humans. Our report demonstrates that CT treated with 20 mM glutaraldehyde retains adjuvant properties but exhibits more than 1000-fold lower toxicity than untreated toxin. Glutaraldehyde was also used in a one-stage conjugation procedure to couple CT covalently to Sendai virus. Again, toxicity was reduced more than 1000-fold. This drop in toxicity is consistent with an observed 100-fold loss in binding capacity of the CT B subunit and a 20- to 50-fold reduction in adenylate cyclase activation by the CT A subunit. Oral administration of this virus-toxoid conjugate resulted in increased gut antiviral IgA titers compared with oral administration of either virus alone or of virus mixed with glutaraldehyde-treated toxin. This marked decrease in toxicity may afford a practical approach for the use of CT as a mucosal adjuvant.
Liang
X P
XP
Institute of Pathology, Case Western Reserve University, Cleveland, OH 44106.
Lamm
M E
ME
Nedrud
J G
JG
D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D000447:Aldehydes / Q000494:pharmacology; D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects; D002772:Cholera Toxin / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D005677:G(M1) Ganglioside; D005976:Glutaral / Q000494:pharmacology; D006028:Glycosphingolipids / Q000378:metabolism; D007070:Immunoglobulin A / Q000096:biosynthesis; D007413:Intestinal Mucosa / Q000276:immunology* / Q000378:metabolism; D008297:Male; D051379:Mice; D010222:Parainfluenza Virus 1, Human / Q000187:drug effects; D011817:Rabbits; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000032:analysis; D014121:Toxoids / Q000378:metabolism / Q000494:pharmacology / Q000633:toxicity; D014775:Virus Activation / Q000187:drug effects
D000276:Adjuvants, Immunologic; D000447:Aldehydes; D006028:Glycosphingolipids; D007070:Immunoglobulin A; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D014121:Toxoids; C034993:ganglioside receptor; D005677:G(M1) Ganglioside; C018835:asialo GM1 ganglioside; D002772:Cholera Toxin; D005976:Glutaral