Affinity cytotoxicity with an alcohol dehydrogenase-antibody conjugate and allyl alcohol.
A potent new enzyme-antibody conjugate system for amplifying cytotoxicity was tested in a well-defined model of hapten [2,4,6-trinitrophenyl (TNP)]-substituted tumor cells (HEp2) and purified anti-hapten antibody. Brief treatment of TNP-HEp2 cells with low concentrations (0.05 to 0.74 micrograms/ml) of antihapten antibody-alcohol dehydrogenase conjugate (Ab-ADH) followed by culture in complement-free medium containing nicotinamide adenine dinucleotide and allyl alcohol or 2-fluoroethanol resulted in 15 to 90% cell killing as measured by 5-[125l]iodo'-2-deoxyuridine uptake assay. The importance of the complete enzyme system was indicated by reduced or absent cytotoxicity if Ab-ADH, nicotinamide adenine dinucleotide, or allyl alcohol (or 2-fluorethanol) were omitted. Immunological specificity of the Ab-ADH was demonstrated by reduced or absent cytotoxicity when: (a) HEp2 cells were not coated with TNP; (b) Ab-ADH binding onto TNP-cells was blocked by free hapten (2,4-dinitrophenyllysine); or (c) unconjugated alcohol dehydrogenase and anti-TNP purified IgG anti-2,4,6-trinitrophenyl antibody with NAD+ and allyl alcohol or anti-TNP antibody with complement were used.
D000433:1-Propanol / Q000276:immunology; D000429:Alcohol Oxidoreductases / Q000276:immunology*; D000498:Allyl Compounds / Q000276:immunology; D000912:Antibodies, Neoplasm; D000918:Antibody Specificity; D000920:Antibody-Dependent Cell Cytotoxicity; D002478:Cells, Cultured; D003165:Complement System Proteins; D006241:Haptens / Q000008:administration & dosage; D066298:In Vitro Techniques; D009243:NAD / Q000008:administration & dosage; D009374:Neoplasms, Experimental / Q000276:immunology; D020005:Propanols; D014301:Trinitrobenzenes / Q000276:immunology
D000498:Allyl Compounds; D000912:Antibodies, Neoplasm; D006241:Haptens; D020005:Propanols; D014301:Trinitrobenzenes; D009243:NAD; D003165:Complement System Proteins; D000433:1-Propanol; D000429:Alcohol Oxidoreductases