1. Affinity cytotoxicity with an alcohol dehydrogenase-antibody conjugate and allyl alcohol.
- Author
-
Philpott GW, Grass EH, and Parker CW
- Subjects
- 1-Propanol immunology, Allyl Compounds immunology, Antibody Specificity, Cells, Cultured, Complement System Proteins, Haptens administration & dosage, In Vitro Techniques, NAD administration & dosage, Neoplasms, Experimental immunology, Trinitrobenzenes immunology, Alcohol Oxidoreductases immunology, Antibodies, Neoplasm, Antibody-Dependent Cell Cytotoxicity, Propanols
- Abstract
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. more...
- Published
- 1979