The capacity of highly purified preparations of ricin A and B chains to reconstitute ricin toxicity both in vitro and in vivo was studied. When the nontoxic A and B chain subunits were mixed and electrophoresed on sodium dodecyl sulphate-polyacrylamide gels (SDS-PAGE), reconstituted ricin was observed. The mixtures killed cells of the human Burkityt's lymphoma cell line Daudi in vitro and killed mice after i.v. injection. It was also shown that when mice were injected with one ricin subunit followed by administration of the complementary polypeptide up to 8 hr later, they died with lesions similar to those of ricin-induced death. This observation suggests that A and B chains recombine either in the serum or on the surface of cells. The rate of clearance of A and B chains from the blood of rats indicated that sufficient concentrations of either chain were present in the circulation 8 hr after injection to account for the observed toxicity. The above studies therefore suggest that the subunits of ricin have a very high affinity for each other and are capable of reconstituting biologically active ricin in vitro and in injected mice.
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