Characterization of the increased cytotoxicity of gelonin anti-T cell immunoconjugates compared with ricin A chain immunoconjugates.
Ribosomal inactivating proteins such as gelonin (Gel) and ricin A chain (RTA) conjugated to MoAbs bind to specific target cells, and upon internalization inhibit protein synthesis, ultimately resulting in cell death. We report here that Gel anti-T cell MoAb conjugates are more cytotoxic than RTA conjugates when tested against human peripheral blood mononuclear cells (PBMC). This increased cytotoxicity is observed whether Gel is conjugated to the anti-T cell MoAb or to an anti-mouse immunoglobulin Fab' fragment which then binds to the murine anti-human T cell MoAb. Gel conjugates are not only effective at lower concentrations, but also produce a greater extent of inhibition of cellular proliferation. Moreover, a 10 min exposure to a Gel conjugate is as effective as a 90 h exposure to an RTA conjugate. When part of anti-T cell F(ab')2 or Fab' conjugates, Gel affects the early steps in cellular intoxication more than RTA; Gel conjugates bind more avidly and accelerate the modulation of antigen. In contrast, when part of whole IgG conjugates, Gel does not affect the binding to or modulation of surface antigen compared with RTA, while it does increase conjugate cytotoxicity. These observations suggest that Gel may be delivered more efficiently into the cytosol than RTA. A divergent intracellular pathway for Gel is also supported by the inability of chemical potentiators, which strongly enhance RTA potency, to affect Gel potency. These properties of Gel might also be advantageous for immunoconjugates made with other MoAbs or receptor-binding molecules.
Fishwild
D M
DM
Department of Immunology, XOMA Corporation, Berkeley, CA.
Wu
H M
HM
Carroll
S F
SF
Bernhard
S L
SL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000961:Antilymphocyte Serum / Q000494:pharmacology; D001667:Binding, Competitive; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007167:Immunotherapy; D000922:Immunotoxins / Q000494:pharmacology*; D066298:In Vitro Techniques; D051379:Mice; D010940:Plant Proteins / Q000494:pharmacology*; D054789:Ribosome Inactivating Proteins, Type 1; D012276:Ricin / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000961:Antilymphocyte Serum; D000922:Immunotoxins; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; C025674:GEL protein, Gelonium multiflorum; D012276:Ricin