title: Neutralizing monoclonal antibodies against ricin's enzymatic subunit interfere with protein disulfide isomerase-mediated reduction of ricin holotoxin in vitro.
abstract: The penultimate event in the intoxication of mammalian cells by ricin toxin is the reduction, in the endoplasmic reticulum (ER), of the intermolecular disulfide bond that links ricin's enzymatic (RTA) and binding (RTB) subunits. In this report we adapted an in vitro protein disulfide isomerase (PDI)-mediated reduction assay to test the hypothesis that the RTA-specific neutralizing monoclonal antibody (mAb) IB2 interferes with the liberation of RTA from RTB. IB2 recognizes an epitope located near the interface between RTA and RTB and, like a number of other RTA-specific neutralizing mAbs, is proposed to neutralize ricin intracellularly. In this study, we found that IB2 virtually eliminated the reduction of ricin holotoxin into RTA and RTB in vitro. Surprisingly, three other neutralizing mAbs (GD12, R70 and SyH7) that bind epitopes at considerable distance from ricin's disulfide bond were as effective (or nearly as effective) as IB2 in interfering with PDI-mediated liberation of RTA from RTB. By contrast, two non-neutralizing RTA-specific mAbs, FGA12 and SB1, did not affect PDI-mediated reduction of ricin. These data reveal a possible mechanism by which RTA-specific antibodies may neutralize ricin intracellularly, provided they are capable of trafficking in association with ricin from the cell surface to the ER.
authors.affiliation: Division of Infectious Disease, Wadsworth Center, New York State Department of Health, Albany, NY 12208, United States.
authors.forename: Joanne M
authors.initials: JM
authors.lastname: O'Hara
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Mantis
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D057134:Antibodies, Neutralizing / Q000378:metabolism*; D000918:Antibody Specificity; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D008958:Models, Molecular; D010084:Oxidation-Reduction; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology; D012276:Ricin / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000276:immunology*; D014118:Toxins, Biological / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology; D014709:Vero Cells
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D007074:Immunoglobulin G; D021122:Protein Subunits; D014118:Toxins, Biological; D012276:Ricin; D019704:Protein Disulfide-Isomerases
keywords: Antibody; Intracellular; Neutralization; Toxin