Anti-CD6-blocked ricin: an anti-pan T-cell immunotoxin.
We report the development of a potent anti-pan T-cell immunotoxin capable of killing cells in an antigen dependent manner. The immunotoxin is composed of a high affinity anti-CD6 antibody (IgG2a, Kd = 1.3 x 10(-11) M) conjugated to blocked ricin that is a chemically modified ricin molecule wherein the lectin binding sites of the B-chain have been blocked by covalent attachment of affinity ligands. Conjugation of blocked ricin to the antibody has minimal effect on the apparent affinity of the antibody and no effect on the ribosome-inactivating activity of the ricin A-chain moiety. Anti-CD6-blocked ricin is a specific and highly toxic immunoconjugate killing the antigen-positive Molt-4 cell line with an IC37 of 4 x 10(-12) M after a 24 h exposure of cells to the immunotoxin. Nonspecific cytotoxicity of anti-CD6-blocked ricin for the antigen-negative Namalwa cell line was more than 750-fold lower with an IC37 > 3 x 10(-9) M. The cytotoxicity of anti-CD6-blocked ricin is dependent on the length of the incubation of cells with the conjugate ranging from an IC37 of 1.5 x 10(-11) M leaving a surviving fraction of Molt-4 cells of 0.03 after a 2.5 h exposure to an IC37 of 5 x 10(-13) M and leaving a surviving fraction of 3 x 10(-6) after a continuous (3 weeks) exposure. Anti-CD6-blocked ricin is also capable of killing CD6 positive cells in human peripheral blood lymphocyte populations. Systemic toxicity of anti-CD6-blocked ricin in mice is similar to the toxicity of other immunotoxins containing blocked ricin that were found to be tolerated well by patients. An application of this immunoconjugate for the prevention and treatment of graft versus host disease or tissue graft rejection is suggested.
Collinson
A R
AR
ImmunoGen Inc., Cambridge, MA 02139.
Lambert
J M
JM
Liu
Y
Y
O'Dea
C
C
Shah
S A
SA
Rasmussen
R A
RA
Goldmacher
V S
VS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology; D000937:Antigen-Antibody Reactions; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D006801:Humans; D000922:Immunotoxins / Q000494:pharmacology* / Q000633:toxicity; D066298:In Vitro Techniques; D007700:Kinetics; D007928:Lethal Dose 50; D051379:Mice; D012276:Ricin / Q000037:antagonists & inhibitors / Q000494:pharmacology* / Q000633:toxicity; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; C062279:CD6 antigen; D000922:Immunotoxins; D012276:Ricin