Toxicity of xanthine oxidoreductase to malignant B lymphocytes.
Reactive oxygen species (ROS) generated by xanthine oxidoreductase (XOR) were toxic to B lymphoma-derived Raji cells (positive for 8A monoclonal antibody, mAb). The sensitivity of these malignant cells to the hypoxanthine/XOR system was higher than that observed in peripheral human lymphocytes. The understanding of the mechanisms of cytotoxicity induced by XOR-produced ROS is essential in view of a possible clinical application. Cell death mostly had the feature of apoptosis and post-apoptotic necrosis and depended on the activity of XOR. Catalase, but not superoxide dismutase, protected cells from the toxicity of XOR, thus indicating that cell damage depended on the production of hydrogen peroxide. The toxicity of ROS was selectively targeted to malignant Raji cells by antibody-XOR conjugation, either directly, with an 8A-XOR conjugate, or indirectly, with an 8A mAb plus an anti-mouse IgG-XOR. Both direct and indirect immunotoxins induced apoptotic death to target cells in a dose-dependent manner. These conjugates showed no aspecific cytotoxicity in conditions very similar to the ex vivo treatment of cell suspension for bone marrow transplantation. Moreover, the prevalence of apoptotic death over necrosis may reduce the in vivo inflammatory response and its local and systemic consequences, thus becoming relevant in the construction of immunotoxins with therapeutic potential.
Battelli
M G
MG
Department of Experimental Pathology, University of Bologna, Italy. mariagiulia.battelli@unibo.it
Polito
L
L
Falà
F
F
Musiani
S
S
Tazzari
P L
PL
Stirpe
F
F
Bolognesi
A
A
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001402:B-Lymphocytes / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D045744:Cell Line, Tumor; D006801:Humans; D000922:Immunotoxins / Q000378:metabolism / Q000633:toxicity; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016393:Lymphoma, B-Cell / Q000201:enzymology / Q000473:pathology / Q000628:therapy; D051379:Mice; D009336:Necrosis; D017382:Reactive Oxygen Species / Q000378:metabolism / Q000633:toxicity; D014969:Xanthine Oxidase / Q000378:metabolism*
D000922:Immunotoxins; D017382:Reactive Oxygen Species; D007770:L-Lactate Dehydrogenase; D014969:Xanthine Oxidase