With a new flow cytometric cytotoxicity assay, we examined the mechanism of action of chimeric mouse human anti-CD20 monoclonal antibody IDEC-C2B8. IDEC-C2B8 alone induced direct cytotoxicity in four of eight examined CD20-expressing lymphoma cell lines (RAJI, DAUDI, JOK-1, and WT100) at a concentration above 100 ng/ml. Moreover, after 4 h incubation in human serum, only a moderate complement-dependent cellular cytotoxicity (CDCC) was observed, whereas cytotoxicity increased markedly after 3 days of culture, indicating that combined direct cytotoxicity and CDCC were responsible. IDEC-C2B8 induced an effective antibody-dependent cellular cytotoxicity (ADCC) in seven of eight tested lymphoma cell lines when peripheral blood mononuclear cells were used as effector cells. ADCC was moderately enhanced by cytokine interleukin-2, whereas interleukin-12, interferon-alpha, and GM-CSF had no influence. Interestingly, we could demonstrate a correlation between CD32 expression on lymphoma cell lines and IDEC-C2B8-induced direct cytotoxicity, indicating that crosslinking of CD20 with CD32 may be involved in the mechanism of cytotoxicity. We propose that direct cytotoxicity, CDCC, and ADCC result in the marked elimination of CD20-expressing ...Continue Reading
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