Isatuximab (Isa) is an IgG1 monoclonal antibody (Ab) that specifically recognizes human CD38. Once Isa engages multiple myeloma (MM) cells expressing a high level of CD38, it can induce tumor cell killing via Fc-dependent mechanisms including Ab-dependent cell-mediated cytotoxicity (ADCC), Ab-dependent cellular phagocytosis (ADCP), and complement-dependent cytotoxicity (CDC). To better understand the mechanism of Isa-mediated cytotoxicity, we studied CD38 levels in 16 established MM lines and measured Isa-mediated ADCC, ADCP, and CDC in tumor cell killing. The cytotoxic functions of Isa were dependent on CD38 receptor density (RD) in most cell lines. Isa-mediated ADCC was observed in a subset (7/16) of MM cell lines that displayed CD38 RD >100,000 molecules/cell. Similarly, the same subset of MM cells, with 1 exception, were sensitive to Isa-mediated ADCP, indicating a similar CD38 RD is also required for this killing effect. A much higher CD38 RD (>250,000 molecules/cell) alone was insufficient for Isa-mediated CDC. Cell lines MOLP-8 and MOLP-2 have CD38 RD >250,000 molecules/cell, but are resistant to Isa-mediated CDC. Overexpression of CD38 in cell lines with low endogenous CD38 expression did not always sensitize the cells to Isa-mediated CDC. These results suggest that additional mechanisms are involved in the regulation of such cytotoxic effects. We further investigated the expression of complement-cascade inhibitors CD46, CD55, and CD59. High-level expression of at least one of these molecules was associated with resistance to Isa-mediated CDC, even when cells were CD38 high (hi). By comparing 4 selected MM cell lines, we found that a minimal RD level of 50,000 molecules/cell for CD46, CD55 or CD59 appears to be an important threshold to suppress Isa-mediated CDC. When CD59 function was neutralized by an anti-CD59 antibody, we were able to re-sensitize the cells to Isa-mediated CDC in CD38hi MOLP-8 cells. Neutralizing CD59 function alone did not rescue Isa-mediated CDC in CD38-low expressing NCI-H929 cells. However, overexpressing CD38 and inhibiting CD59 rendered NCI-H929 cells sensitive to CDC. Taken together, the high-level of CD38 expression and low-level of CD59 (and perhaps other inhibitors) expression are important for Isa-mediated CDC in killing of target tumor cells. In conclusion, the main immune effector mechanisms involved in Isa-mediated killing of MM cells include ADCC and ADCP. These effects are dependent on high levels of CD38 RD in MM cell lines in vitro. Further confirmation is under way using samples from MM patients.

Citation Format: Zhili Song, Guang Yang, Anlai Wang, Rita Greco, Joachim Theilhaber, Elvis Shehu, Daniel Ajona, Bruno Paiva, Chen Zhu, Dmitri Wiederschain, Marielle F. Chiron Blondel, Francisco Adrian. Isatuximab-induced multiple myeloma cell killing through effector functions is dependent on CD38 expression and complement inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2966.