Inhibition of a complement regulator in vivo enhances antibody therapy in a model of mammary adenocarcinoma

Membrane-bound complement regulatory proteins provide tumor cells with protection from antibody and complement in vitro. However, complement regulators are widely expressed on normal tissue, and inhibiting the function of complement regulatory proteins on tumor cells in vivo has not been investigated due to the absence of appropriate tumor-targeting strategies. Using a mouse model of rat mammary adenocarcinoma, we demonstrate that tumor-specific targeting of a complement regulator with a blocking antibody has functional consequences with regard to both complement deposition on tumor cells and the efficacy of monoclonal antibody therapy. Rat adenocarcinoma 13762 cells express Crry, a widely expressed rodent regulator of complement activation, and are recognized by C595 MAb, an anti-MUC1 MAb in clinical trials. Anti-rat Crry 5I2 MAb and F(ab)2 enhanced complement deposition on C595 MAb-sensitized 13762 cells in vitro. In vivo, C595 MAb bound to 13762 tumors, albeit not specifically, but was not therapeutic when administered after tumor challenge. However, the coadministration of 5I2 MAb with C595 MAb resulted in enhanced complement deposition and significantly delayed tumor onset and reduced tumor growth; 5I2 MAb alone also enhanced complement deposition and reduced tumor growth but less effectively than when combined with C595 MAb; 5I2 MAb alone did not directly activate mouse complement, but its inhibitory effect on Crry enhanced complement deposition following complement activation by both the alternative pathway and by natural IgM reactive to 13762 cells present in mouse serum. Our proof of principle study shows that inhibiting the function of a tumor-expressed complement regulatory protein enhances immune-mediated clearance of tumor cells and improves prospects for successful immunotherapy. The results justify further research and development of targeting strategies to inhibit or downregulate complement regulatory proteins on tumor cells. © 2004 Wiley-Liss, Inc.