Characterization of a Proapoptotic Antiganglioside GM2 Monoclonal Antibody and Evaluation of Its Therapeutic Effect on Melanoma and Small Cell Lung Carcinoma Xenografts

Abstract

Monoclonal antibodies have begun to show great clinical promise for the treatment of cancer. Antibodies that can directly affect a tumor cell's growth and/or survival are of particular interest for immunotherapy. Previously, we described monoclonal antibody DMF10.62.3 that had antiproliferative and proapoptotic effects when it bound an antigen of unknown identity on tumor cells in vitro. In this report, we determined that DMF10.62.3 and a clonally related antibody DMF10.167.4 recognize the ganglioside GM2. These antibodies react with a GM2 epitope that is expressed on a large number of tumor cell lines, including human melanoma and small cell lung carcinoma, but not on normal primary lines or most normal tissues. Interestingly, this pattern of cellular reactivity is distinct from that reported for other previously described GM2 antibodies, a difference that is presumably due to DMF10.167.4's binding to a unique GM2-associated epitope. Additional characterization of DMF10.167.4 revealed that this antibody was able to induce apoptosis and/or block cellular proliferation when cultured in vitro with the human Jurkat T lymphoma, CHL-1 melanoma, and SBC-3 small cell lung carcinoma lines. In vivo , DMF10.167.4 antibody was well tolerated in mice and did not detectably bind to or damage normal tissues. However, this antibody was able to prevent murine E710.2.3 lymphoma, human CHL-1 melanoma, and SBC-3 small cell lung carcinoma lines from establishing tumors in vivo and blocked progression of established CHL-1 and SBC-3 tumors in vivo. Therefore, monoclonal antibody DMF10.167.4 has immunotherapeutic potential. (Cancer Res 2005; 65(14): 6425-34)

Anti-ganglioside antibody-induced tumor cell death by loss of membrane integrity

“…However, the possible activation of these signaling processes by anti-ganglioside mAbs is still unknown. There are only evidences of apoptosis induction in melanoma and small cell lung cancer cells by anti-GM2 and anti-GD2 antibodies, respectively (15,16). In the present report, we described for the first time that an anti-NGcGM3 antibody induced a novel cell death mechanism.…”

TNF-α Induction of GM2 Expression on Renal Cell Carcinomas Promotes T Cell Dysfunction

“…Indeed, mAbs with specificity for GM2 bind epitopes on a large number of tumor lines, but do not recognize normal primary lines or most normal tissues (24). These Abs have been shown to mediate both complement-dependent cytotoxicity and Ab-dependent cellular cytotoxicity (53), which variously block the proliferation of some tumor cells in vitro, inhibit the outgrowth of micrometastases (54), and/or prevent the establishment of murine tumors in vivo (24,55,56).…”

“…Indeed, mAbs with specificity for GM2 bind epitopes on a large number of tumor lines, but do not recognize normal primary lines or most normal tissues (24). These Abs have been shown to mediate both complement-dependent cytotoxicity and Ab-dependent cellular cytotoxicity (53), which variously block the proliferation of some tumor cells in vitro, inhibit the outgrowth of micrometastases (54), and/or prevent the establishment of murine tumors in vivo (24,55,56). Mixed results, however, have been reported for related human trials, as follows: whereas anti-GM2 mAbs have been found to successfully eradicate melanoma metastases (57), in some cases anti-murine Ab responses are induced that interfere with and inhibit efficacy of the treatment (58).…”

“…To quantify the TNF-mediated change in SK-RC-45-associated gangliosides for which specific commercial Abs were available, ELISAs were set up, as described previously (24). Briefly, standard bovine brain-derived gangliosides and gangliosides derived from tumor cell lines or tumor tissue were resuspended in a 1:1 solution of CHCl 3 :MeOH at 1 mg/ml, and appropriate aliquots of these solutions were added to individual wells containing 100 l of MeOH.…”

GM2 Expression in Renal Cell Carcinoma: Potential Role in Tumor-Induced T-Cell Dysfunction

“…A hamster monoclonal anti-GM2 antibody (DMF10.167.4) was a gift from Corixa Corp. (Seattle, WA) and Dr. Kenneth Rock (Department of Pathology, University of Massachusetts Medical School, Worcester, MA; ref. 27). Ficoll-Hypaque was purchased from Amersham Pharmacia Biotech AB (Uppsala, Sweden).…”