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. 1995 Mar;40(2):88–94. doi: 10.1007/BF01520289

Increased tumor cell reactivity and complement-dependent cytotoxicity with mixtures of monoclonal antibodies against different gangliosides

Shengle Zhang 1,✉, Friedhelm Helling 1, Kenneth O Lloyd 1, Philip O Livingston 1
PMCID: PMC11037804  PMID: 7882387

Abstract

Melanomas and other cancers of neuroectodermal origin express multiple cell-surface gangliosides in patterns that vary significantly even within the same tumor type. Monoclonal antibodies (mAb) against four of these gangliosides (GM2, GD2, 9-O-acetyl-GD3 and GD3) were tested alone and in combination on 14 tumor cell lines (7 melanomas, 3 neuroblastomas, 3 sarcomas and 1 astrocytoma) using flow cytometry and complement-dependent cytotoxicity (CDC) assays. Increased tumor cell recognition and CDC resulting from the combination of three or four mAb were found in 14/14 tested cell lines, and this was most striking when each mAb was used at suboptimal concentration. At these concentrations, the average mean fluorescence intensity of the 14 cell lines with individual mAb was between 3.0 and 6.8 and increased to 10.8 and 18.8 with the three- and four-mAb mixtures. The average percentage CDC-specific release with individual mAb was 2.0%–8.3%, and 12.3% and 16.6% with the three- and four-mAb combinations. The number of cell lines showing significant mean fluorescence intensity and CDC increased from 2–8/14 with single mAb to 13–14/14 with the mixtures of three or four mAb. Our experimental results support the rationale for active immunization with a polyvalent ganglioside vaccine or passive therapy with a combination of mAb to different gangliosides in patients with tumors of neuroectodermal origin. In addition, our studies have demonstrated that 9-O-acetyl-GD3 is a surprisingly effective target for immune attack, although it is a minor constituent of these cells.

Key words: Monoclonal antibody, Ganglioside Neuroectodermal tumor, Immunotherapy, Polyvalent vaccine

References

  • 1.Berd D, Herlyn M, Koprowski H, Mastrangelo MJ. Flow cytometric determination of the frequency and heterogeneity of expression of human melanoma-associated antigens. Cancer Res. 1989;49:6840. [PubMed] [Google Scholar]
  • 2.Blomberg K, Granberg C, Hemmila I, Lovgren T. Europium-labelled target cells in an assay of natural killer cell activity. I. A novel non-radioactive method based on time-resolved fluorescence. J Immunol Methods. 1986;86:225. doi: 10.1016/0022-1759(86)90457-6. [DOI] [PubMed] [Google Scholar]
  • 3.Cheresh DA, Varki AP, Varki NM, Stallcup WB, Levine JM, Reisfeld RA. A monoclonal antibody recognizes anO-acetylated sialic acid in a human melanoma-associated ganglioside. J Biol Chem. 1984;259:7453. [PubMed] [Google Scholar]
  • 4.Cheung N-K V, Saarinen UM, Neely JE, Landmeier B, Donovan D, Coccia PF. Monoclonal antibodies to a glycolipid antigen on human neuroblastoma cells. Cancer Res. 1985;45:2642. [PubMed] [Google Scholar]
  • 5.Cheung N-K V, Lazarus H, Miraldi FD, Abramowsaky CR, Kallic S, Saarinen UM, Spitzer T, Strandjord SE, Coccia PF, Berger NA. Ganglioside GD2 specific monoclonal antibody 3F8: a phase-I study in patients with neuroblastoma and malignant melanoma. J Clin Oncol. 1987;5:1430. doi: 10.1200/JCO.1987.5.9.1430. [DOI] [PubMed] [Google Scholar]
  • 6.Cui J, Bystryn J-C. An improved europium release assay for complement-mediated cytolysis. J Immunol Methods. 1992;147:13. doi: 10.1016/s0022-1759(12)80023-8. [DOI] [PubMed] [Google Scholar]
  • 7.Dippold WG, Lloyd KO, Li LT, Ikeda H, Oettgen HF, Old LJ. Cell-surface antigens of human malignant melanoma: definition of six antigenic systems with monoclonal antibodies. Proc Natl Acad Sci USA. 1980;77:6114. doi: 10.1073/pnas.77.10.6114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Hamilton WB, Helling F, Lloyd KO, Livingston PO. Ganglioside expression on human malignant melanoma assessed by quantitative immune thin-layer chromatography. Int J Cancer. 1993;53:566. doi: 10.1002/ijc.2910530407. [DOI] [PubMed] [Google Scholar]
  • 9.Hamilton WB, Helling F, Livingston PO. Ganglioside expression on sarcoma and small-cell lung carcinoma compared to tumors of neuroectodermal origin (abstract) Proc Am Assoc Cancer Res. 1993;34:491. [Google Scholar]
  • 10.Houghton AN, Mintzer D, Cordon-Cardo C, Welt S, Fliegel B, Vadhan S, Carswell E, Melamed MR, Oettgen HF, Old LJ. Mouse monoclonal IgG3 antibody detecting GD3 ganglioside: a phase-I trial in patients with malignant melanoma. Proc Natl Acad Sci USA. 1985;82:1242. doi: 10.1073/pnas.82.4.1242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Irie RF, Morton DL. Regression of cutaneous metastatic melanoma by introlesional injection with human monoclonal antibody to ganglioside GD2. Proc Natl Acad Sci USA. 1986;83:8694. doi: 10.1073/pnas.83.22.8694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Irie RF, Matsuki T, Morton DL. Human monoclonal antibody to ganglioside GM2 for melamoma treatment. Lancet. 1989;1:786. doi: 10.1016/s0140-6736(89)92606-8. [DOI] [PubMed] [Google Scholar]
  • 13.Kanda S, Cochran AJ, Lee WR, Morton DL, Irie RF. Variations in the ganglioside profile of uveal melanoma correlate with cytologic heterogeneity. Int J Cancer. 1992;52:682. doi: 10.1002/ijc.2910520503. [DOI] [PubMed] [Google Scholar]
  • 14.Livingston PO, Ritter F, Srivastava P, Padavan M, Calves MJ, Oettgen HF, Old LJ. Characterization of IgG and IgM antibodies induced in melanoma patients by immunization with purified GM2 ganglioside. Cancer Res. 1989;49:7045. [PubMed] [Google Scholar]
  • 15.Livingston PO, Wong GYC, Adluri S, Tao Y, Padavan M, Parente R, Hanlon C, Calves MJ, Helling F, Ritter G, Oettgen HF, Old LJ. Improved survival in stage III melanoma patients with GM2 antibodies: a randomized trial of adjuvant vaccination with GM2 ganglioside. J Clin Oncol. 1994;12:1036. doi: 10.1200/JCO.1994.12.5.1036. [DOI] [PubMed] [Google Scholar]
  • 16.Lloyd KO, Gordon CM, Thampoe IJ, DiBenedetto C. Cell surface accessibility of individual gangliosides in malignant melanoma cells to antibodies is influenced by the total ganglioside composition of the cells. Cancer Res. 1992;52:4948. [PubMed] [Google Scholar]
  • 17.Natoli EJ, Jr, Livingston PO, Pukel CS, Lloyd KO, Wiegandt H, Szalay J, Oettgen HF, Old LJ. A murine monoclonal antibody detectingN-acetyl andN-glycoly GM2: characterization of cell surface reactivity. Cancer Res. 1986;46:4116. [PubMed] [Google Scholar]
  • 18.Ravindranath MH, Morton DL, Irie RF. An epitope common to gangliosidesO-acetyl-GD3 and GD3 recognized by antibodies in melanoma patients after active specific immunotherapy. Cancer Res. 1989;49:3891. [PubMed] [Google Scholar]
  • 19.Ritter G, Boosfeld E, Adluri R, Calves M, Oettgen HF, Old LJ, Livingston PO. Antibody response to immunization with ganglioside GD3 and GD3 congeners (lactones, amide and gangliosidol) in patients with malignant melanoma. Int J Cancer. 1991;48:379. doi: 10.1002/ijc.2910480312. [DOI] [PubMed] [Google Scholar]
  • 20.Saleh MN, Khazaeli MB, Wheeler RH, Dropcho E, Liu T, Urist M, Miller DM, Lawson S, Dixon P, Russell CH, LoBuglio AF. Phase I trial of the murine monoclonal anti-GD2 antibody 14G2a in metastatic melanoma. Cancer Res. 1992;52:4342. [PubMed] [Google Scholar]
  • 21.Svennerholm L. Chromatographic separation of human brain gangliosides. J Neurochem. 1963;10:613. doi: 10.1111/j.1471-4159.1963.tb08933.x. [DOI] [PubMed] [Google Scholar]
  • 22.Tai T, Cahan LD, Tsuchida T, Saxton RE, Irie RF, Morton DL. Immunogenicity of melanoma-associated gangliosides in cancer patients. Int J Cancer. 1985;35:607. doi: 10.1002/ijc.2910350507. [DOI] [PubMed] [Google Scholar]
  • 23.Tsuchida T, Ravindranath MH, Saxton RE, Irie RF. Gangliosides of human melanoma: altered expression in vivo and vitro. Cancer Res. 1987;47:1278. [PubMed] [Google Scholar]
  • 24.Tsuchida T, Saxton RE, Morton DL, Irie RF. Gangliosides of human melanoma. J Nat Cancer Inst. 1987;78:45. doi: 10.1093/jnci/78.1.45. [DOI] [PubMed] [Google Scholar]
  • 25.Tsuchida T, Saxton RE, Morton DL, Irie RF. Gangliosides of human melanoma. Cancer. 1989;63:1166. doi: 10.1002/1097-0142(19890315)63:6<1166::aid-cncr2820630621>3.0.co;2-5. [DOI] [PubMed] [Google Scholar]

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