Skip to main content
SpringerLink
Log in

Antibody dependent cellular phagocytosis (ADCP) and antibody dependent cellular cytotoxicity (ADCC) of breast cancer cells mediated by bispecific antibody, MDX-210

  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Background: MDX-210 is a bispecific antibody (BsAb) with specificity for both the proto-oncogene product of HER-2/neu (c-erbB-2) and FcγRI (CD64). HER-2/neu is overexpressed in malignant tissue of approximately 30% of patients with breast cancer, and FcγRI is expressed on human monocytes, macrophages, and IFN-γ activated granulocytes. We investigated phagocytosis and cytolysis of cultured human breast cancer cells by human monocyte-derived macrophages (MDM) mediated by BsAb MDX-210, its partially humanized derivative (MDX-H210), and its parent MoAb 520C9 (anti-HER-2/neu) under various conditions.

Materials and Methods: Purified monocytes were cultured with GM-CSF, M-CSF, or no cytokine for five or six days. Antibody dependent cellular phagocytosis (ADCP) and cytolysis (ADCC) assays were performed with the MDM and HER-2/neu positive target cells (SK-BR-3). ADCP was measured by two-color fluorescence flow cytometry using PKH2 (green fluorescent dye) and phycoerythrin-conjugated (red) monoclonal antibodies (MoAb) against human CD14 and CD11b. ADCC was measured with a non-radioactive LDH detection kit.

Results: Both BsAb MDX-210 (via FcγRI) and MoAb 520C9 (mouse IgG1, via FcγRII) mediated similar levels of ADCP and ADCC. ADCP mediated by BsAb MDX-H210 was identical to that mediated by BsAb MDX-210. Confocal microscopy demonstrated that dual-labeled cells represented true phagocytosis. Both ADCP and ADCC were higher when MDM were pre-incubated with GM-CSF than when incubated with M-CSF.

Conclusions: BsAb MDX-210 is as active in vitro as the parent MoAb 520C9 in inducing both phagocytosis and cytolysis of MDM. MDX-210 and its partially humanized derivative, MDX-H210, mediated similar levels of ADCP. GM-CSF appears to superior to M-CSF in inducing MDM-mediated ADCC and ADCP. These studies support the ongoing clinical investigations of BsAb MDX-210 and its partially humanized derivative.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. King CR, Kraus MH, Aaronson SA: Amplification of a novel v-erbB-related gene in a human mammary carcinoma. Science 229: 974–976, 1985

    Google Scholar 

  2. Ring DB, Kassel JA, Hsieh-Ma ST, Bjorn MJ, Tringale F, Eaton AM, et al.: Distribution and physical properties of BCA200, aMr 200,000 glycoprotein selectively associated with human breast cancer. Cancer Res 49: 3070–3080, 1989

    Google Scholar 

  3. Kraus MH, Popescu NC, Amsbaugh SC, King CR: Overexpression of the EGF receptor-related proto-oncogene erbB-2 in human mammary tumor cell lines by different molecular mechanisms. EMBO J 6: 605–610, 1987

    Google Scholar 

  4. Ullrich A, Schlessinger J: Signal transduction by receptors with tyrosine kinase activity. Cell 61: 203–212, 1990

    Google Scholar 

  5. Yarden Y, Ullrich A: Growth factor receptor tyrosine kinases. Ann Rev Biochem 57: 443–478, 1988

    Google Scholar 

  6. Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, et al.: Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 244: 707–712, 1989

    Google Scholar 

  7. Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL: Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235: 177–182, 1987

    Google Scholar 

  8. Ring DB, Clark R, Saxena A: Identity of BCA200 and cerbB-2 indicated by reactivity of monoclonal antibodies with recombinant c-erbB-2. Mol Immunol 28: 915–917, 1991

    Google Scholar 

  9. Guyre PM, Graziano RF, Vance BA, Morganelli PM, Fanger MW: Monoclonal antibodies that bind to distinct epitopes on Fc RI are able to trigger receptor function. J Immunol 143: 1650–1655, 1989

    Google Scholar 

  10. Wallace PK, Howell AL, Fanger MW: Role of Fc receptors in cancer and infectious disease. J Leukocyte Biol 55: 816–826, 1994

    Google Scholar 

  11. Fanger MW, Morganelli PM, Guyre PM: Bispecific antibodies. Critical Rev Immunol 12: 101–124, 1992

    Google Scholar 

  12. Fanger MW, Morganelli PM, Guyre PM: Use of bispecific antibodies in the therapy of tumors. Cancer Treat Res 68: 181–194, 1993

    Google Scholar 

  13. Valone FH, Kaufman PA, Guyre PM, Lewis LD, Memoli V, Ernstoff MS, et al.: Clinical trials of bispecific antibody MDX-210 inwomen with advanced breast or ovarian cancer that overexpresses HER-2/neu. J Hematother 4: 471–475, 1995

    Google Scholar 

  14. van de Winkel JG, Capel PJ: Human IgG Fc receptor heterogeneity: molecular aspects and clinical implications. Immunol Today 14: 215–221, 1993

    Google Scholar 

  15. Keler T, Graziano RF, Mandal A, Wallace PK, Fisher J, Guyre PM, et al.: Bispecific antibody-dependent cellular cytotoxicity of HER-2/neu-overexpressing tumor cells by Fc receptor type I-expressing effector cells. Cancer Res 57: 4008–4014, 1997

    Google Scholar 

  16. Valone FH, Kaufman PA, Guyre PM, Lewis LD, Memoli V, Deo Y, et al.: Phase Ia/Ib trial of bispecific antibody MDX-210 in patients with advanced breast or ovarian cancer that overexpresses the proto-oncogene HER-2/neu. J Clin Oncol 13: 2281–2292, 1995

    Google Scholar 

  17. Posey J, Verma U, Marshall JL, Raspet R, Mazumder A, Hawkins MJ: Pilot trial of MDX-H210 and GM-CSF for patients with advanced erbB-2 positive malignancies. Proceedings of the American Association for Cancer Research 37: p. 1136, 1996

    Google Scholar 

  18. Kaufmann PA, Lewis LD, Barth RJ, Guyre PM, Wallace PK, Memoli VA, et al.: Bispecific antibody 520C9x22 (MDX-210) and interferon gamma is an immunologically active treatment for patients with metastatic adenocarcinomas that overexpress HER-2/NEU. Breast Cancer Res Treat 41: p. 246a, 1996

    Google Scholar 

  19. Glennie MJ, McBride HM, Worth AT, Stevenson GT: Preparation and performance of bispecific F.ab0/2 antibody containing thioether-linked Fab0 fragments. J Immunol 139: 2367–2375, 1987

    Google Scholar 

  20. Anderson CL, Guyre PM, Whitin JC, Ryan DH, Looney RJ, Fanger MW: Monoclonal antibodies to Fc receptors for IgG on human mononuclear phagocytes. Antibody characterization and induction of superoxide production in a monocyte cell line. J Biol Chem 261: 12856–12864, 1986

    Google Scholar 

  21. Sone S, Nii M: Monocytes. In: Tokunaga T, Yoshida T, Akagawa K (eds) Makurofaji Jikken Manyuaru. Kodansya, Tokyo; 1994, pp. 4–7

    Google Scholar 

  22. Backman KA, Guyre PM: γ-interferon inhibits Fc receptor II-mediated phagocytosis of tumor cells by human macrophages. Cancer Res 54: 2456–2461, 1994

    Google Scholar 

  23. Ely P, Wallace PK, Givan AL, Graziano RF, Guyre PM, Fanger MW: Bispecific-armed, interferon-primed macrophage-mediated phagocytosis of malignant non-Hodgkin's lymphoma. Blood 87: 3813–3821, 1996

    Google Scholar 

  24. Munn DH, Cheung NK: Phagocytosis of tumor cells by human monocytes cultured in recombinant macrophage colony-stimulating factor. J Exp Med 172: 231–237, 1990

    Google Scholar 

  25. Munn DH, McBride M, Cheung NK: Role of low-affinity Fc receptors in antibody-dependent tumor cell phagocytosis by human monocyte-derived macrophages. Cancer Res 51: 1117–1123 1991

    Google Scholar 

  26. Stockmeyer B, Valerius T, Repp R, Heijnen IAFM, Buhring HS, Deo YM, et al.: Preclinical studies with FcR bispecific antibodies and granulocyte colony-stimulating factorprimed neutrophils as effector cells against HER-2/neu overexpressing breast cancer. Cancer Res 57: 696–701, 1997

    Google Scholar 

  27. Looney RJ, Abraham GN, Anderson CL:Humanmonocytes and U937 cells bear two distinct Fc receptors for IgG. J Immunol 136: 1641–1647, 1986

    Google Scholar 

  28. Liesveld J, Frediani K, Winslow J, Duerst R, Abboud C: Cytokine effects and role of adhesive proteins and Fc receptors in human macrophage-mediated antibody dependent cellular cytotoxicity. J Cell Biochem 45: 381–390, 1991

    Google Scholar 

  29. Graziano R, Tempest P, White P, Keler T, Deo Y, Ghebremearian H, et al.: Construction and characterization of a humanized anti--Ig receptor Type I (Fc RI) monoclonal antibody. J Immunol 155: 4996–5002, 1995

    Google Scholar 

  30. Repp R, Valerius T, Wieland G, Becker W, Steininger H, Deo Y, et al.: G-CSF-stimulated PMN in immunotherapy of breast cancer with a bispecific antibody to Fc RI and to HER-2/neu (MDX-210). J Hematother 4: 415–421, 1995

    Google Scholar 

  31. Munn DH, Cheung NK: Antibody-independent phagocytosis of tumor cells by human monocyte-derived macrophages cultured in recombinant macrophage colony-stimulating factor. Cancer Immunol and Immunother 41: 46–52, 1995

    Google Scholar 

  32. Deo Y, Sundarapandiyan K, Keler T, Wallace P, Graziano R: Bispecific molecules directed to the Fc Receptor for IgA (Fc_RI, DC89) and tumor antigens efficiently promote cellmediated cytotoxicity of tumor targets in whole blood. J Immunol 160: 1677–1686, 1998

    Google Scholar 

  33. Vadhan-Raj S, Keating M, LeMaistre A, Hittelman W, Mc-Credie K, Trujillo J, et al.: Effects of recombinant human granulocyte-macrophage colony stimulating factor in patients with myelodysplastic syndromes. N Engl J Med 317: 1545–1552, 1987

    Google Scholar 

  34. Fanger MW, Shen L, Graziano RF, Guyre PM: Cytotoxicity mediated by human Fc receptors for IgG. Immunol Today 10: 92–99, 1989

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Breast Cancer Program, Lombardi Cancer Center, Georgetown University Medical Center, Washington DC

    Michiaki Watanabe, Charuwan Akewanlop & Daniel F. Hayes

  2. Department of Microbiology, Dartmouth Medical School, Lebanon, NH

    Paul K. Wallace

  3. Medarex, Annandale, NJ

    Tibor Keler & Yashwant M. Deo

Authors
  1. Michiaki Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Paul K. Wallace
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tibor Keler
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yashwant M. Deo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Charuwan Akewanlop
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Daniel F. Hayes
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Watanabe, M., Wallace, P.K., Keler, T. et al. Antibody dependent cellular phagocytosis (ADCP) and antibody dependent cellular cytotoxicity (ADCC) of breast cancer cells mediated by bispecific antibody, MDX-210. Breast Cancer Res Treat 53, 199–207 (1999). https://doi.org/10.1023/A:1006145507567

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006145507567

Search

Navigation