Bispecific antibody-targeted phagocytosis of HER-2/neu expressing tumor cells by myeloid cells activated in vivo

Paul K. Wallace; Peter A. Kaufman; Lionel D. Lewis; Tibor Keler; Alice L. Givan; Jan L. Fisher; Mary G. Waugh; Andrea E. Wahner; Paul M. Guyre; Michael W. Fanger; Marc S. Ernstoff

doi:10.1016/S0022-1759(00)00350-1

Bispecific antibody-targeted phagocytosis of HER-2/neu expressing tumor cells by myeloid cells activated in vivo

Paul K. Wallace, Peter A. Kaufman, Lionel D. Lewis, Tibor Keler, Alice L. Givan, Jan L. Fisher, Mary G. Waugh, Andrea E. Wahner, Paul M. Guyre, Michael W. Fanger, Marc S. Ernstoff

Medical Oncology

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

Studies from our laboratory and others have established that both mononuclear phagocytes and neutrophils mediate very efficient cytotoxicity when targeted through Fc receptors using a suitable monoclonal or bispecific antibody (BsAb). Cross-linking an Fc receptor for IgG (FcγR) triggers multiple anti-tumor activities including superoxide generation, cytokine and enzyme release, phagocytosis and antibody-dependent cellular cytotoxicity (ADCC). In this report, using unfractionated leukocytes and two color flow cytometric analysis, we describe the phagocytic capacity of peripheral blood polymorphonuclear cells (PMN) and monocytes isolated from patients enrolled in a phase I clinical trial of MDX-H210 given in combination with IFNγ. MDX-H210 is a BsAb targeting the myeloid trigger molecule FcγRI and the HER-2/neu proto-oncogene product overexpressed on a variety of adenocarcinomas. In this trial, cohorts of patients received escalating doses of MDX-H210 3 times per week for 3 weeks. Interferon-γ (IFNγ) was given 24 h prior to each BsAb infusion. Our results demonstrate that monocytes from these patients were inherently capable of phagocytosing the HER-2/neu positive SK-BR-3 cell line and that addition of MDX-H210 into the assay significantly enhanced the number of targets phagocytosed. Two days after administration of an immunologically active dose of MDX-H210 (10 mg/m2), monocytes from these patients were able to phagocytose greater amounts of target cell material, indicating that these cells remained armed with functionally sufficient BsAb for at least 48 h. PMN from these patients very efficiently mediated phagocytosis through FcγRI after being treated with IFNγ, but not before. We conclude that phagocytosis is not only an efficient mechanism of myeloid cell-mediated cytotoxicity, but may also be a mechanism by which antigens from phagocytosed cells can enter a professional antigen presenting cell for processing and presentation.

Original language	English (US)
Pages (from-to)	167-182
Number of pages	16
Journal	Journal of Immunological Methods
Volume	248
Issue number	1-2
DOIs	https://doi.org/10.1016/S0022-1759(00)00350-1
State	Published - Feb 1 2001

Keywords

ADCC
Bispecific antibody
Fc receptor for IgG
HER-2/neu
Phagocytosis

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1016/S0022-1759(00)00350-1

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Wallace, P. K., Kaufman, P. A., Lewis, L. D., Keler, T., Givan, A. L., Fisher, J. L., Waugh, M. G., Wahner, A. E., Guyre, P. M., Fanger, M. W., & Ernstoff, M. S. (2001). Bispecific antibody-targeted phagocytosis of HER-2/neu expressing tumor cells by myeloid cells activated in vivo. Journal of Immunological Methods, 248(1-2), 167-182. https://doi.org/10.1016/S0022-1759(00)00350-1

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title = "Bispecific antibody-targeted phagocytosis of HER-2/neu expressing tumor cells by myeloid cells activated in vivo",

abstract = "Studies from our laboratory and others have established that both mononuclear phagocytes and neutrophils mediate very efficient cytotoxicity when targeted through Fc receptors using a suitable monoclonal or bispecific antibody (BsAb). Cross-linking an Fc receptor for IgG (FcγR) triggers multiple anti-tumor activities including superoxide generation, cytokine and enzyme release, phagocytosis and antibody-dependent cellular cytotoxicity (ADCC). In this report, using unfractionated leukocytes and two color flow cytometric analysis, we describe the phagocytic capacity of peripheral blood polymorphonuclear cells (PMN) and monocytes isolated from patients enrolled in a phase I clinical trial of MDX-H210 given in combination with IFNγ. MDX-H210 is a BsAb targeting the myeloid trigger molecule FcγRI and the HER-2/neu proto-oncogene product overexpressed on a variety of adenocarcinomas. In this trial, cohorts of patients received escalating doses of MDX-H210 3 times per week for 3 weeks. Interferon-γ (IFNγ) was given 24 h prior to each BsAb infusion. Our results demonstrate that monocytes from these patients were inherently capable of phagocytosing the HER-2/neu positive SK-BR-3 cell line and that addition of MDX-H210 into the assay significantly enhanced the number of targets phagocytosed. Two days after administration of an immunologically active dose of MDX-H210 (10 mg/m2), monocytes from these patients were able to phagocytose greater amounts of target cell material, indicating that these cells remained armed with functionally sufficient BsAb for at least 48 h. PMN from these patients very efficiently mediated phagocytosis through FcγRI after being treated with IFNγ, but not before. We conclude that phagocytosis is not only an efficient mechanism of myeloid cell-mediated cytotoxicity, but may also be a mechanism by which antigens from phagocytosed cells can enter a professional antigen presenting cell for processing and presentation.",

keywords = "ADCC, Bispecific antibody, Fc receptor for IgG, HER-2/neu, Phagocytosis",

author = "Wallace, {Paul K.} and Kaufman, {Peter A.} and Lewis, {Lionel D.} and Tibor Keler and Givan, {Alice L.} and Fisher, {Jan L.} and Waugh, {Mary G.} and Wahner, {Andrea E.} and Guyre, {Paul M.} and Fanger, {Michael W.} and Ernstoff, {Marc S.}",

note = "Funding Information: Supported in part by grants from Amgen (PKW), The American Cancer Societies Institute Research Grant (IRG-82-003-16, PKW), The National Institute for Heath (AI-34478, PKW, PMG, MWF), (CA-65953 MSE, PAK, JLF), The Pharmaceutical Manufacturers Research Foundation (LDL) and a Clinical Research Grant from Medarex, Inc., Annandale, NJ. Flow cytometry and laser scanning confocal microscopy were performed at Dartmouth Medical School in the Herbert C. Englert Cell Analysis Laboratory which was established with equipment grants from the Fannie E. Rippel Foundation and the NIH Shared Instrument Program and is supported in part by the core grant of the Norris Cotton Cancer Center (CA-23108). ",

year = "2001",

month = feb,

day = "1",

doi = "10.1016/S0022-1759(00)00350-1",

language = "English (US)",

volume = "248",

pages = "167--182",

journal = "Journal of Immunological Methods",

issn = "0022-1759",

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T1 - Bispecific antibody-targeted phagocytosis of HER-2/neu expressing tumor cells by myeloid cells activated in vivo

AU - Wallace, Paul K.

AU - Kaufman, Peter A.

AU - Lewis, Lionel D.

AU - Keler, Tibor

AU - Givan, Alice L.

AU - Fisher, Jan L.

AU - Waugh, Mary G.

AU - Wahner, Andrea E.

AU - Guyre, Paul M.

AU - Fanger, Michael W.

AU - Ernstoff, Marc S.

N1 - Funding Information: Supported in part by grants from Amgen (PKW), The American Cancer Societies Institute Research Grant (IRG-82-003-16, PKW), The National Institute for Heath (AI-34478, PKW, PMG, MWF), (CA-65953 MSE, PAK, JLF), The Pharmaceutical Manufacturers Research Foundation (LDL) and a Clinical Research Grant from Medarex, Inc., Annandale, NJ. Flow cytometry and laser scanning confocal microscopy were performed at Dartmouth Medical School in the Herbert C. Englert Cell Analysis Laboratory which was established with equipment grants from the Fannie E. Rippel Foundation and the NIH Shared Instrument Program and is supported in part by the core grant of the Norris Cotton Cancer Center (CA-23108).

PY - 2001/2/1

Y1 - 2001/2/1

N2 - Studies from our laboratory and others have established that both mononuclear phagocytes and neutrophils mediate very efficient cytotoxicity when targeted through Fc receptors using a suitable monoclonal or bispecific antibody (BsAb). Cross-linking an Fc receptor for IgG (FcγR) triggers multiple anti-tumor activities including superoxide generation, cytokine and enzyme release, phagocytosis and antibody-dependent cellular cytotoxicity (ADCC). In this report, using unfractionated leukocytes and two color flow cytometric analysis, we describe the phagocytic capacity of peripheral blood polymorphonuclear cells (PMN) and monocytes isolated from patients enrolled in a phase I clinical trial of MDX-H210 given in combination with IFNγ. MDX-H210 is a BsAb targeting the myeloid trigger molecule FcγRI and the HER-2/neu proto-oncogene product overexpressed on a variety of adenocarcinomas. In this trial, cohorts of patients received escalating doses of MDX-H210 3 times per week for 3 weeks. Interferon-γ (IFNγ) was given 24 h prior to each BsAb infusion. Our results demonstrate that monocytes from these patients were inherently capable of phagocytosing the HER-2/neu positive SK-BR-3 cell line and that addition of MDX-H210 into the assay significantly enhanced the number of targets phagocytosed. Two days after administration of an immunologically active dose of MDX-H210 (10 mg/m2), monocytes from these patients were able to phagocytose greater amounts of target cell material, indicating that these cells remained armed with functionally sufficient BsAb for at least 48 h. PMN from these patients very efficiently mediated phagocytosis through FcγRI after being treated with IFNγ, but not before. We conclude that phagocytosis is not only an efficient mechanism of myeloid cell-mediated cytotoxicity, but may also be a mechanism by which antigens from phagocytosed cells can enter a professional antigen presenting cell for processing and presentation.

AB - Studies from our laboratory and others have established that both mononuclear phagocytes and neutrophils mediate very efficient cytotoxicity when targeted through Fc receptors using a suitable monoclonal or bispecific antibody (BsAb). Cross-linking an Fc receptor for IgG (FcγR) triggers multiple anti-tumor activities including superoxide generation, cytokine and enzyme release, phagocytosis and antibody-dependent cellular cytotoxicity (ADCC). In this report, using unfractionated leukocytes and two color flow cytometric analysis, we describe the phagocytic capacity of peripheral blood polymorphonuclear cells (PMN) and monocytes isolated from patients enrolled in a phase I clinical trial of MDX-H210 given in combination with IFNγ. MDX-H210 is a BsAb targeting the myeloid trigger molecule FcγRI and the HER-2/neu proto-oncogene product overexpressed on a variety of adenocarcinomas. In this trial, cohorts of patients received escalating doses of MDX-H210 3 times per week for 3 weeks. Interferon-γ (IFNγ) was given 24 h prior to each BsAb infusion. Our results demonstrate that monocytes from these patients were inherently capable of phagocytosing the HER-2/neu positive SK-BR-3 cell line and that addition of MDX-H210 into the assay significantly enhanced the number of targets phagocytosed. Two days after administration of an immunologically active dose of MDX-H210 (10 mg/m2), monocytes from these patients were able to phagocytose greater amounts of target cell material, indicating that these cells remained armed with functionally sufficient BsAb for at least 48 h. PMN from these patients very efficiently mediated phagocytosis through FcγRI after being treated with IFNγ, but not before. We conclude that phagocytosis is not only an efficient mechanism of myeloid cell-mediated cytotoxicity, but may also be a mechanism by which antigens from phagocytosed cells can enter a professional antigen presenting cell for processing and presentation.

KW - ADCC

KW - Bispecific antibody

KW - Fc receptor for IgG

KW - HER-2/neu

KW - Phagocytosis

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Bispecific antibody-targeted phagocytosis of HER-2/neu expressing tumor cells by myeloid cells activated in vivo

Abstract

Keywords

ASJC Scopus subject areas

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