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Journal of Histochemistry and Cytochemistry, Vol. 49, 911-918, July 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Lysis of Prostate Carcinoma Cells by Trifunctional Bispecific Antibodies ({alpha}EpCAM x {alpha}CD3)

Rainer Riesenberga, Alexander Buchnera, Heike Pohlaa, and Horst Lindhoferb
a Laboratory for Tumorimmunology, Department of Urology, Ludwig-Maximilians-University, Klinikum Grosshadern, Munich, Germany
b Clinical Cooperation group "Bispecific Antibodies" at the Department of Otorhinolaryngology, Ludwig-Maximilians-University, and the Institute of Clinical Molecular Biology, GSF–National Research Center for Environment and Health, Munich, Germany

Correspondence to: Rainer Riesenberg, Labor für Tumorimmunologie, Urologische Klinik, Klinikum Grosshadern, Marchioninistr. 23, D-81377 Munich, Germany. E-mail: riesenberg@life.med.uni-muenchen.de

Bispecific monoclonal antibodies (bsAbs) are a promising immunotherapeutic option for treatment of cancer, especially in situations of minimal residual disease. The combination of an anti-CD3 and anti-tumor-associated antigen antibody redirects cytotoxic T-lymphocytes towards malignant cells. Using a trifunctional bispecific antibody against EpCAM x CD3, that additionally activates Fc{gamma}R+ accessory cells via its Fc region, we investigated the interaction between three EpCAM+ prostate carcinoma cell lines and peripheral blood mononuclear cells (PBMCs) of healthy donors and patients with prostate carcinoma (PC). Visualization was performed by double immunocytochemical methods and computerized sequential video microscopy. Tumor cells and PBMCs supplemented with {alpha}EpCAM x {alpha}CD3 in 16-well chamber slides resulted in lysis of tumor cells within 1–3 days without any differences between patient and healthy donor PBMCs. The characteristic necrotic way of tumor cell killing (rounding, swelling, disrupting) could be observed in computerized sequences of video frames. Simultaneously, we could not reveal any form of apoptotic signal using three different apoptotic markers (TUNEL, M30 cyto death, anti-active caspase 3). Within the first 48 hr we observed typical PBMC cluster formation with increasing cell proliferation. PBMCs surrounding the tumor cells were not dominated by CD4+, CD8+, or CD14+ cells. Lymphocytes with pore-forming perforin proteins concentrated towards the tumor target cells. Our combination of double immunocytochemical and computerized video microscopic techniques may serve as an important improvement of validity of cell–cell interaction experiments using in vitro models. (J Histochem Cytochem 49:911–917, 2001)

Key Words: prostate carcinoma, bispecific antibody, double immunocytochemistry, computerized sequential, microscopy


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