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Cancer Immunology, Immunotherapy

, Volume 58, Issue 11, pp 1887–1896 | Cite as

Breast tumor cells isolated from in vitro resistance to trastuzumab remain sensitive to trastuzumab anti-tumor effects in vivo and to ADCC killing

  • Timothy E. Kute
  • Lori Savage
  • John R. StehleJr
  • Jung W. Kim-Shapiro
  • Michael J. Blanks
  • James Wood
  • James P. Vaughn
Original article

Abstract

An understanding of model systems of trastuzumab (Herceptin) resistance is of great importance since the humanized monoclonal antibody is now used as first line therapy with paclitaxel in patients with metastatic Her2 overexpressing breast cancer, and the majority of their tumors has innate resistance or develops acquired resistance to the treatment. Previously, we selected trastuzumab-resistant clonal cell lines in vitro from trastuzumab-sensitive parental BT-474 cells and showed that cloned trastuzumab-resistant cell lines maintain similar levels of the extracellular Her2 receptor, bind trastuzumab as efficiently as the parental cells, but continue to grow in the presence of trastuzumab and display cell cycle profiles and growth rates comparable to parental cells grown in the absence of trastuzumab (Kute et al. in Cytometry A 57:86–93, 2004). We now show that trastuzumab-resistant and trastuzumab-sensitive cells both surprisingly display trastuzumab-mediated growth inhibition in athymic nude mice. This demonstrates that resistance developed in vitro is not predictive of resistance in vivo. The observation that in vitro resistant cells are sensitive to trastuzumab in vivo could be explained by antibody dependant cellular cytotoxicity (ADCC). Therefore, both parental and trastuzumab-resistant cells were assayed for ADCC in real time on electroplates with and without trastuzumab in the presence of a natural killer cell line (NK-92), and granulocyte or mononuclear cellular fractions isolated from human peripheral blood. Mononuclear cells and NK-92 cells were more effective in killing both parental and trastuzumab-resistant cells in the presence of trastuzumab. Both trastuzumab-resistant cells and trastuzumab-sensitive cells showed similar susceptibility to ADCC despite displaying divergent growth responses to trastuzumab. The granulocyte fraction was able to kill these cells with equal efficacy in the presence or absence of trastuzumab. These results support a model of trastuzumab tumor cell killing in vivo mediated primarily by ADCC from the mononuclear fraction of innate immune cells and suggest that in the clinical setting not only should changes in signaling transduction pathways be studied in acquired tumor resistance to trastuzumab, but also mechanisms by which tumors impede immune function should be evaluated.

Keywords

ADCC Trastuzumab Herceptin Breast cancer Resistance BT-474 

Notes

Acknowledgments

This work was performed from a grant from the Vaughn Jordon Foundation and from support of the pathology department and Wake Forest University Cancer Center. We would also like to thank Ms Debra Holder who help with the editing of the manuscript and Dr. Mark Willingham and Dr. Zheng Cui who provided comments and suggestions. Finally, I would like to thank Dr. Greg Russell for his help with the statistics.

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Timothy E. Kute
    • 1
  • Lori Savage
    • 1
  • John R. StehleJr
    • 1
  • Jung W. Kim-Shapiro
    • 1
  • Michael J. Blanks
    • 1
  • James Wood
    • 2
  • James P. Vaughn
    • 2
  1. 1.Department of PathologyWake Forest UniversityWinston-SalemUSA
  2. 2.Department of Cancer BiologyWake Forest University School of MedicineWinston-SalemUSA

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