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The in vitro antitumor effect and in vivo tumor-specificity distribution of human-mouse chimeric antibody against transferrin receptor

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Abstract

Transferrin receptor (TfR/CD71) deserves attention as a selective target for cancer therapy due to its higher expression in tumors versus normal tissues. Also, it has been shown the mouse-derived monoclonal antibody against TfR can significantly inhibit the proliferation of tumor cells. Through constructing the chimeric antibody against TfR, the antigenicity of antibody can be weakened, and most importantly, the antitumor effect of antibody can be strengthened by the introduction of the human Fc fragment. In previous studies, we successfully constructed the human-mouse chimeric antibody against TfR (D2C) and demonstrated that its Fab fragment could specially recognize the TfR on the surface of target cells. In this study, through labeling the chimeric antibody D2C with 125I, we calculated the affinity constant (Ka) of 9.34–9.62×109 l/mol for this antibody according to the Scatchard drawing method. Moreover, in vivo studies in nude mice-bearing human liver cancer (SMMC-7721) xenografts have shown that the radioactivity distribution ratio of 131I-D2C on T/NT was 2–14:1 or 3–21:1 on the seventh day after intraperitoneal or intratumoral injection of 131I-labeled D2C (131I-D2C). These evidences indicated that the in vivo distribution of D2C display the characteristics of certain tumor-specificity localization. In vitro studies, D2C can induce the apoptosis of K562 through the mitochondria death pathway and arrest the cell at G1 phase, as determined by cell cycle analysis. Using the human tumor cells (K562, CEM, and SMMC-7721) expressing TfR as target cells, and normal human PBMC as effector cells, Fc fragment of D2C can perform both the antibody-dependent cell-mediated cytotoxicity and the complement-dependent cytotoxicity. Together, it was demonstrated that the D2C display a tumor-specificity distribution, and has a strong antitumor effect. Thus, it has the potential therapeutic significance.

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Abbreviations

Ka:

Affinity constant

CDC:

Complement-dependent cytotoxicity

ADCC:

Antibody-dependent cell-mediated cytotoxicity

NHL:

Non-Hodgkin’s lymphoma

Tf:

Transferrin

TfR:

Transferrin receptor

FACS:

Fluorescence-activated cell sorting

SPECT:

Single photon emission computed tomography

D2C:

Human-mouse chimeric antibody against TfR

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PBMC:

Peripheral blood mononuclear cell

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Acknowledgements

This project was supported by National Key and Basic Research Development Program (no. 2002CB513109). We thank Prof. Gong Feili for helpful discussions and critical reading of the manuscript and many cited colleagues for the reagents supplied, and Zhang Yue, Yang Jing and Shao Jingfang for assistance with cell culture and maintenance.

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Authors and Affiliations

  1. Laboratory of Molecular and Immuno-Pharmacology, Department of Pharmacology and Immunology, Tongji Medical College, Huazhong University of Science and Technology, No. 13, Hongkong Road, 430030, Wuhan, People’s Republic of China

    Ye Qing, Zhu Huifen, Lei Ping, Zhao Xiaorong, Chao Liming, Xiao Daiwen, Huang Yu, Xing Wei, Fang Min, Feng Zuohua & Shen Guanxin

  2. Department of Pathobiology and Physiology, Medicine and Life Science College of Jianghan University, 430056, Wuhan, People’s Republic of China

    Ye Qing, Wang Shuo & Liu Lijiang

  3. Department of Urology, Tongji Hospital of Huazhong University of Science and Technology, 430030, Wuhan, People’s Republic of China

    Wang Zhihua

Authors
  1. Ye Qing
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  2. Wang Shuo
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  3. Wang Zhihua
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  4. Zhu Huifen
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  7. Zhao Xiaorong
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  9. Xiao Daiwen
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  10. Huang Yu
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  11. Xing Wei
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  12. Fang Min
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  13. Feng Zuohua
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  14. Shen Guanxin
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Corresponding author

Correspondence to Shen Guanxin.

Additional information

Ye Qing and Wang Shuo contributed equally to this work.

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Qing, Y., Shuo, W., Zhihua, W. et al. The in vitro antitumor effect and in vivo tumor-specificity distribution of human-mouse chimeric antibody against transferrin receptor. Cancer Immunol Immunother 55, 1111–1121 (2006). https://doi.org/10.1007/s00262-005-0105-7

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  • DOI: https://doi.org/10.1007/s00262-005-0105-7

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