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Stable expression of chimeric anti-CD3 receptors on mammalian cells for stimulation of antitumor immunity

Cancer Gene Therapy volume 10, pages 779–790 (2003)Cite this article

Abstract

Expression of CD80 or CD86 costimulatory molecules on tumor cells can produce rejection of immunogenic but not poorly immunogenic tumors. We have previously shown that anti-CD3 single-chain antibodies expressed on the surface of cells can directly activate T cells. We therefore investigated whether anti-CD3 “receptors” could enhance CD86-mediated rejection of poorly immunogenic tumors. Expression of anti-CD3 receptors on cells was increased by introduction of membrane-proximal “spacer” domains containing glycosylation sites between the single-chain antibody and the transmembrane domain of the chimeric receptors. Removal of glycosylation sites in the spacer reduced surface expression due to increased shedding of chimeric receptors from the cell surface. Induction of T-cell proliferation by anti-CD3 receptors did not correlate with the expression level of chimeric protein, but rather depended on the physical properties of the spacer. Anti-CD3 receptors effectively induced T-cell cytotoxicity, whereas coexpression with CD80 or CD86 was required for generating T-cell proliferation and IL-2 secretion. Although expression of CD86 did not significantly delay the growth of poorly immunogenic B16-F1 tumors, expression of anti-CD3 receptors with CD86 produced complete tumor rejections in 50% of mice and induced significant protection against wild-type B16-F1 tumor cells. Our results show that spacer domains can dramatically influence the surface expression and the biological activity of chimeric antibody receptors. The strong antitumor activity produced by anti-CD3 receptors and CD86 on tumor cells indicates that this strategy may be beneficial for the gene-mediated therapy of poorly immunogenic tumors.

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Abbreviations

2C11:

anti-CD3 antibody

γ 1 :

H–CH2–CH3 region of human IgG1

AFP:

alpha fetoprotein

phOx:

4-ethoxymethylene-2-phenyl-2-oxazolin-5-one

scFv:

single-chain antibody

TM:

transmembrane domain

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Acknowledgements

We appreciate the technical assistance of Wesley Roy Balasubramanian. This study was supported by grants from the National Science Council, Taipei, Taiwan (NSC90-2318-B001-006-M51 and NSC91-3112-P001-026-Y).

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Author notes

  1. Kuang-Wen Liao and Bing-Mae Chen: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Biomedical Sciences, Academia Sinica, Taipei, 11529, Taiwan, Republic of China

    Kuang-Wen Liao, Bing-Mae Chen, Tang-Bi Liu, Shey-Cherng Tzou, Kai-Feng Lin, Chien-I Su & Steve R Roffler

  2. Institute of Molecular Biology, Academia Sinica, Taipei, 11529, Taiwan, Republic of China

    Ya-Min Lin

  3. Department of Life Sciences, National Tsing Hua University, Hsin-Chu, 30043, Taiwan, Republic of China

    Chien-I Su

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  1. Kuang-Wen Liao
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Correspondence to Steve R Roffler.

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Liao, KW., Chen, BM., Liu, TB. et al. Stable expression of chimeric anti-CD3 receptors on mammalian cells for stimulation of antitumor immunity. Cancer Gene Ther 10, 779–790 (2003). https://doi.org/10.1038/sj.cgt.7700637

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