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Co-expression of α(1,3)galactosyltransferase and Bacillus thuringiensis PIPLC enhances hyperacute rejection of tumor cells

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Abstract

The use of α(1,3)galactosyltransferase (αGT) as a method of inducing hyperacute rejection of tumors has been gaining interest recently. However, the approach is based in part on the sensitivity of each tumor line to the effects of complement lysis. Tumors expressing complement resistance factors such as membrane cofactor (CD46), decay accelerating factor (CD55) and protectin (CD59) have been shown to be more resistant to complement mediated lysis. Anchored to the membrane by a glycosylphosphoinositol moiety (GPI-anchored), CD55 and CD59 can be cleaved by Bacillus thuringiensis phosphatidylinositol-specific phospholipase C (PIPLC). Complement resistant A549 human lung carcinoma cells were engineered to express both the murine αGT gene and the B. thuringiensis PIPLC gene to alleviate complement resistance and enhance αgal-mediated cancer killing. The PIPLC native signal sequence was replaced with the human epidermal growth factor signal sequence, EGFssPIPLC, to induce secretion from A549. Expression of EGFssPIPLC resulted in complete removal of CD55 and CD59 while sparing the non-GPI-anchored CD46. Results demonstrated that A549 cells transduced with two recombinant retroviral vectors carrying the αGT and EGFssPIPLC genes expressed high levels of αgal epitope and exhibited a 5-fold increase in sensitivity to anti-αgal mediated complement lysis.

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Acknowledgements

The authors thank Dr. Ming Tsai of Ohio State University for providing the B. thuringiensis PIPLC gene and Dr. Mary Roberts of Boston college for her helpful discussions. This work was funded by a grant from the Department of Defense Army Breast Cancer Foundation (#DAMD17-00-1-0292) and the Susan G. Komen Breast Cancer Foundation (#99–3215).

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Authors and Affiliations

  1. Iowa Cancer Research Foundation, 11043 Aurora Avenue, Urbandale, IA, 50322, USA

    Daniel J. Hellrung, Serguei Kisselev & Charles J. Link

  2. Iowa State University, Ames, IA, USA

    Daniel J. Hellrung & Charles J. Link

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  1. Daniel J. Hellrung
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Correspondence to Charles J. Link.

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Hellrung, D.J., Kisselev, S. & Link, C.J. Co-expression of α(1,3)galactosyltransferase and Bacillus thuringiensis PIPLC enhances hyperacute rejection of tumor cells. Cancer Immunol Immunother 56, 25–34 (2007). https://doi.org/10.1007/s00262-006-0163-5

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