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Specific binding of bacillus Calmette-Guérin to urothelial tumor cells in vitro

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Summary

Intravesical immunotherapy with bacillus Calmette-Guérin (BCG) against recurrences of superficial bladder cancer and carcinoma in situ is a highly effective regimen in urology. Despite intensive efforts to clarify the immunological mechanisms of the most successful immunotherapy known today, the cellular mechanism of its antitumor activity remains unknown. In our approach to elucidate the way of action of intravesical BCG, we applied an in vitro adhesion assay to investigate the interaction of radiolabeled BCG with urothelial bladder-tumor cells. We demonstrated a BCG dose-dependent binding to bladder-tumor cell lines derived from tumors of different gradings. The binding of BCG is apparently specific, since competition experiments showed an inhibition by nonradioactive BCG but not by Escherichia coli. We also found that there was no difference between the binding of living or heat-killed mycobacteria. Control experiments showed only a low affinity of BCG for fibroblasts, smooth-muscle cells, and endothelial cells in comparison with the tumor cells. Furthermore, we investigated the role of fibronectin as an adhesion molecule that is also present in the bladder wall. We demonstrated that BCG was capable of binding to fibronectin-coated surfaces in a dose-dependent manner. However, competitive binding assays failed to reveal an inhibition of the binding of BCG to bladder-tumor cells by anti-fibronectin. Furthermore, binding was not influenced by soluble fibronectin. These data suggest that the in vitro attachment of BCG to bladder-tumor cells appears not to be mediated by fibronectin. In electron microscope studies an adhesion of BCG to bladder-tumor cells was observed after an incubation period of only 30 min. After 24 h of incubation, we saw in addition that tumor cell lines of all different gradings had phagocytosed the mycobacteria. The phagocytosed mycobacteria within vacuoles were in various states of structural integrity ranging from completely intact to almost fully disintegrated. In contrast, fibroblasts were incapable of engulfing BCG. We conclude that BCG can bind to bladder-tumor cells in a specific manner and can be phagocytosed by these cells. Tumor cells of all gradings showed this behavior, but fibroblasts did not. The specific interaction observed between BCG and bladder-tumor cells of all gradings might be an important step in the development of antitumor defense mechanisms in bladder cancer patients.

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Authors and Affiliations

  1. Department of Urology, Medical University of Lübeck, D-23538, Lübeck, Germany

    B. Schneider, D. Jocham & A. Böhle

  2. Department of Immunology and Cell Biology, Forschungsinstitut Borstel, D-23845, Borstel, Germany

    A. Thanhäuser, H. Loppnow, H. -D. Flad & A. J. Ulmer

  3. Division of Clinical and Experimental Pathology, Forschungsinstitut Borstel, D-23845, Borstel, Germany

    E. Vollmer & J. Galle

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  1. B. Schneider
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  2. A. Thanhäuser
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  3. D. Jocham
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  4. H. Loppnow
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  8. A. J. Ulmer
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  9. A. Böhle
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Schneider, B., Thanhäuser, A., Jocham, D. et al. Specific binding of bacillus Calmette-Guérin to urothelial tumor cells in vitro. World J Urol 12, 337–344 (1994). https://doi.org/10.1007/BF00184116

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  • DOI: https://doi.org/10.1007/BF00184116

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