Bacillus Calmette-Guérin interacts with the carboxyl-terminal heparin binding domain of fibronectin: implications for BCG-mediated antitumor activity.
Intravesical bacillus Calmette-Guérin (BCG) has been shown to be an effective treatment for superficial transitional cell carcinoma of the bladder. The mechanisms by which BCG achieves this effect remain unclear. Reports have attributed an important role to fibronectin both in the initial attachment of BCG to bladder surfaces and in the limitation of tumor cell motility. In the present study, using limited protease cathepsin B degradation followed by Western blot analyses with antibodies to various domains of the fibronectin molecule, we showed that BCG appears to bind to fibronectin near the carboxyl terminal and adjacent to the heparin binding domain. Furthermore a 51-chromium release assay with human bladder cancer cell line T24 as target cells and lymphokine activated killer (LAK) cells as effector cells showed that fibronectin was needed for tumor cytotoxicity by the LAK cells. By using antibodies and peptides to various domains of the fibronectin molecule, the heparin binding domain, but not the cell binding domain, carboxyl terminal region, or the amino terminal region of the fibronectin molecule, was identified as essential to tumor cell lysis by the LAK cells. Flow cytometric analysis showed that both peripheral blood lymphocytes and the LAK cells express fibronectin receptors VLA-3, VLA-4 and VLA-5 on their surfaces. However, the numbers of receptors are not significantly different in the two cell populations. We conclude that, by binding near the carboxyl terminal region and adjacent to the heparin-binding domain of the fibronectin molecule, BCG may protect this region of the molecule from tumor proteases, and may thus allow the antitumor activity of the host immune cells to take place.
Cheng
D L
DL
Department of Urology, Mount Sinai School of Medicine, New York, New York 10029.
Shu
W P
WP
Choi
J C
JC
Margolis
E J
EJ
Droller
M J
MJ
Liu
B C
BC
D001500:BCG Vaccine / Q000378:metabolism* / Q000627:therapeutic use; D001665:Binding Sites; D002295:Carcinoma, Transitional Cell / Q000378:metabolism* / Q000628:therapy; D003602:Cytotoxicity, Immunologic; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D015979:Killer Cells, Lymphokine-Activated / Q000502:physiology; D007962:Leukocytes / Q000378:metabolism; D011485:Protein Binding; D017469:Receptors, Fibronectin / Q000096:biosynthesis; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000628:therapy
D001500:BCG Vaccine; D005353:Fibronectins; D017469:Receptors, Fibronectin; D006493:Heparin