BCG-induced urinary cytokines inhibit microvascular endothelial cell proliferation.
OBJECTIVE
Angiogenesis is thought to depend on a net balance of molecules that inhibit or stimulate microvascular endothelial cells. A variety of molecules that affect angiogenesis are induced locally by the administration of intravesical bacille Calmette-Guerin (BCG) for superficial bladder cancer. We sought to determine whether BCG-induced urinary cytokines alter the effects of patient urine on assays of angiogenic activity.


METHODS
Patients undergoing BCG treatment provided urine samples before and at peak cytokine production times after BCG instillation. Fifty-four urine samples from 8 patients were analyzed by ELISA for a panel of molecules known to affect angiogenesis, and tested for angiogenic activity in human dermal microvascular endothelial cell (HDMEC) proliferation and migration assays. To assess the role of specific BCG-induced cytokines, urinary HDMEC proliferation assays were repeated in the presence of neutralizing antibodies to tumor necrosis factor-alpha (TNF-alpha), interferon-inducible protein-10 (IP-10), and/or interferon-gamma (IFN-gamma).


RESULTS
Urinary IFN-gamma, IP-10, TNF-alpha, and vascular endothelial growth factor (VEGF) were induced to nanogram/ml amounts by BCG treatment. While pre-BCG treatment urine samples minimally stimulated microvascular endothelial cell proliferation (+ 9%), post-BCG treatment urine became progressively inhibitory to endothelial cells (to -85%, p = 0.005) during weekly treatment courses. Neutralizing antibodies to TNF-alpha or to IP-10, either alone or in combination, greatly reduced this inhibitory effect.


CONCLUSIONS
Intravesical BCG induces a cytokine-rich urinary microenvironment that is inhibitory to human endothelial cells. Urinary cytokine profiles and assays of angiogenic inhibition may provide prognostically important information regarding BCG treatment outcomes.
Pavlovich
C P
CP
James Buchanan Brady Foundation, Department of Urology, The New York Hospital - Cornell Medical Center, New York, New York, USA.
Kräling
B M
BM
Stewart
R J
RJ
Chen
X
X
Bochner
B H
BH
Luster
A D
AD
Poppas
D P
DP
O'Donnell
M A
MA
D000970:Antineoplastic Agents / Q000494:pharmacology; D001500:BCG Vaccine / Q000494:pharmacology* / Q000627:therapeutic use; D002455:Cell Division; D054357:Chemokine CXCL10; D019743:Chemokines, CXC / Q000494:pharmacology; D016207:Cytokines / Q000502:physiology* / Q000652:urine; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008222:Lymphokines / Q000494:pharmacology; D018919:Neovascularization, Physiologic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014571:Urologic Neoplasms / Q000628:therapy; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000970:Antineoplastic Agents; D001500:BCG Vaccine; D054357:Chemokine CXCL10; D019743:Chemokines, CXC; D016207:Cytokines; D016228:Endothelial Growth Factors; D008222:Lymphokines; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D007371:Interferon-gamma