879 BLADDER EXPRESSION OF CYTOTOXIC EFFECTOR MOLECULES IN RESPONSE TO INTRAVESICAL MYCOBACTERIUM BOVIS BACILLUS CALMETTE-GUÉRIN (BCG)

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research1 Apr 2011879 BLADDER EXPRESSION OF CYTOTOXIC EFFECTOR MOLECULES IN RESPONSE TO INTRAVESICAL MYCOBACTERIUM BOVIS BACILLUS CALMETTE-GUÉRIN (BCG) Matthew Knudson, Jonathan Henning, Michael O'Donnell, and Yi Luo Matthew KnudsonMatthew Knudson Iowa City, IA More articles by this author , Jonathan HenningJonathan Henning Iowa City, IA More articles by this author , Michael O'DonnellMichael O'Donnell Iowa City, IA More articles by this author , and Yi LuoYi Luo Iowa City, IA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.703AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Bladder expression of cytokines and chemokines in response to intravesical (i.b.) BCG has been well documented. However, studies on BCG induction of bladder expression of cytotoxic effector molecules are underdeveloped. We investigated a panel of cytotoxic effector molecules, including IFN-γ, TNF-α, perforin, granzyme B, Fas ligand (FasL), and tumor necrosis factor related apoptosis-inducing ligand (TRAIL), in the BCG-treated bladders to determine whether expression of these molecules was correlated with the effect of BCG in treating bladder cancer. METHODS C57BL/6 female mice were treated i.b. with Pasteur strain BCG (0.1 OD/dose) twice weekly for a total of 7 treatments. Mice treated with PBS served as a control. Mice were sacrificed for bladder RNA extraction after treatments 2, 4, 6 and 7. RT-PCR was used to analyze mRNA levels of IFN-γ TNF-α, perforin, granzyme B, FasL and TRAIL. IP-10 (a Th1 chemokine) and TGF-β (an inhibitory cytokine) were included. GAPDH was used as an internal control. ImageJ software was used to quantify PCR products on agarose gels. In a parallel experiment C57BL/6 female mice were implanted i.b. with 5 X 105 syngeneic bladder cancer MB49 cells that expressed luciferase (MB49-Luc) at day 0 and treated i.b. with BCG (0.1 OD/dose) twice weekly starting at day 1 for a total of 6 treatments. Control mice were treated with PBS. Bioluminescence was measured once weekly with Xenogen IVIS. At day 23 mice were euthanized and bladder weights measured. Statistics was performed by one-way analysis of variance (ANOVA) of natural log transformed bladder weights followed by a post hoc Tukey's test. RESULTS I.b. BCG treatment led to an increase in bladder mRNAs of IFN-γ,TNF-α, IP-10 and perforin and a decrease in bladder mRNAs of TGF-β. There were no changes in bladder mRNAs of granzyme B, FasL and TRAIL. In MB49-Luc orthotopic tumor model, i.b. BCG treatment reduced bladder weight by 53% (p = 0.0503 vs. PBS-treated group). In addition, i.b. BCG treatment also led to a delay in tumor growth by bioluminescence compared to PBS-treated group. CONCLUSIONS Intravesical administration of BCG induced bladder expression of cytotoxic effector molecules IFN-γ, TNF-α and perforin, which was correlated with effective BCG treatment of bladder cancer. Increased expression of IP-10 and decreased expression of TGF-β, together with increased IFN-γ and TNF-α, support the role of BCG in the induction of Th1 immune responses in the bladders. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e352 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Matthew Knudson Iowa City, IA More articles by this author Jonathan Henning Iowa City, IA More articles by this author Michael O'Donnell Iowa City, IA More articles by this author Yi Luo Iowa City, IA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...