title: Murine β-defensin-2 may regulate the effect of bacillus Calmette-Guérin (BCG) in normal mouse bladder.
abstract: OBJECTIVE
  We investigated whether bacillus Calmette-Guérin (BCG)-induced secretion of murine β-defensin-2 (mBD2) and determined whether mBD2 regulated BCG effects in the normal mouse bladder.
  
  
  METHODS
  A total of 140 C57BL/6 female mice were divided into 28 groups, and the experiment was performed over 3 steps. In the first step (20 groups), mice bladders were stimulated with different doses of BCG (multiplicity of infection [MOI] 0, 1, 10, 30, and 100) and histological analysis was conducted in bladder specimens isolated at different times (0, 4, 8, and 24h after instillation) to determine optimal dose and time point of BCG internalization and urine mBD2 and cytokine concentration. In the second step (4 groups), BCG internalization and urine cytokine levels were measured after pretreatment of different recombinant mBD2 (rmBD2) (0, 1, 2.5, and 5 ng/ml) at optimal dose and time point. In the third step (4 groups), BCG internalization and urine cytokine levels were compared between pretreatment conditions (control, rmBD2, anti-mBD2 Ab, and rmBD2+anti-mBD2 Ab). Urine was collected for estimating mBD2 levels and a multiplex analysis for 9 cytokines. Real-time polymerase chain reaction assay was used for estimating the relative BCG cell number in mice bladder tissue.
  
  
  RESULTS
  Bladder edema was induced by BCG (MOI 30 and 100), which progressed to an inflammatory infiltrate composed primarily of neutrophils and increased mBD2 secretion at 4 hours after instillation. Relative BCG cell number and urinary cytokine levels (interferon-γ and interleukins [IL]-2, -4, -6, and -10) response pattern was characterized by a peak at 4 hours after instillation followed by rapid decline. The levels of interferon-γ, and IL-1β, -2, -4, -6, and -10 and relative BCG cell numbers decreased in a dose-dependent manner according to pretreatment with rmBD2 protein, and the responses were potentiated in the anti-mBD2 pretreatment group at 4 hours after BCG (MOI 30) instillation.
  
  
  CONCLUSIONS
  The present results suggest that the mouse urothelium produces mBD2 in response to intravesicular BCG as a defense mechanism against BCG, and blocking mBD2 by an anti-mBD2 antibody increased the effectiveness of BCG.
authors.affiliation: Department of Urology, Chung-Ang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Jong Kyou
authors.initials: JK
authors.lastname: Kwon
authors.affiliation: Department of Urology, Chung-Ang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Byung Hoon
authors.initials: BH
authors.lastname: Chi
authors.affiliation: Department of Urology, Chung-Ang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Se Young
authors.initials: SY
authors.lastname: Choi
authors.affiliation: Department of Urology, Chung-Ang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Soon-Ja
authors.initials: SJ
authors.lastname: Kim
authors.affiliation: Department of Pathology, Chung-Ang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Tae-Jin
authors.initials: TJ
authors.lastname: Lee
authors.affiliation: Department of Microbiology, Chung-Ang University College of Medicine, Seoul, Republic of Korea.
authors.forename: Kijeong
authors.initials: K
authors.lastname: Kim
authors.affiliation: Department of Urology, Chung-Ang University Hospital, Seoul, Republic of Korea. Electronic address: caucih@cau.ac.kr.
authors.forename: In Ho
authors.initials: IH
authors.lastname: Chang
mesh_terms: D000818:Animals; D001500:BCG Vaccine / Q000627:therapeutic use*; D016207:Cytokines / Q000378:metabolism; D004487:Edema / Q000473:pathology; D005260:Female; D005786:Gene Expression Regulation; D051379:Mice; D008810:Mice, Inbred C57BL; D009504:Neutrophils / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D001743:Urinary Bladder / Q000187:drug effects*; D019459:Urothelium / Q000378:metabolism; D023083:beta-Defensins / Q000378:metabolism*
chemical_list: D001500:BCG Vaccine; D016207:Cytokines; C117047:Defb2 protein, mouse; D011994:Recombinant Proteins; D023083:beta-Defensins
keywords: Bacillus Calmette-Guérin; Bladder; Murine β-defensin-2