title: Physiological relevance of LL-37 induced bladder inflammation and mast cells.
abstract: OBJECTIVE
  We established the physiological relevance of LL-37 induced bladder inflammation. We hypothesized that 1) human urinary LL-37 is increased in pediatric patients with spina bifida, 2) LL-37 induced inflammation occurs in our mouse model via urothelial binding and is dose dependent and 3) LL-37 induced inflammation involves mast cells.
  
  
  METHODS
  To test our first hypothesis, we obtained urine samples from 56 pediatric patients with spina bifida and 22 normal patients. LL-37 was measured by enzyme-linked immunosorbent assay. Our second hypothesis was tested in C57Bl/6 mice challenged with 7 LL-37 concentrations intravesically for 1 hour. At 24 hours tissues were examined histologically and myeloperoxidase assay was done to quantitate inflammation. In separate experiments fluorescent LL-37 was instilled and tissues were obtained immediately (time = 0) and at 24 hours (time = 24). To test our final hypothesis, we performed immunohistochemistry for mast cell tryptase and evaluated 5 high power fields per bladder to determine the mean number of mast cells per mm(2).
  
  
  RESULTS
  Urinary LL-37 was 89-fold higher in patients with spina bifida. Mouse LL-37 dose escalation experiments revealed increased inflammation at higher LL-37 concentrations. Fluorescent LL-37 demonstrated global urothelial binding at time = 0 but was not visible at time = 24. Immunohistochemistry for tryptase revealed mast cell infiltration in all tissue layers. At higher concentrations the LL-37 challenge led to significantly greater mast cell infiltration.
  
  
  CONCLUSIONS
  Urinary LL-37 was significantly increased in pediatric patients with spina bifida. To our knowledge we report for the first time that LL-37 can elicit profound, dose dependent bladder inflammation involving the urothelium. Finally, inflammation propagation involves mast cells.
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Siam
authors: S
authors: Oottamasathien
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Wanjian
authors: W
authors: Jia
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Lindsi McCoard
authors: LM
authors: Roundy
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Jianxing
authors: J
authors: Zhang
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Li
authors: L
authors: Wang
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Xiangyang
authors: X
authors: Ye
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: A Cameron
authors: AC
authors: Hill
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Justin
authors: J
authors: Savage
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Wong Yong
authors: WY
authors: Lee
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Ann Marie
authors: AM
authors: Hannon
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Sylvia
authors: S
authors: Milner
authors: Division of Pediatric Urology (SO, WJ, LMR, LW, ACH, AMH, SM) and Departments of Surgery (SO, WJ, LMR, LW, ACH, AMH, SM), Medicinal Chemistry (JZ, GDP) and Pharmacotherapy (XY) and Center for Therapeutic Biomaterials (JZ, GDP), University of Utah/Primary Children's Medical Center and GlycoMira Therapeutics, L.L.C. (JS, WYL), Salt Lake City, Utah.
authors: Glenn D
authors: GD
authors: Prestwich
mesh_terms: D000293:Adolescent; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000378:metabolism* / Q000633:toxicity; D002452:Cell Count; D002648:Child; D002675:Child, Preschool; D003556:Cystitis / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D007231:Infant, Newborn; D008070:Lipopolysaccharides; D008297:Male; D008407:Mast Cells / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D016135:Spinal Dysraphism / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D019459:Urothelium / Q000378:metabolism / Q000473:pathology; D054804:Cathelicidins
chemical_list: D023181:Antimicrobial Cationic Peptides; D008070:Lipopolysaccharides; D054804:Cathelicidins
keywords: DAB; GAG; IC/PBS; MM/SB; MPO; TBS; TBS-Tween®; TBST; UTI; cystitis; diaminobenzidine; glycosaminoglycan; inflammation; interstitial; interstitial cystitis/painful bladder syndrome; mast cells; meningomyelocele; myelomeningocele/spina bifida; myeloperoxidase; t; time; tris buffered saline; urinary bladder; urinary tract infection