title: [Immobilization of streptavidin-tagged bioactive hTNF-alpha on biotinylated mucosal surface of the bladder wall for treatment of superficial bladder cancer in mice].
abstract: OBJECTIVE
  To investigate a novel immunotherapy through immobilization of streptavidin-tagged hTNF-alpha on the biotinylated mucosal surface of the bladder wall for bladder cancer treatment in mice.
  
  
  METHODS
  A total of 120 female C57BL/6j mice were randomized into 5 equal groups, namely blank control, PBS, soluble hTNF-alpha, SA-GFP, and SA-hTNF-alpha treatment groups. Twenty-four hours after establishment of a mouse model of orthotopic superficial bladder cancer, SA-hTNF-alpha fusion protein was immobilized on the biotinylated mucosal surface of the bladder wall, which was repeated every 4 days for a total of 6 sessions. Immunohistochemistry was performed to detect the retention time of SA-hTNF-alpha fusion protein in the biotinylated mouse bladder mucosa and the distribution of CD4(+) and CD8(+) lymphocytes in the mucosa and tumor tissues, with the tumor growth and mouse survival also observed. The cytotoxiciy of the tumor-specific lymphocytes was evaluated. The mice responding well to the treatment were re-challenged by MB49 and monitored for survival.
  
  
  RESULTS
  SA-hTNF-alpha could be efficiently and stably immobilized on the bladder mucosal surface for as long as 7 days. On day 60 after MB49 implantation, 18 out of 22 SA- hTNF-alpha-treated mice survived, with 9 appearing tumor-free, but all the mice in PBS control group died. Five out of 9 tumor-free mice in SA-hTNF-alpha group showed resistance to a re-challenge with intravesical MB49. The numbers of CD4(+) and CD8(+) lymphocytes were significantly greater in SA-hTNF-alpha group than in the other groups (P<0.05). The cytotoxicity of the tumor-specific lymphocytes was significantly stronger in SA-hTNF-alpha group than in the other groups (P<0.05).
  
  
  CONCLUSIONS
  SA-hTNF-alpha immobilized on the biotinylated mucosal surface of the bladder wall can significantly inhibit the tumor growth and promote the survival of the mice bearing orthotopic superficial bladder cancer.
authors.affiliation: Department of Urologic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. chzh@ymail.com
authors.forename: Zhong
authors.initials: Z
authors.lastname: Chen
authors.forename: Wan-long
authors.initials: WL
authors.lastname: Tan
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.forename: Zhong-kun
authors.initials: ZK
authors.lastname: Liang
authors.forename: Cui-xiang
authors.initials: CX
authors.lastname: Xu
authors.forename: Ji-min
authors.initials: JM
authors.lastname: Gao
mesh_terms: D000283:Administration, Intravesical; D000818:Animals; D019921:Biotinylation; D002295:Carcinoma, Transitional Cell / Q000276:immunology / Q000628:therapy; D005260:Female; D055767:Immobilized Proteins / Q000627:therapeutic use*; D007167:Immunotherapy / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000627:therapeutic use; D019809:Streptavidin / Q000378:metabolism / Q000627:therapeutic use*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism / Q000627:therapeutic use*; D001749:Urinary Bladder Neoplasms / Q000276:immunology / Q000628:therapy*
chemical_list: D055767:Immobilized Proteins; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D019809:Streptavidin