title: Tumor suppressive effects of WEE1 gene silencing could not enhance immunopotentiation effects of CD80 and 4-1BBL co-stimulation in human T cells.
abstract: BACKGROUND
  Activation of T cells against tumors by recruiting co-stimulatory molecules has been an attractive approach for cancer immunotherapy. Reports suggested that targeting different genes in tumors might also boost T cell-mediated tumor destruction.
  
  
  OBJECTIVE
  We investigated whether in vitro WEE1 gene silencing in MDA-MB-468 and MCF7 breast cancer cell lines could enhance immunopotentiating effects of CD80 and 4-1BBL co-stimulation in human T cells.
  
  
  METHODS
  WEE1 gene was specifically silenced in the cancer cells using shRNA technology. The co-stimulatory molecules were over-expressed on the surface of the cancer cells by recombinant non-replicative adenoviruses. The immune reaction of T cells in the co-culture with tumor cells was studied. IFN-g production was assessed by intracellular staining of T cells. To assess cytotoxic activity of CD8+ T cells, the CD107a mobilization-degranulation assay was performed. Expression of granzyme B, perforin and fasl were examined by real time PCR.
  
  
  RESULTS
  T cell dual co-stimulation led to a significant increase in the frequency of IFN-g producing cells and higher percentages of degranulation in CD8+ T cells. It also resulted in higher expression levels of the cytotoxicity-related genes. WEE1 gene silencing in the target cells alone however, could not produce significant immune reactivation in the cultured T cells. Likewise, the immune responses of T cells neither improved nor suppressed when dually co-stimulated PBMCs were exposed to the cancer cells with silenced WEE1.
  
  
  CONCLUSIONS
  In spite of antitumor effects of WEE1 silencing, combination of this approach with immune co-stimulation could not boost the reactivity of cultured T cells against the tested breast cancer cells.
authors.forename: Naghmeh
authors.initials: N
authors.lastname: Ghiasi
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Habibagahi
authors.forename: Rozita
authors.initials: R
authors.lastname: Rosli
authors.forename: Abbas
authors.initials: A
authors.lastname: Ghaderi
authors.forename: Khatijah
authors.initials: K
authors.lastname: Yusoff
authors.forename: Ahmad
authors.initials: A
authors.lastname: Hosseini
authors.forename: Syahril
authors.initials: S
authors.lastname: Abdullah
authors.affiliation: School of Medicine, Shiraz Institute for Cancer Research, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Mansooreh
authors.initials: M
authors.lastname: Jaberipour
mesh_terms: D053330:4-1BB Ligand / Q000494:pharmacology*; D018122:B7-1 Antigen / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D005260:Female; D020868:Gene Silencing; D064368:Healthy Volunteers; D006801:Humans; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D053330:4-1BB Ligand; D018122:B7-1 Antigen; D018797:Cell Cycle Proteins; D016207:Cytokines; D009687:Nuclear Proteins; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human