title: Safety and efficacy of activated transfected killer cells for neutropenic fungal infections.
abstract: BACKGROUND
  Invasive fungal infections cause considerable morbidity and mortality in neutropenic patients. White blood cell transfusions are a promising treatment for such infections, but technical barriers have prevented their widespread use.
  
  
  METHODS
  To recapitulate white blood cell transfusions, we are developing a cell-based immunotherapy using a phagocytic cell line, HL-60. We sought to stably transfect HL-60 cells with a suicide trap (herpes simplex virus thymidine kinase), to enable purging of the cells when desired, and a bioluminescence marker, to track the cells in vivo in mice.
  
  
  RESULTS
  Transfection was stable despite 20 months of continuous culture or storage in liquid nitrogen. Activation of these transfected cells with retinoic acid and dimethyl sulfamethoxazole enhanced their microbicidal effects. Activated transfected killer (ATAK) cells were completely eliminated after exposure to ganciclovir, confirming function of the suicide trap. ATAK cells improved the survival of neutropenic mice with lethal disseminated candidiasis and inhalational aspergillosis. Bioluminescence and histopathologic analysis confirmed that the cells were purged from surviving mice after ganciclovir treatment. Comprehensive necropsy, histopathology, and metabolomic analysis revealed no toxicity of the cells.
  
  
  CONCLUSIONS
  These results lay the groundwork for continued translational development of this promising, novel technology for the treatment of refractory infections in neutropenic hosts.
authors.affiliation: Division of Infectious Diseases, Los Angeles Biomedical Research Institute, Harbor-University of California at Los Angeles (UCLA) Medical Center, 1124 West Carson Street, Torrance, CA 90502, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.forename: Ashraf S
authors.initials: AS
authors.lastname: Ibrahim
authors.forename: Beverlie
authors.initials: B
authors.lastname: Baquir
authors.forename: Yue
authors.initials: Y
authors.lastname: Fu
authors.forename: David
authors.initials: D
authors.lastname: Applebaum
authors.forename: Julie
authors.initials: J
authors.lastname: Schwartz
authors.forename: Amy
authors.initials: A
authors.lastname: Wang
authors.forename: Valentina
authors.initials: V
authors.lastname: Avanesian
authors.forename: Brad
authors.initials: B
authors.lastname: Spellberg
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D001228:Aspergillosis / Q000150:complications / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D002177:Candidiasis / Q000150:complications / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D005260:Female; D007694:Killer Cells, Natural / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D009503:Neutropenia / Q000628:therapy*; D016019:Survival Analysis; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
chemical_list: D013937:Thymidine Kinase
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