title: Efficient lentiviral transduction of primary human acute myelogenous and lymphoblastic leukemia cells.
abstract: OBJECTIVE
  Gene manipulation and cell vaccines represent innovative strategies to enhance the immunogenicity of cancer cells. We adopted a defective lentivirus derived from the human immunodeficiency virus (HIV)-1 backbone and carrying the enhanced green fluorescent protein (EGFP) gene to transduce primary human acute myelogenous leukemia (AML) and B-precursor acute lymphoblastic leukemia (ALL) cells.
  
  
  METHODS
  AML blasts were maintained with or without cytokines (stem cell factor, FLT3 ligand and interleukin 3) for 48 hours, and successively infected with two spin infection cycles. ALL blasts were cultured on a murine S17 stromal cell line.
  
  
  RESULTS
  As regards AML cells, the efficiency of infection at 7 days varied from 8.4 to 37%. As confirmed by cell cycle analysis, cells were, in most of the cases, blocked in different phases of the cycle and did not proliferate during culture: the infection was therefore obtained in the absence of cell proliferation. In contrast, the maintenance of optimal cell viability was of fundamental importance for obtaining good infection levels. As regards ALL blasts, the percentages of infection after 3 days varied from 4.4 to 21%.
  
  
  CONCLUSIONS
  These preliminary data suggest that gene delivery into primary human AML and B-precursor ALL cells by an HIV-1 derived lentiviral vector could represent a strategy for engineering leukemic cells for use as cancer vaccines.
authors.affiliation: Centro M. Tettamanti, Clinica Pediatrica Università di Milano Bicocca, Ospedale San Gerardo, Monza, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Biagi
authors.forename: F
authors.initials: F
authors.lastname: Bambacioni
authors.forename: G
authors.initials: G
authors.lastname: Gaipa
authors.forename: C
authors.initials: C
authors.lastname: Casati
authors.forename: J
authors.initials: J
authors.lastname: Golay
authors.forename: A
authors.initials: A
authors.lastname: Biondi
authors.forename: M
authors.initials: M
authors.lastname: Introna
mesh_terms: D001854:Bone Marrow Cells / Q000378:metabolism / Q000473:pathology; D002453:Cell Cycle; D002455:Cell Division; D017930:Genes, Reporter; D005991:Glycerol Kinase / Q000235:genetics; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000473:pathology; D008164:Luminescent Proteins / Q000235:genetics; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics* / Q000473:pathology; D011401:Promoter Regions, Genetic; D014161:Transduction, Genetic / Q000379:methods* / Q000592:standards; D014407:Tumor Cells, Cultured
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D005991:Glycerol Kinase