title: Phenotypic correction of primary Fanconi anemia T cells with retroviral vectors as a diagnostic tool.
abstract: OBJECTIVE
  The aim of this study was to develop a rapid laboratory procedure that is capable of subtyping Fanconi anemia (FA) complementation groups FA-A, FA-C, FA-G, and FA-nonACG patients from a small amount of peripheral blood.
  
  
  METHODS
  For this test, primary peripheral blood-derived FA T cells were transduced with oncoretroviral vectors that expressed FANCA, FANCC, or FANCG cDNA. We achieved a high efficiency of gene transfer into primary FA T cells by using the fibronectin fragment CH296 during transduction. Transduced cells were analyzed for correction of the characteristic DNA cross-linker hypersensitivity by cell survival or by metaphase analyses.
  
  
  RESULTS
  Retroviral vectors containing the cDNA for FA-A, FA-C, and FA-G, the most frequent complementation groups in North America, allowed rapid identification of the defective gene by complementation of primary T cells from 12 FA patients.
  
  
  CONCLUSIONS
  Phenotypic correction of FA T cells using retroviral vectors can be used successfully to determine the FA complementation group immediately after diagnosis of the disease.
authors.affiliation: Klinik für Pädiatrische Hämatologie und Onkologie, Zentrum für Kinderheilkunde, Heinrich-Heine-Universität, Düsseldorf, Germany. Helmut.Hanenberg@uni-duesseldorf.de
authors.forename: Helmut
authors.initials: H
authors.lastname: Hanenberg
authors.forename: Sat Dev
authors.initials: SD
authors.lastname: Batish
authors.forename: Karen E
authors.initials: KE
authors.lastname: Pollok
authors.forename: Lydia
authors.initials: L
authors.lastname: Vieten
authors.forename: Peter C
authors.initials: PC
authors.lastname: Verlander
authors.forename: Cordula
authors.initials: C
authors.lastname: Leurs
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Cooper
authors.forename: Kerstin
authors.initials: K
authors.lastname: Göttsche
authors.forename: Laura
authors.initials: L
authors.lastname: Haneline
authors.forename: D Wade
authors.initials: DW
authors.lastname: Clapp
authors.forename: Stephan
authors.initials: S
authors.lastname: Lobitz
authors.forename: David A
authors.initials: DA
authors.lastname: Williams
authors.forename: Arleen D
authors.initials: AD
authors.lastname: Auerbach
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002460:Cell Line; D002461:Cell Line, Transformed; D002470:Cell Survival; D004268:DNA-Binding Proteins; D005006:Ethnicity; D005199:Fanconi Anemia / Q000097:blood / Q000175:diagnosis / Q000235:genetics* / Q000276:immunology*; D052217:Fanconi Anemia Complementation Group A Protein; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006367:HeLa Cells; D004854:Herpesvirus 4, Human; D006801:Humans; D016130:Immunophenotyping; D008677:Metaphase; D051379:Mice; D016685:Mitomycin / Q000494:pharmacology; D011506:Proteins / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D012190:Retroviridae; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000473:pathology; D014162:Transfection
chemical_list: D004268:DNA-Binding Proteins; C498471:FANCA protein, human; C120277:Fanca protein, mouse; D052217:Fanconi Anemia Complementation Group A Protein; D011506:Proteins; D011994:Recombinant Proteins; D016685:Mitomycin