Phenotypic correction of primary Fanconi anemia T cells with retroviral vectors as a diagnostic tool

Exp Hematol. 2002 May;30(5):410-20. doi: 10.1016/s0301-472x(02)00782-8.


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.

Materials and 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.

Conclusion: 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Animals
  • B-Lymphocytes / immunology
  • Cell Line
  • Cell Line, Transformed
  • Cell Survival
  • DNA-Binding Proteins*
  • Ethnicity
  • Fanconi Anemia / blood
  • Fanconi Anemia / diagnosis
  • Fanconi Anemia / genetics*
  • Fanconi Anemia / immunology*
  • Fanconi Anemia Complementation Group A Protein
  • Genetic Complementation Test
  • Genetic Vectors
  • HeLa Cells
  • Herpesvirus 4, Human
  • Humans
  • Immunophenotyping
  • Metaphase
  • Mice
  • Mitomycin / pharmacology
  • Proteins / genetics*
  • Recombinant Proteins / metabolism
  • Retroviridae
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / pathology
  • Transfection


  • DNA-Binding Proteins
  • FANCA protein, human
  • Fanca protein, mouse
  • Fanconi Anemia Complementation Group A Protein
  • Proteins
  • Recombinant Proteins
  • Mitomycin