Fibronectin improves transduction of reconstituting hematopoietic stem cells by retroviral vectors: evidence of direct viral binding to chymotryptic carboxy-terminal fragments

Blood. 1996 Aug 1;88(3):855-62.


Efficient transduction of reconstituting hematopoletic stem cells (HSC) is currently only possible by cocultivation of target cells directly on producer cell lines, a method not applicable to human gene therapy protocols. Our laboratory has previously shown adhesion of primitive hematopoletic stem and progenitor cells to the carboxy-terminal 30/35-kD fragment of the extracellular matrix molecule fibronectin (FN 30/35) (Nature 352:438, 1991) and increased transduction of human hematopoietic progenitor cells via retroviral vectors while adherent to this fragment (J Clin Invest 93:1451, 1994). Here we report that (1) transduction of reconstituting murine HSC assayed 12 months after infection with retrovirus supernatant on FN 30/35 is as effective as cocultivation directly on producer cells; (2) recombinant retrovirus particles directly adhere to FN 30/35 in a quantitative and dose-dependent fashion; and (3) increased transduction efficiency on FN 30/ 35 does not appear to be associated with increased cell proliferation or activation of protein phosphorylation typically induced by integrin-fibronectin interactions. Therefore, we speculate that supernatant infection of HSC on FN 30/35 leads to colocalization of retrovirus particles and target cells on FN 30/35 molecule with a large increase in local virus titer presented to the cell. These findings have direct and important implications for the modification of current human gene therapy protocols.

Publication types

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

MeSH terms

  • 3T3 Cells
  • Adenosine Deaminase / genetics
  • Adenosine Deaminase / metabolism
  • Animals
  • Cell Cycle
  • Cells, Cultured
  • Chymotrypsin / metabolism
  • Fibronectins / metabolism
  • Fibronectins / pharmacology*
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism*
  • Hematopoietic Stem Cell Transplantation / methods*
  • Hematopoietic Stem Cells / drug effects*
  • Hematopoietic Stem Cells / metabolism
  • Hematopoietic Stem Cells / virology
  • Mice
  • Peptide Fragments / metabolism*
  • Recombinant Proteins / metabolism
  • Retroviridae / genetics
  • Retroviridae / metabolism*
  • Transfection / drug effects*


  • Fibronectins
  • Peptide Fragments
  • Recombinant Proteins
  • Chymotrypsin
  • Adenosine Deaminase