Lentiviral vectors pseudotyped with glycoproteins from ross river and vesicular stomatitis viruses

Variable transduction related to cell type and culture conditions

Christoph Kahl, Karen Pollok, Laura S. Haneline, Kenneth Cornetta

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

HIV-1-derived lentiviral vectors have been pseudotyped with various envelope glycoproteins to alter their host range. Previously, we found that envelope glycoproteins derived from the alphavirus Ross River virus (RRV) can pseudotype lentiviral vectors and mediate efficient transduction of a variety of epithelial and fibroblast-derived cell lines. In this study, we have investigated transduction of hematopoietic cells using RRV-pseudotyped vectors encoding the enhanced green fluorescent protein (EGFP). RRV-mediated transduction of human CD34+ cord blood cells and progenitors was very inefficient, even at multiplicities of infection of 100 (0.4% EGFP-positive progenitor colonies). Inefficient transduction was also observed in a variety of hematopoietic cell lines. However, two erythroleukemia-derived cell lines and monocytic cells that were driven to macrophage-like differentiation were moderately transduced. Transduction of hematopoietic cells with a control VSV-G-pseudotyped lentiviral vector was generally efficient, but unexpectedly decreased up to threefold upon stimulation of lymphocytic cell lines or primary murine bone marrow cells. Also, the tested hematopoietic cell lines were essentially nonpermissive for adeno-associated type 2 (AAV) vectors, and this was not affected by lineage, activity, or differentiation. Treatment of permissive 293 cells with proteases revealed that transduction with both the RRV- and the VSV-G-pseudotyped vectors in part depends on the presence of cell surface proteins. These results show a severely restricted ability of RRV glycoproteins to mediate transduction in hematopoietic cells that is likely due to specific receptor requirements that differ from those of VSV-G and AAV. Conversely, transduction with the VSV glycoprotein is affected by cellular activation more than widely believed. Our findings suggest that the envelope glycoproteins and culture conditions employed need to be carefully evaluated for each application. Furthermore, the uniquely restricted host range of RRV-pseudotyped vectors may aid in the design of novel cell-selective transduction strategies.

Original languageEnglish (US)
Pages (from-to)470-482
Number of pages13
JournalMolecular Therapy
Volume11
Issue number3
DOIs
StatePublished - Mar 2005
Externally publishedYes

Fingerprint

Ross River virus
Vesicular Stomatitis
Rivers
Glycoproteins
Cell Culture Techniques
Viruses
Cell Line
Host Specificity
Alphavirus
Leukemia, Erythroblastic, Acute
Fetal Blood
Bone Marrow Cells
HIV-1
Blood Cells
Membrane Proteins
Peptide Hydrolases
Fibroblasts
Macrophages
Infection

Keywords

  • Lentiviral vectors
  • Pseudotyping
  • Ross River virus
  • VSV-G

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Lentiviral vectors pseudotyped with glycoproteins from ross river and vesicular stomatitis viruses : Variable transduction related to cell type and culture conditions. / Kahl, Christoph; Pollok, Karen; Haneline, Laura S.; Cornetta, Kenneth.

In: Molecular Therapy, Vol. 11, No. 3, 03.2005, p. 470-482.

Research output: Contribution to journalArticle

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