Transduction of Schistosoma mansoni by vesicular stomatitis virus glycoprotein-pseudotyped Moloney murine leukemia retrovirus

Kristine J. Kines, Victoria H. Mann, Maria E. Morales, Bryan D. Shelby, Bernd H. Kalinna, Geoffrey N. Gobert, Sharon R. Chirgwin, Paul J. Brindley*

*Corresponding author for this work

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Retroviral transduction of cultured schistosomes offers a potential means to establish transgenic lines of schistosomes and thereby to facilitate the elucidation of schistosome gene function and expression. The Moloney murine leukemia retroviral (MMLV) vector pLNHX was modified to incorporate EGFP or luciferase reporter genes under control of schistosome endogenous gene promoters from the spliced leader RNA and HSP70 genes. These constructs and a plasmid encoding vesicular stomatitis virus glycoprotein (VSVG) were utilized along with GP2-293 cells to produce replication incompetent retrovirus particles pseudotyped with the VSVG envelope. Exposure of several developmental stages, including sporocysts, of Schistosoma mansoni to these virions was facilitated by incubation with polybrene and/or by centrifugation. The early stages of binding and uptake of virus to the parasite tegument were demonstrated by the immunofluorescence colocalization of VSVG envelope and retroviral capsid proteins. Southern hybridization analysis indicated the integration of proviral forms of the MMLV constructs in genomic DNA isolated from the virus exposed schistosomes. Furthermore, analysis of RNA isolated from virus treated parasites demonstrated the presence of transcripts encoding reporter transgenes. Together these results indicated productive transduction by VSVG pseudotyped MMLV of cultured schistosomes, and suggest a tractable route forward towards heritable schistosome transgenesis.

Original languageEnglish
Pages (from-to)209-220
Number of pages12
JournalExperimental Parasitology
Volume112
Issue number4
DOIs
Publication statusPublished - 01 Apr 2006
Externally publishedYes

Keywords

  • Moloney murine leukemia virus
  • Retrovirus
  • Schistosome
  • Transduction
  • Transgenesis
  • Vesicular stomatitis virus glycoprotein

ASJC Scopus subject areas

  • Parasitology
  • Immunology

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