Newly-developed Sendai virus vector for retinal gene transfer: reduction of innate immune response via deletion of all envelope-related genes

J Gene Med. 2008 Feb;10(2):165-76. doi: 10.1002/jgm.1142.

Abstract

Background: Recombinant Sendai virus vectors (rSeV) constitute a new class of cytoplasmic RNA vectors that have shown efficient gene transfer in various organs, including retinal tissue; however, the related immune responses remain to be overcome in view of clinical applications. We recently developed a novel rSeV from which all envelope-related genes were deleted (rSeV/dFdMdHN) and, in the present study, assess host immune responses following retinal gene transfer.

Methods: rSeV/dFdMdHN or conventional F-gene deleted rSeV (rSeV/dF) was injected into subretinal space of adult Wistar rats or C57BL/6 mice. The transgene expression and histopathological findings were assessed at various time points. Immunological assessments, including the expression of proinflammatory cytokines, natural killer (NK)-cell activity, as well as SeV-specific cytotoxic T lymphocytes (CTLs) and antibodies, were performed following vector injection.

Results: rSeV/dFdMdHN showed high gene transfer efficiency into the retinal pigment epithelium at an equivalent level to that seen with rSeV/dF. In the early phase, the upregulation of proinflammatory cytokines, local inflammatory cell infiltration and tissue damage that were all prominently seen in rSeV/dF injection were dramatically diminished using rSeV/dFdMdHN. NK cell activity was also decreased, indicating a reduction of the innate immune response. In the later phase, on the other hand, CTL activity and anti-SeV antibodies were similarly induced, even using rSeV/dFdMdHN, and resulted in transient transgene expression in both vector types.

Conclusions: Deletion of envelope-related genes of rSeV dramatically reduces the vector-induced retinal damage and may extend the utility for ocular gene transfer; however, further studies regulating the acquired immune response are required to achieve long-term transgene expression of rSeV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Death
  • Cytokines / metabolism
  • Cytopathogenic Effect, Viral
  • Gene Deletion*
  • Genetic Vectors / genetics*
  • Green Fluorescent Proteins / metabolism
  • Immunity, Innate / immunology*
  • Inflammation Mediators / metabolism
  • Killer Cells, Natural / immunology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Rats
  • Rats, Wistar
  • Respirovirus Infections
  • Retina / cytology
  • Retina / metabolism*
  • Retina / virology
  • Sendai virus / genetics*
  • Time Factors
  • Transduction, Genetic / methods*
  • Transgenes
  • Viral Envelope Proteins / genetics*

Substances

  • Cytokines
  • Inflammation Mediators
  • Viral Envelope Proteins
  • Green Fluorescent Proteins