Characterization of an MDR1 retroviral bicistronic vector for correction of X-linked severe combined immunodeficiency

S. E. Kleiman, I. Pastan, J. M. Puck, M. M. Gottesman*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

X-linked severe combined immunodeficiency (XSCID) is a hereditary disorder characterized by sever T cell lymphopenia and abnormal B cell function. The disease is caused by mutations in IL2RG, the gene encoding the interleukin-2 receptor common γ chain (γc) shared by several interleukin receptors. A Harvey retroviral bicistronic vector containing an IL2RG cDNA and cDNA encoding the multidrug transporter (MDR1) was constructed to investigate the correction of XSCID. Tranlation of the MDR1 cDNA is achieved from an internal ribosome entry site (IRES). Mouse fibroblasts transfected or transduced with the vector expressed both membrane proteins as detected with specific monoclonal antibodies by fluorescence activated cell sorting. Two human XSCID B cell lines were transduced using a filter concentration method in combination with phosphate depletion. Significant expression of both proteins was detected by Western blot analysis. This construct might be particularly useful if high expression of γc is required as might be achievable through in vivo selection for drug resistance of recipient lymphocytes.

Original languageEnglish
Pages (from-to)671-676
Number of pages6
JournalGene Therapy
Volume5
Issue number5
DOIs
StatePublished - 1998
Externally publishedYes

Keywords

  • Gene transfer
  • MDR1
  • Retroviral vectors
  • XSCID

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