title: Characterization of an MDR1 retroviral bicistronic vector for correction of X-linked severe combined immunodeficiency.
abstract: X-linked severe combined immunodeficiency (XSCID) is a hereditary disorder characterized by severe T cell lymphopenia and abnormal B cell function. The disease is caused by mutations in IL2RG, the gene encoding the interleukin-2 receptor common gamma chain (gamma c) 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. Translation 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 gamma c is required, as might be achievable through in vivo selection for drug resistance of recipient lymphocytes.
authors.affiliation: Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4255, USA.
authors.forename: S E
authors.initials: SE
authors.lastname: Kleiman
authors.forename: I
authors.initials: I
authors.lastname: Pastan
authors.forename: J M
authors.initials: JM
authors.lastname: Puck
authors.forename: M M
authors.initials: MM
authors.lastname: Gottesman
mesh_terms: D016475:3T3 Cells; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000818:Animals; D002461:Cell Line, Transformed; D002469:Cell Separation; D018432:Drug Resistance, Multiple; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D019450:Genes, MDR; D005819:Genetic Markers; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D009848:Olivomycins / Q000494:pharmacology; D015375:Receptors, Interleukin-2 / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D016511:Severe Combined Immunodeficiency / Q000628:therapy*; D014162:Transfection; D014750:Vincristine / Q000494:pharmacology
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D005819:Genetic Markers; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D009848:Olivomycins; D014750:Vincristine