title: Targeting rare populations of murine antigen-specific T lymphocytes by retroviral transduction for potential application in gene therapy for autoimmune disease.
abstract: CD4+ T cells are important mediators in the pathogenesis of autoimmunity and would therefore provide ideal candidates for lymphocyte-based gene therapy. However, the number of Ag-specific T cells in any single lesion of autoimmunity may be quite low. Successful gene transfer into autoantigen-specific CD4+ T cells would serve as an ideal vehicle for site-targeted gene therapy if it were possible to transduce preferentially the small number of autoantigen-specific T cells. In this study we have demonstrated that retroviral infection of CD4+ lymphocytes from either autoantigen-stimulated TCR transgenic mice, or Ag-activated immunized nontransgenic mice, with a retroviral vector (pGCIRES), resulted in the transduction of only the limited number of Ag-reactive CD4+ T cells. In contrast, polyclonal activation of the same cultures resulted in transduction of non-antigen-specific lymphocytes. Transduction of Ag-reactive CD4+ T cells with pGCIRES retrovirus encoding the regulatory genes IL-4 (IL4) and soluble TNF receptor (STNFR) resulted in stable integration and long-term expression of recombinant gene products. Moreover, expression of the pGCIRES marker protein, GFP, directly correlated with the expression of the upstream regulatory gene. Retroviral transduction of CD4+ T cells targeted specifically Ag-reactive cells and was cell cycle-dependent and evident only during the mitosis phase. These studies suggest that retroviral transduction of autoantigen-specific murine CD4+ T cells, using the pGCIRES retroviral vector, may provide a potential method to target and isolate the low frequency of autoantigen-specific murine CD4+ T cells, and provides a rational approach to gene therapy in animal models of autoimmunity.
authors.affiliation: Department of Medicine, Division of Immunology, Stanford University School of Medicine, Stanford CA 94305, USA. ginac@leland.stanford.edu
authors.forename: G L
authors.initials: GL
authors.lastname: Costa
authors.forename: J M
authors.initials: JM
authors.lastname: Benson
authors.forename: C M
authors.initials: CM
authors.lastname: Seroogy
authors.forename: P
authors.initials: P
authors.lastname: Achacoso
authors.forename: C G
authors.initials: CG
authors.lastname: Fathman
authors.forename: G P
authors.initials: GP
authors.lastname: Nolan
mesh_terms: D020413:3' Untranslated Regions / Q000276:immunology; D016475:3T3 Cells; D020121:5' Untranslated Regions / Q000276:immunology; D000818:Animals; D001324:Autoantigens / Q000276:immunology; D001327:Autoimmune Diseases / Q000235:genetics* / Q000276:immunology / Q000628:therapy*; D015704:CD4 Antigens / Q000096:biosynthesis; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D018984:Epitopes, T-Lymphocyte / Q000235:genetics*; D005786:Gene Expression Regulation / Q000276:immunology; D018390:Gene Targeting; D017930:Genes, Reporter / Q000276:immunology; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000138:chemical synthesis; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008213:Lymphocyte Activation / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D008938:Mitosis / Q000235:genetics / Q000276:immunology; D008979:Moloney murine leukemia virus / Q000235:genetics* / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000378:metabolism*; D014161:Transduction, Genetic / Q000276:immunology*
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D001324:Autoantigens; D015704:CD4 Antigens; D018984:Epitopes, T-Lymphocyte; D008164:Luminescent Proteins; D011948:Receptors, Antigen, T-Cell; D049452:Green Fluorescent Proteins