Volume 13, Issue 1, Pages 42-48 (January 2006) Bone marrow transplantation combined with gene therapy to induce antigen-specific tolerance and ameliorate EAE  Biying Xu, Peter Haviernik, Lawrence A. Wolfraim, Kevin D. Bunting, David W. Scott  Molecular Therapy  Volume 13, Issue 1, Pages 42-48 (January 2006) DOI: 10.1016/j.ymthe.2005.09.002 Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

FIG. 1 Expression of GFP in bone marrow cells transduced with MSCV-irGFP constructs. (A and B) Expression of GFP in hematopoietic stem cells and their descendants. (A) Flow cytometry for GFP expression after transduction and culture in vitro. (B) Expression in bone marrow, spleen, and peripheral blood at 8 weeks after BMT. (C) Expression of GFP+ cells in peripheral blood is stable. Persistence of expression in vivo at 10 and 14 weeks (PBL). (D) GFP expression in BMT mice is not lineage biased. Expression in different hematopoietic lineages (at 15 weeks after reconstitution). Molecular Therapy 2006 13, 42-48DOI: (10.1016/j.ymthe.2005.09.002) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

FIG. 2 T cell proliferation is decreased in recipients of PLPfl BM. Splenic T cell proliferation from mice given BM expressing PLPfl construct versus T cells from mice given control vector alone on day 15 after immunization. See Refs. [12] and [14] for details. Molecular Therapy 2006 13, 42-48DOI: (10.1016/j.ymthe.2005.09.002) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

FIG. 3 Effect of gene therapy on EAE induction. Mice receiving BM with PLPfl construct were completely protected from EAE, whereas mice receiving OVA peptide-transduced BM cells developed a typical relapsing–remitting clinical course. This is one of three separate, reproducible experiments. Molecular Therapy 2006 13, 42-48DOI: (10.1016/j.ymthe.2005.09.002) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

FIG. 4 Histology of brain sections in recipients of BM transduced with PLPfl (experimental, left) versus control vector (right). Massive perivascular infiltration (arrows) is seen in the brains of recipients of control vector-transduced BM but not in recipients of PLPfl BM. Hematoxylin and eosin staining. Molecular Therapy 2006 13, 42-48DOI: (10.1016/j.ymthe.2005.09.002) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

FIG. 5 Reduction in symptoms in mice that received BMT on day 12 postimmunization with PLP139-151 in CFA. Groups were randomized, irradiated with 900 rad, and given either PLPfl-transduced BM or control (GFP) vector-transduced BM. A significantly lower disease score was observed in mice receiving PLPfl for 60 days, with no relapse. Two separate experiments are shown. Molecular Therapy 2006 13, 42-48DOI: (10.1016/j.ymthe.2005.09.002) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

FIG. 6 Modulation of relapsing EAE with nonmyeloablative irradiation and PLPfl BM gene therapy. Mice were immunized with PLP peptide and adjuvant to induce disease and then given 300 rad plus transduced BM on day 6, 12, or 22. Three independent experiments are shown. Molecular Therapy 2006 13, 42-48DOI: (10.1016/j.ymthe.2005.09.002) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

FIG. 7 Structure of bicistronic vectors used in this study. Molecular Therapy 2006 13, 42-48DOI: (10.1016/j.ymthe.2005.09.002) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions