title: Transgene expression in various organs post BM-HSC transplantation.
abstract: Gene therapy mediated by bone marrow-derived hematopoietic stem cells (BM-HSC) has been widely used in treating genetic deficiencies in both pre-clinical and clinical settings. Using mitotically inactive cell-targeting lentivirus with separate promoters for our gene of interest (the murine MHC class II (MHCII) chaperone, invariant chain (Ii)) and a GFP reporter, we monitored the expression and function of introduced Ii in various types of professional antigen presenting cells (B cells, macrophages and DC) from different organs (spleen, pancreatic lymph nodes (PLN), BM and blood). Ii and GFP were detected. Ii levels correlated with GFP levels only in macrophages and monocytes from spleen, monocytes from PLN and macrophage precursors from blood. By cell type, Ii levels in PLN cells were more similar to those in spleen cells than to those in blood or BM cells. Functionally, Ii expressed in PLN or spleen had more effect on MHCII abundance than Ii expressed in BM or blood. The results have implications for analysis of the outcomes of gene therapy when both therapeutic and reporter genes are introduced. The findings also have implications for understanding the development of immune molecule function.
authors.affiliation: Department of Pediatrics, Program in Immunology, Program in Human Gene Therapy, Stanford University School of Medicine, Stanford, CA, USA. Electronic address: nanwang@stanford.edu.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: Department of Pediatrics, Program in Immunology, Program in Human Gene Therapy, Stanford University School of Medicine, Stanford, CA, USA. Electronic address: nrajas@stanford.edu.
authors.forename: Narendiran
authors.initials: N
authors.lastname: Rajasekaran
authors.affiliation: Department of Pediatrics, Program in Immunology, Program in Human Gene Therapy, Stanford University School of Medicine, Stanford, CA, USA. Electronic address: thou1@stanford.edu.
authors.forename: Tieying
authors.initials: T
authors.lastname: Hou
authors.affiliation: Department of Pediatrics, Program in Immunology, Program in Human Gene Therapy, Stanford University School of Medicine, Stanford, CA, USA. Electronic address: mellins@stanford.edu.
authors.forename: Elizabeth D
authors.initials: ED
authors.lastname: Mellins
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000166:cytology / Q000378:metabolism; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology*; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008198:Lymph Nodes / Q000378:metabolism; D051379:Mice; D011401:Promoter Regions, Genetic; D013154:Spleen / Q000378:metabolism; D019076:Transgenes / Q000235:genetics*
chemical_list: D000944:Antigens, Differentiation, B-Lymphocyte; D000949:Histocompatibility Antigens Class II; C045084:invariant chain; D049452:Green Fluorescent Proteins
keywords: AA; ADL; APC; Ab; BM; DC; EF1a; GFP; GM-CSF; HSC; IRES; Ii; Lin; MFI; MHCII; MOI; MPB; MSCV; PLN; T2A; Thoseaasigna virus 2A; adrenoleukodystrophy; amino acid; antibody; antigen presenting cells; bone marrow; dendritic cells; elongation factor 1a; granulocyte-macrophage colony-stimulating factor; green fluorescent protein; hematopoietic stem cells; internal ribosome entry sites; invariant chain; linage; major histocompatibility complex class II; mean fluorescence intensities; mobilized peripheral blood; multiplicity of infection; murine stem cell virus; pancreatic lymph nodes