title: Adeno associated viral-mediated intraosseous labeling of bone marrow derived cells for CNS tracking.
abstract: Inflammation, including microglial activation in the CNS, is an important hallmark in many neurodegenerative diseases. Microglial stimuli not only impact the brain microenvironment by production and release of cytokines and chemokines, but also influence the activity of bone marrow derived cells and blood born macrophage populations. In many diseases including brain disorders and spinal cord injury, researchers have tried to harbor the neuroprotective and repair properties of these subpopulations. Hematopoietic bone marrow derived cells (BMDCs) are of great interest, especially during gene therapy because certain hematopoietic cell subpopulations traffic to the sites of injury and inflammation. The aim of this study was to develop a method of labeling endogenous bone marrow derived cells through intraosseous impregnation of recombinant adeno-associated virus (rAAV) or lentivirus. We utilized rAAV serotype 9 (rAAV-9) or lentivirus for gene delivery of green florescence protein (GFP) to the mouse bone marrow cells. Flow cytometry showed that both viruses were able to efficiently transduce mouse bone marrow cells in vivo. However, the rAAV9-GFP viral construct transduced BMDCs more efficiently than the lentivirus (11.2% vs. 6.8%), as indicated by cellular GFP expression. We also demonstrate that GFP labeled cells correspond to bone marrow cells of myeloid origin using CD11b as a marker. Additionally, we characterized the ability of bone marrow derived, GFP labeled cells to extravasate into the brain parenchyma upon acute and subchronic neuroinflammatory stimuli in the mouse CNS. Viral mediated over expression of chemokine (C-C motif) ligand 2 (CCL2) or intracranial injection of lipopolysaccharide (LPS) recruited GFP labeled BMDCs from the periphery into the brain parenchyma compared to vehicle treated mice. Altogether our findings demonstrate a useful method of labeling endogenous BMDCs via viral transduction and the ability to track subpopulations throughout the body following insult or injury. Alternatively, this method might find utility in delivering therapeutic genes for neuroinflammatory conditions.
authors.affiliation: Dept of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.
authors.forename: Maj-Linda B
authors.initials: ML
authors.lastname: Selenica
authors.affiliation: Dept of Molecular Pharmacology & Physiology, Byrd Alzheimer's Institute, University of South Florida, Tampa, FL, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Reid
authors.affiliation: Dept of Molecular Pharmacology & Physiology, Byrd Alzheimer's Institute, University of South Florida, Tampa, FL, USA.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Pena
authors.affiliation: Dept of Molecular Pharmacology & Physiology, Byrd Alzheimer's Institute, University of South Florida, Tampa, FL, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Alvarez
authors.affiliation: Dept of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.
authors.forename: Jerry B
authors.initials: JB
authors.lastname: Hunt
authors.affiliation: Dept of Molecular Pharmacology & Physiology, Byrd Alzheimer's Institute, University of South Florida, Tampa, FL, USA.
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Nash
authors.affiliation: Dept of Molecular Pharmacology & Physiology, Byrd Alzheimer's Institute, University of South Florida, Tampa, FL, USA.
authors.forename: Dave
authors.initials: D
authors.lastname: Morgan
authors.affiliation: Dept of Molecular Pharmacology & Physiology, Byrd Alzheimer's Institute, University of South Florida, Tampa, FL, USA.
authors.forename: Marcia N
authors.initials: MN
authors.lastname: Gordon
authors.affiliation: Dept of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA. Electronic address: dlee1@health.usf.edu.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Lee
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000378:metabolism; D016026:Bone Marrow Transplantation; D001921:Brain / Q000378:metabolism / Q000473:pathology*; D039481:CD11b Antigen / Q000378:metabolism; D058948:Cell Tracking / Q000379:methods*; D000229:Dependovirus / Q000235:genetics*; D004195:Disease Models, Animal; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D007249:Inflammation / Q000235:genetics / Q000378:metabolism / Q000473:pathology* / Q000628:therapy; D016086:Lentivirus / Q000235:genetics; D008810:Mice, Inbred C57BL; D010641:Phenotype; D014161:Transduction, Genetic
chemical_list: D015415:Biomarkers; D039481:CD11b Antigen; D049452:Green Fluorescent Proteins
keywords: Bone marrow infection; GFP labeling; Infiltration; Inflammation; Monocyte; Recombinant adeno-associated virus serotype 9