The major goal of this preclinical IPCP project is the development of lentiviral vectors for gene therapy of HIV diseases. Project 1 focuses on developing HIV-based vectors that target hematopoietic cells, including stem cells and dendritic cells for antiviral and immune-based therapies.
Our specific aims are: 1. To establish optimal HIV-2 gene transfer vector and stable packaging lines-Efforts will be made to minimize the chance of RCR generation, while optimizing vector titers. 2. To demonstrate protection of primary T cells, macrophages and progeny of CD34+ cells by lentiviral vector transduction against HIV challenge-HIV-2 vectors expressing anti-HIV-1 and anti-CCR5 ribozymes will be tested in these studies. We will also ascertain whether transduction by these vectors will have any deleterious effect on cell proliferation, differentiation and/or function of these target cells. 3. To examine the effects of lentiviral vector transduction on dendritic cell function and interaction with T cells- Dendritic cells (DC) are both facilitators and defenders of HIV infection since they can present both viral antigens and infectious virus to T cells. Recent data suggest that while only immature DC's are infectable by HIV, mature DC's need to express functional chemokine receptors CCR5 and CXCR4 t transmit M-tropic and T-tropic viruses to CD4+ T cells respectively. We will construct HIV-1 vectors expressing gag-pol antigens as well as anti-CC4-5 and/or anti-CXCR4 ribozymes, and determine how transduction by these vectors will impact on the ability of dendritic cells to present antigen to CD4 and CD8 cells or to transmit HIV infection to CD4 cells. In addition to these aims, this project will collaborate closely with Projects 2 and 3 to directly compare the safety and efficacy of HIV-based and FIV-based vectors and MLV-based retroviral vectors in vitro and in vivo.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI045992-04
Application #
6642250
Study Section
Special Emphasis Panel (ZAI1)
Project Start
2002-08-01
Project End
2003-07-31
Budget Start
Budget End
Support Year
4
Fiscal Year
2002
Total Cost
$247,465
Indirect Cost
Name
University of California San Diego
Department
Type
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Cheng, Lingyun; Toyoguchi, Mitsuko; Looney, David J et al. (2005) Efficient gene transfer to retinal pigment epithelium cells with long-term expression. Retina 25:193-201
Morris, Kevin V; Looney, David J (2005) Characterization of human immunodeficiency virus (HIV)-2 vector mobilization by HIV-1. Hum Gene Ther 16:1463-72
Morris, Kevin V; Chan, Simon W-L; Jacobsen, Steven E et al. (2004) Small interfering RNA-induced transcriptional gene silencing in human cells. Science 305:1289-92
Gruber, A; Chen, I; Kuhen, K L et al. (2003) Generation of dendritic cells from lentiviral vector-transduced CD34+ cells from HIV+ donors. J Med Virol 70:183-6
Cheng, Lingyun; Chaidhawangul, Sunan; Wong-Staal, Flossie et al. (2002) Human immunodeficiency virus type 2 (HIV-2) vector-mediated in vivo gene transfer into adult rabbit retina. Curr Eye Res 24:196-201
Gruber, A; Wheat, J C; Kuhen, K L et al. (2001) Differential effects of HIV-1 protease inhibitors on dendritic cell immunophenotype and function. J Biol Chem 276:47840-3
Gruber, A; Looney, D J; Ibanez, M et al. (2001) Altered immunophenotype of dendritic cells generated from HIV infected subjects. Immunol Lett 78:209-11
Sauter, S L; Gasmi, M (2001) FIV vector systems. Somat Cell Mol Genet 26:99-129
Gruber, A; Kan-Mitchell, J; Kuhen, K L et al. (2000) Dendritic cells transduced by multiply deleted HIV-1 vectors exhibit normal phenotypes and functions and elicit an HIV-specific cytotoxic T-lymphocyte response in vitro. Blood 96:1327-33