This proposal from Dr. Mark Goldsmith from the Gladstone Institute of Virology is in response to a PA for proposals studying models of HIV disease and AIDS related malignancies. Since HIV infection occurs predominantly within lymphoid tissues in vivo, the investigator proposes to utilize the recently developed """"""""human lymphoid histocultures"""""""" or HLH with its' maintenance of tissue viability (up to 21 days), histologic architecture, cell-cell interactions and permissiveness to HIV infection as a model for preclinical investigation. In preliminary work, the investigator and his colleagues have been able to study the molecular and cellular aspects of lymphocyte depletion by HIV. In this application they propose to develop this system to """"""""produce a reliable, quantitative high-resolution and generalized model that is applicable to a variety of important aspects of preclinical investigation including anitviral drug development. In particular, the studies will focus on two essential virologic parameters - replication capacity (fitness) and pathogenic potential (virulence)"""""""". In SA 1, the influence of PI resistance mutations on the virulence and fitness properties of HIV will be assessed in HLH based on 1.1. ability of primary PIR and recombinant PIR to deplete CD4 cells in HLH 1.2. ability of PIR virus from patients with isolated virologic failure to deplete CD4 cells. 1.3 competitive assays of wild type and PIR virus in HLH but in the absence of drug to determine relative fitness (assessments will be made phenotypically using GHOST cells and genotypically using HTA). In SA2, HLH will be used to study whether evolution of HIV in vivo is associated with progressively greater virulence or viral fitness: 2.1 whether primary isolates obtained longitudinally exhibit differences in replication potential (viral load) or virulence (CD4 depletion) 2.2 whether naturally attenuated (Nef minus virus from the Sidney cohort) acquire enhanced replication or pathogenic potential through further genomic modifications. In SA 3, the impact of antiviral therapies on HIV fitness, virulence and genetic evolution will be studied in the HLH context: 3.1 define the rate and nature of genetic changes in HIV during replication in HLH with attention to co-receptor specificity in the absence of added selective pressure (assessment by assays with GHOST cells and HTA). 3.2 to determine antiviral therapies such as CXCR4 antagonist T22 or beta chemokines affect env evolution and Protease inhibitors in the case of evolution of protease. The PI has developed several """"""""advanced tools"""""""" for the HLH system including the ability to FACS based on CXCR4 and CCR5 expression to distinguish individual effects on these cells types and kinetic (time course) evaluations of cell depletion.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA086814-01
Application #
6077645
Study Section
Special Emphasis Panel (ZRG1-AARR-1 (01))
Program Officer
Read-Connole, Elizabeth Lee
Project Start
2000-07-01
Project End
2004-06-30
Budget Start
2000-07-01
Budget End
2001-06-30
Support Year
1
Fiscal Year
2000
Total Cost
$256,308
Indirect Cost
Name
J. David Gladstone Institutes
Department
Type
DUNS #
047120084
City
San Francisco
State
CA
Country
United States
Zip Code
94158
Roy, Ann-Marie; Schweighardt, Becky; Eckstein, Lauren A et al. (2005) Enhanced replication of R5 HIV-1 over X4 HIV-1 in CD4(+)CCR5(+)CXCR4(+) T cells. J Acquir Immune Defic Syndr 40:267-75
Jayakumar, Prerana; Berger, Irina; Autschbach, Frank et al. (2005) Tissue-resident macrophages are productively infected ex vivo by primary X4 isolates of human immunodeficiency virus type 1. J Virol 79:5220-6
Schaeffer, Evelyne; Soros, Vanessa B; Greene, Warner C (2004) Compensatory link between fusion and endocytosis of human immunodeficiency virus type 1 in human CD4 T lymphocytes. J Virol 78:1375-83
Hazarika, Parul; McCarty, Marya F; Prieto, Victor G et al. (2004) Up-regulation of Flotillin-2 is associated with melanoma progression and modulates expression of the thrombin receptor protease activated receptor 1. Cancer Res 64:7361-9
Stopak, Kim; de Noronha, Carlos; Yonemoto, Wes et al. (2003) HIV-1 Vif blocks the antiviral activity of APOBEC3G by impairing both its translation and intracellular stability. Mol Cell 12:591-601
Jekle, Andreas; Keppler, Oliver T; De Clercq, Erik et al. (2003) In vivo evolution of human immunodeficiency virus type 1 toward increased pathogenicity through CXCR4-mediated killing of uninfected CD4 T cells. J Virol 77:5846-54
Greene, Warner C; Peterlin, B Matija (2002) Charting HIV's remarkable voyage through the cell: Basic science as a passport to future therapy. Nat Med 8:673-80
Jekle, Andreas; Schramm, Birgit; Jayakumar, Prerana et al. (2002) Coreceptor phenotype of natural human immunodeficiency virus with nef deleted evolves in vivo, leading to increased virulence. J Virol 76:6966-73
Kreisberg, J F; Kwa, D; Schramm, B et al. (2001) Cytopathicity of human immunodeficiency virus type 1 primary isolates depends on coreceptor usage and not patient disease status. J Virol 75:8842-7
Penn, M L; Myers, M; Eckstein, D A et al. (2001) Primary and recombinant HIV type 1 strains resistant to protease inhibitors are pathogenic in mature human lymphoid tissues. AIDS Res Hum Retroviruses 17:517-23