The genome of the human immunodeficiency virus type 1 (HIV-1) is highly heterogeneous. Some of this genomic variability most likely translates into the biologic and serologic differences observed among various strains of HIV-1. HIV-1 isolates vary in their ability to infect different cell types ( e.g. established T lymphoid cell lines, primary macrophages, human fibroblast cells), to replicate to high titers, and to cause damage to the infected cell. Difference in the isolates' susceptibility to serum neutralization have also been observed. Some of these in vitro differential properties have been correlated with pathogenicity of the virus in vivo. Defining the functional gene(s) of HIV-1 that control these biologic and serologic properties could help in the understanding of HIV-1 pathogenesis and in designing strategies for anti-viral therapies. This proposal is aimed at identifying the genes and domains within genes that determine the host range, cytopathogenicity and sensitivity to serum neutralization of HIV-1. Results obtained from our preliminary studies with recombinant viruses generated between different biologically active HIV-1 clones indicate that viral entry, cytopathology and sensitivity to neutralization are determined by the envelope gene, whereas kinetics of replication appear to be controlled by the LTR region. We propose to generate additional recombinant viruses between parental clones already used in our preliminary studies in order to define functional domains within the HIV-1 envelope gene and LTR that affect these biologic and serologic properties (Specific aims #1). We will also molecularly clone and sequence other HIV-1 isolates of interest (Specific aim #2) for the construction of new recombinant viruses (Specific #3). These will be useful in further verification of those genes and domains defined in Specific aim #1, and in the potential identification of additional regions of functional significance. The effects of specific changes within the functional domains will be investigated by site-directed mutagenesis (Specific #4). Finally, in vitro transient gene expression assays (Specific aim #5) will be used to explore further the role of the viral LTR and tat gene in determining the host range of HIV-1.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI029857-02
Application #
3144801
Study Section
Special Emphasis Panel (ARR (V1))
Project Start
1990-04-01
Project End
1993-03-31
Budget Start
1991-04-01
Budget End
1992-03-31
Support Year
2
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Stamatatos, L; Zolla-Pazner, S; Gorny, M K et al. (1997) Binding of antibodies to virion-associated gp120 molecules of primary-like human immunodeficiency virus type 1 (HIV-1) isolates: effect on HIV-1 infection of macrophages and peripheral blood mononuclear cells. Virology 229:360-9
Koito, A; Stamatatos, L; Cheng-Mayer, C (1995) Small amino acid sequence changes within the V2 domain can affect the function of a T-cell line-tropic human immunodeficiency virus type 1 envelope gp120. Virology 206:878-84
Harrowe, G; Cheng-Mayer, C (1995) Amino acid substitutions in the V3 loop are responsible for adaptation to growth in transformed T-cell lines of a primary human immunodeficiency virus type 1. Virology 210:490-4
Stamatatos, L; Cheng-Mayer, C (1995) Structural modulations of the envelope gp120 glycoprotein of human immunodeficiency virus type 1 upon oligomerization and differential V3 loop epitope exposure of isolates displaying distinct tropism upon virion-soluble receptor binding. J Virol 69:6191-8
Koito, A; Harrowe, G; Levy, J A et al. (1994) Functional role of the V1/V2 region of human immunodeficiency virus type 1 envelope glycoprotein gp120 in infection of primary macrophages and soluble CD4 neutralization. J Virol 68:2253-9
Stamatatos, L; Werner, A; Cheng-Mayer, C (1994) Differential regulation of cellular tropism and sensitivity to soluble CD4 neutralization by the envelope gp120 of human immunodeficiency virus type 1. J Virol 68:4973-9
LeGuern, M; Shioda, T; Levy, J A et al. (1993) Single amino acid change in Tat determines the different rates of replication of two sequential HIV-1 isolates. Virology 195:441-7
Stamatatos, L; Cheng-Mayer, C (1993) Evidence that the structural conformation of envelope gp120 affects human immunodeficiency virus type 1 infectivity, host range, and syncytium-forming ability. J Virol 67:5635-9
Cheng-Mayer, C; Shioda, T; Levy, J A (1991) Host range, replicative, and cytopathic properties of human immunodeficiency virus type 1 are determined by very few amino acid changes in tat and gp120. J Virol 65:6931-41
Liu, Z Q; Wood, C; Levy, J A et al. (1990) The viral envelope gene is involved in macrophage tropism of a human immunodeficiency virus type 1 strain isolated from brain tissue. J Virol 64:6148-53