The principal goal of this project is to understand the impact of HIV envelope-mediated signaling on viral replication and immune dysfunction. This information may facilitate the discovery of new strategies for the treatment of HIV infection. HIV envelope proteins, which are displayed on the surface of viral particles, bind to two receptors on T-lymphocytes in order to facilitate infection of those cells. Initially envelope proteins on the outer surface of virions dock with the CD4 receptor, and subsequently engage a co-receptor, either CCR5 or CXCR4. Envelope engagement of each of these receptors results in signal transduction; however the consequences of this signaling is only partially understood. In previous studies we demonstrated that HIV envelope-mediated signaling induces several biological responses in primary T-cells and macrophages, including the induction of proinflammatory cytokines and increased rate of apoptosis. More recently it has been shown that envelope also binds to DC-SIGN, a C-type lectin receptor. We have determined that gp120 binding to DC-SIGN on dendritic cells also mediates biological responses. We evaluated the response of monocyte derived dendritic cells (MDDCs) to a panel of recombinant oligomeric envelope proteins with different glycosylation content. Modulation of gene expression was evaluated using oligonucleotide microarrays, and protein phosphorylation events were evaluated with high throughput proteomic analysis. MDDC responses were strongly influenced by the carbohydrate content of gp120. We identified genes upregulated only by a hyperglycosylated gp120 and genes upregulated only by a hypoglycosylated gp120. We are also evaluating gp120 interactions with other C-type lectin receptors that are expressed on other cell types. ? Understanding how various cell types respond to gp120 may provide further insight into the basis of HIV induced immune dysfunction and pathogenesis. Of particular interest is the interaction of gp120 with plasmacytoid dendritic cells (pDCs). pDCs, which play a central role in innate and adaptive immune responses against viral infections, secrete type I IFNs and proinflammatory cytokines upon stimulation via either TLR7 or TLR9. Throughout the course of HIV infection, the production of type-I IFNs is profoundly impaired, and total pDC cell counts in peripheral blood correlates inversely with viral load and positively with CD4+ T-cell count. The origin of these defects is unclear. pDCs express CD4, CCR5 and CXCR4, the primary receptor and co-receptors, respectively, for the HIV envelope, yet little is known about the effects of the viral envelope on these cells. We determined that exposure of pDCs to gp120 results in the suppression of activation of these cells. This suppression was specific for TLR9-mediated responses, since TLR7-mediated responses were unaffected by gp120. gp120 also suppressed TLR9-mediated induction of proinflammatory cytokines and the expression of CD83, a marker of DC activation. Finally, gp120 suppressed pDC-induced cytolytic activity of NK cells. Taken together, these data demonstrate that the direct interaction of HIV-1 gp120 with pDCs interferes with TLR9 activation resulting in a decreased ability of pDCs to secrete antiviral and inflammatory factors that typically play a central role in initiating host immune responses against invading pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000887-07
Application #
7592258
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2007
Total Cost
$853,757
Indirect Cost
City
State
Country
United States
Zip Code
Kelley, James M; Daley, George Q (2013) Hematopoietic defects and iPSC disease modeling: lessons learned. Immunol Lett 155:18-20
Garron, Marie-Line; Arthos, James; Guichou, Jean-Francois et al. (2008) Structural basis for the interaction between focal adhesion kinase and CD4. J Mol Biol 375:1320-8
Arthos, James; Cicala, Claudia; Martinelli, Elena et al. (2008) HIV-1 envelope protein binds to and signals through integrin alpha4beta7, the gut mucosal homing receptor for peripheral T cells. Nat Immunol 9:301-9
Martinelli, Elena; Cicala, Claudia; Van Ryk, Donald et al. (2007) HIV-1 gp120 inhibits TLR9-mediated activation and IFN-{alpha} secretion in plasmacytoid dendritic cells. Proc Natl Acad Sci U S A 104:3396-401
Cocklin, Simon; Gopi, Hosahudya; Querido, Bianca et al. (2007) Broad-spectrum anti-human immunodeficiency virus (HIV) potential of a peptide HIV type 1 entry inhibitor. J Virol 81:3645-8
Cicala, Claudia; Arthos, James; Censoplano, Nina et al. (2006) HIV-1 gp120 induces NFAT nuclear translocation in resting CD4+ T-cells. Virology 345:105-14
Cicala, Claudia; Arthos, James; Martinelli, Elena et al. (2006) R5 and X4 HIV envelopes induce distinct gene expression profiles in primary peripheral blood mononuclear cells. Proc Natl Acad Sci U S A 103:3746-51
Snyder, Greg A; Ford, Jennifer; Torabi-Parizi, Parizad et al. (2005) Characterization of DC-SIGN/R interaction with human immunodeficiency virus type 1 gp120 and ICAM molecules favors the receptor's role as an antigen-capturing rather than an adhesion receptor. J Virol 79:4589-98
Gupta, Neil; Arthos, James; Khazanie, Prateeti et al. (2005) Targeted lysis of HIV-infected cells by natural killer cells armed and triggered by a recombinant immunoglobulin fusion protein: implications for immunotherapy. Virology 332:491-7
Wang, Shixia; Arthos, James; Lawrence, John M et al. (2005) Enhanced immunogenicity of gp120 protein when combined with recombinant DNA priming to generate antibodies that neutralize the JR-FL primary isolate of human immunodeficiency virus type 1. J Virol 79:7933-7

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