Dengue virus (DenV) is a devastating human pathogen that causes disease and viremia in millions of people worldwide every year. But very little is known about the initiation of immunity by DenV. In this proposal we will study the following aims:
Aim 1. What is the role of different populations of human DCs in the initiation of immunity by DenV? We hypothesize that there is a coordinated contribution of different cells in tissue and blood for the initiation/evasion of immunity and the establishment of infection by DenV. And while mosquito derived DenV may be able to inhibit important elements of innate immunity, such as type I IFN, subsequent human immune cell derived DenV may not be able to do so. This could explain the failure of DenV to transmit from human to human even though it causes viremia. We will analyze the pattern of activation and susceptibility to infection by DenV of different populations of APCs from human blood using both mosquito and primary human immune cell derived DenV. We will infect whole PBMCs to provide bystander effects and crosstalk between different cells which may resemble better the initial steps of initiation/evasion of immunity by DenV in vivo.
Aim 2. What is the contribution of individual DenV proteins to the maturational profile of DCs? We hypothesize that individual DenV proteins may be responsible for the induction/evasion of immunity in vivo. We will generate recombinant Newcastle disease virus (NDV) expressing individual DenV proteins and will infect DCs and other blood APCs. We will analyze the activation and cytokine/chemokines profile in those infected cells. Since NDV induces high levels of type I IFN production in those cells we may be able to detect still unknown IFN antagonists in DenV with this system. Additionally, these recombinant viruses will be used to generate monoclonal antibodies specific for non structural proteins of DenV.
Aim 3. What is the effect of DV infection or expression of individual DenV proteins on the T cell priming ability of different DCs towards Th1 immunity? DCs are the main APCs involved in priming of T cells in vivo after interaction with pathogens. Monocyte derived DCs infected with DenV have been shown to induce mixed Th0 responses in naive T cells. We hypothesize that the lack of type I IFN released by DenV infected APCs results in the generation of weak Th0 immunity in the host. We will evaluate the ability of different APCs from blood to secrete type I IFN and to prime Th1 or Th2 T cell responses after infection with DenV or with recombinant NDV expressing DV proteins. We will use qRT-PCR and multiplex ELISA for Th1/Th2 genes and cytokines/chemokines on T cells and APCs, with special emphasis on the levels of type I IFN released APCs. Dengue virus (DenV) can cause devastating disease in humans and there is no vaccine or treatment available to date. We are using a human system to study DenV initiation and evasion of immunity by analyzing the pattern of gene expression and cytokine and chemokine secretion profile of human dendritic cells (DCs)and other antigen presenting cells (APCs) and T cells after exposure to DenV. We will generate recombinant viruses expressing individual DenV proteins to analyze their role in the initiation/evasion of immunity by DenV. Understanding the modulation of cytokine responses by DenV, such as type I interferon (IFN), can help in the design of therapeutic targets to fight DenV.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI073450-05
Application #
8306324
Study Section
Special Emphasis Panel (ZRG1-IDM-B (02))
Program Officer
Cassetti, Cristina
Project Start
2008-08-15
Project End
2013-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
5
Fiscal Year
2012
Total Cost
$332,254
Indirect Cost
$136,234
Name
Icahn School of Medicine at Mount Sinai
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
078861598
City
New York
State
NY
Country
United States
Zip Code
10029
Manganaro, Lara; Pache, Lars; Herrmann, Tobias et al. (2014) Tumor suppressor cylindromatosis (CYLD) controls HIV transcription in an NF-?B-dependent manner. J Virol 88:7528-40
Opaluch, Amanda M; Schneider, Monika; Chiang, Chih-yuan et al. (2014) Positive regulation of TRAF6-dependent innate immune responses by protein phosphatase PP1-?. PLoS One 9:e89284
Rajsbaum, Ricardo; Versteeg, Gijs A; Schmid, Sonja et al. (2014) Unanchored K48-linked polyubiquitin synthesized by the E3-ubiquitin ligase TRIM6 stimulates the interferon-IKK? kinase-mediated antiviral response. Immunity 40:880-95
Brinzevich, Daria; Young, George R; Sebra, Robert et al. (2014) HIV-1 interacts with human endogenous retrovirus K (HML-2) envelopes derived from human primary lymphocytes. J Virol 88:6213-23
Morrison, Juliet; Laurent-Rolle, Maudry; Maestre, Ana M et al. (2013) Dengue virus co-opts UBR4 to degrade STAT2 and antagonize type I interferon signaling. PLoS Pathog 9:e1003265
Ramos, Irene; Krammer, Florian; Hai, Rong et al. (2013) H7N9 influenza viruses interact preferentially with *2,3-linked sialic acids and bind weakly to *2,6-linked sialic acids. J Gen Virol 94:2417-23
Suthar, Mehul S; Aguirre, Sebastian; Fernandez-Sesma, Ana (2013) Innate immune sensing of flaviviruses. PLoS Pathog 9:e1003541
Chiang, Chih-Yuan; Engel, Alex; Opaluch, Amanda M et al. (2012) Cofactors required for TLR7- and TLR9-dependent innate immune responses. Cell Host Microbe 11:306-18
Rodriguez-Madoz, Juan R; Belicha-Villanueva, Alan; Bernal-Rubio, Dabeiba et al. (2010) Inhibition of the type I interferon response in human dendritic cells by dengue virus infection requires a catalytically active NS2B3 complex. J Virol 84:9760-74
Rodriguez-Madoz, Juan R; Bernal-Rubio, Dabeiba; Kaminski, Dorota et al. (2010) Dengue virus inhibits the production of type I interferon in primary human dendritic cells. J Virol 84:4845-50

Showing the most recent 10 out of 11 publications