Alphaviruses are arthropod-vectored viruses that can cause human diseases ranging from mild febrile illness to severe arthralgia or fatal encephalitis. Due to several factors including the ease of their isolation, long-term stability and potential for high morbidity/mortality when administered by aerosol, several have been designated Category B Select Agents with potential for biowarfare/bioterrorism use. Unfortunately, no licensed vaccines or antiviral medications are available to combat these infections. Interruption of the initial interactions of viruses with cells is one of the most effective means of preventing infection and disease and an understanding of virus-infection receptor interactions is vital to design of infection-blocking drugs. However, little is known about the receptor interactions of alphaviruses that have not been adapted extensively to growth in cell culture. Using a novel vector system, we propose to examine simultaneously, the receptor interactions of multiple non-cell adapted alphaviruses representative of both New World and Old World strains. In the first Aim, we will investigate the utilization of previously characterized cell surface receptor molecules heparan sulfate and the high affinity laminin receptor using well-characterized cell lines. In the second Aim we will utilize our novel vector system and a recently-identified alphavirus receptor-negative cell line in a cDNA library screen to identify new molecules that can mediate alphavirus infection of cells. The results of these experiments will provide a comprehensive understanding of the receptor interactions of New World and Old World alphaviruses and provide a framework for design of infection-blocking antiviral drugs.

Public Health Relevance

Our plan is to use a novel chimeric alphavirus replicon system in which a propagation-defective Venezuelan equine encephalitis virus genome is packaged in the structural proteins of different Old World and New World alphaviruses to compare the receptor utilization characteristics of low passage strains of these viruses. These comparisons will include direct evaluation of the capacity of previously-identified receptors (heparan sulfate, the high affinity laminin receptor) to mediate infection as well characterization of new receptors through use of a retrovirus expression library and alphavirus receptor-negative cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
7R21AI072350-03
Application #
7990854
Study Section
Special Emphasis Panel (ZRG1-IDM-P (91))
Program Officer
Repik, Patricia M
Project Start
2008-07-01
Project End
2011-06-30
Budget Start
2009-11-01
Budget End
2011-06-30
Support Year
3
Fiscal Year
2009
Total Cost
$183,125
Indirect Cost
Name
University of Pittsburgh
Department
Type
Organized Research Units
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Gardner, Christina L; Burke, Crystal W; Higgs, Stephen T et al. (2012) Interferon-alpha/beta deficiency greatly exacerbates arthritogenic disease in mice infected with wild-type chikungunya virus but not with the cell culture-adapted live-attenuated 181/25 vaccine candidate. Virology 425:103-12
Gardner, Christina L; Ebel, Gregory D; Ryman, Kate D et al. (2011) Heparan sulfate binding by natural eastern equine encephalitis viruses promotes neurovirulence. Proc Natl Acad Sci U S A 108:16026-31
Yin, Jun; Gardner, Christina L; Burke, Crystal W et al. (2009) Similarities and differences in antagonism of neuron alpha/beta interferon responses by Venezuelan equine encephalitis and Sindbis alphaviruses. J Virol 83:10036-47
Burke, Crystal W; Gardner, Christina L; Steffan, Joshua J et al. (2009) Characteristics of alpha/beta interferon induction after infection of murine fibroblasts with wild-type and mutant alphaviruses. Virology 395:121-32
Gardner, Christina L; Yin, Jun; Burke, Crystal W et al. (2009) Type I interferon induction is correlated with attenuation of a South American eastern equine encephalitis virus strain in mice. Virology 390:338-47
Meier, Kathryn C; Gardner, Christina L; Khoretonenko, Mikhail V et al. (2009) A mouse model for studying viscerotropic disease caused by yellow fever virus infection. PLoS Pathog 5:e1000614