The main goal of the Cell Culture Core facility is to provide a stable, reproducible and reliable source of virus stocks and relevant KSHV-infected and uninfected primary and immortalized cell cultures for all of the projects involved in this program. This centralized source would be cost-saving and provide for reproducible experimentation across the different laboratories. The Core will support all four projects with the following aims 1) produce, isolate and titer stocks of KSHV and recombinant KSHV strains 2) grow, maintain and provide KSHV-infected and noninfected tissue culture cells 3) process tonsil and oral KS biopsies for tissue sectioning or primary cultures 4) provide training for virus production and purification, cell culture growth and maintenance and for biosafety methods and procedures for producing and utilizing herpesviruses in research.

Public Health Relevance

The Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) is the cause of KS and two other AIDS-related malignancies. KS is the most common oral tumor associated with HIV/AIDS and is one of the most common pediatric and adult tumors in Sub-Saharan Africa. This project will investigate the immune control, activation, transmission and pathogenesis of KSHV in order to develop effective vaccines and therapeutics for this devastatino viral pathogen.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Program Projects (P01)
Project #
5P01DE021954-03
Application #
8463824
Study Section
Special Emphasis Panel (ZDE1-MH)
Project Start
Project End
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
3
Fiscal Year
2013
Total Cost
$168,033
Indirect Cost
$72,688
Name
Seattle Children's Hospital
Department
Type
DUNS #
048682157
City
Seattle
State
WA
Country
United States
Zip Code
98105
Garrigues, H Jacques; DeMaster, Laura K; Rubinchikova, Yelena E et al. (2014) KSHV attachment and entry are dependent on ?V?3 integrin localized to specific cell surface microdomains and do not correlate with the presence of heparan sulfate. Virology 464-465:118-33
Fontaine, Krystal A; Camarda, Roman; Lagunoff, Michael (2014) Vaccinia virus requires glutamine but not glucose for efficient replication. J Virol 88:4366-74
Garrigues, H Jacques; Rubinchikova, Yelena E; Rose, Timothy M (2014) KSHV cell attachment sites revealed by ultra sensitive tyramide signal amplification (TSA) localize to membrane microdomains that are up-regulated on mitotic cells. Virology 452-453:75-85
Gutierrez, Kimberley D; Morris, Valerie A; Wu, David et al. (2013) Ets-1 is required for the activation of VEGFR3 during latent Kaposi's sarcoma-associated herpesvirus infection of endothelial cells. J Virol 87:6758-68
Morris, Valerie A; Punjabi, Almira S; Wells, Robert C et al. (2012) The KSHV viral IL-6 homolog is sufficient to induce blood to lymphatic endothelial cell differentiation. Virology 428:112-20
DiMaio, Terri A; Gutierrez, Kimberley D; Lagunoff, Michael (2011) Latent KSHV infection of endothelial cells induces integrin beta3 to activate angiogenic phenotypes. PLoS Pathog 7:e1002424