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The innate immune response is an early line of defense that is essential for the detection of viruses. This means that you will not need to remember your user name and password in the future and you will be able to login with the account you choose to sync, with the click of a button.
This page doesn't support Internet Explorer 6, 7 and 8.Please upgrade your browser or activate Google Chrome Frame to improve your experience. However, whether this novel immune function of the dUTPase is specific for EBV or a common property of the Herpesviridae family is not known.
Cells contain a variety of sensors referred to as pattern recognition receptors (PRRs) that detect pathogen associated molecular patterns (PAMPs). Viruses from the three subfamilies contain a subset of genes that by criteria of genomic position and similarities in encoded amino acids sequences are common to all members. Wild-type pCMV-TLR1 vector was a gift from Dr Koichi Kuwano (Kurume University School of Medicine, Kurume, Japan).
Dharam Ablashi, HHV-6 Foundation), human herpesvirus-8 (HHV-8, ORF54; clone L54, which was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH from Drs.
Empty pCMV vector was used as a carrier to keep the total amount of transfected-DNA constant. These DCs express surface markers CD1a, HLA-DR, co-stimulatory molecules, Birbeck granules and surface markers characteristic of both plasmacytoid and myeloid DC (Ayehunie et al., 2003).


Following treatment, cell culture supernatants were collected and the levels of pro-inflammatory cytokines in treated and control samples were measured by ELISA as we have described (Ariza et al., 2013). A two-sample t-test was also used to compare cytokine levels among groups in the presence or absence of blocking antibody. Based upon the sensitivity (5 ng) of the EZBlue™ stain, the purified recombinant herpesviruses-encoded dUTPase proteins were estimated to be greater than 99% homogeneous (data not shown).
Salvatore Caradonna, Rowan University), varicella-zoster virus (VZV, ORF8) (Ross et al., 1997), as well as subcloning of the human dUTPase (kindly provided by Dr. Data were normalized for transfection efficiency by measuring Renilla luciferase activity and expressed as the mean relative stimulation ± SD.
Following treatment, cell culture supernatants were collected and the levels of cytokines in treated and control samples were measured by ELISA (MSD Multi-array and Multi-spot human cytokine kit) as we have described previously (Ariza et al., 2013). To our knowledge, this is the first report demonstrating that a non-structural protein encoded by herpesviruses HHV-6A, HHV-8, VZV and to a lesser extent HSV-2 is a pathogen-associated molecular pattern. IgG2a Isotype control monoclonal antibody was purchased from eBioscience (San Diego, California) and anti-TLR2 (clone TL2.1) monoclonal antibody was purchased from Imgenex (San Diego, California). The PCR product was purified using the QIAquick gel extraction kit (QIAGEN) and cloned into the protein expression vector pTrcHis Topo (Invitrogen).
Data were normalized for transfection efficiency by measuring Renilla luciferase activity and expressed as the mean fold induction ± SD relative to untreated control levels. Our results reveal a novel function of the virus-encoded dUTPases, which may be important to the pathophysiology of diseases caused by these viruses.


However, to the best of our knowledge there have not been any studies to determine whether non-structural proteins encoded by the human herpesviruses act as PAMPs and alter DC function. While the members of the human herpesvirus family have considerable diversity with respect to cellular tropism and pathogenesis, they all encode for putative dUTPases, suggesting that these proteins may be critical to the biology of these viruses. Twenty individual clones for each dUTPase construct type were isolated following transformation of E. Fractions were assayed for dUTPase activity as described previously (Glaser et al., 2006) and for protein using the Coomassie Brilliant Blue dye-binding assay (Bio-Rad Laboratories) and bovine serum albumin as the standard. Data were normalized for transfection efficiency by measuring Renilla luciferase activity and expressed as the mean fold induction ± SD relative to control levels. More importantly, this study demonstrates that the immunomodulatory functions of dUTPases are a common property of the Herpesviridae family and thus, the dUTPase could be a potential target for the development of novel therapeutic agents against infections caused by these herpesviruses. Furthermore, it is important to understand what virus-encoded macromolecules have the potential to modulate the innate and adaptive immune systems. In this report, we provide compelling evidence demonstrating that the immune modulatory properties of the monomeric dUTPases are not restricted to EBV, but rather it is a property of several members of the human herpesviruses. Proteins were visualized using EZBlue™ protein gel stain, as described by the manufacturer (Sigma Aldrich, St.



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