Thus, the packaging of viral DNA into C-capsids is accompanied by dramatic changes in the capsid association of some packaging proteins. We concluded that PDTC blocked HSV replication and gene expression through dysregulation of the cellular UPS. HSV-1 ICP27 and ICP34.5 have been shown to cause redistribution of p32 to the nucleus and nuclear rim, respectively (41, 42). Seventy-six patients were enrolled in the study, and their demographic features are summarized in Table 1⇓. Finally, we demonstrated that OT2 × RAG1−/− (OT2xRAG1−/−) mice, which are T cell transgenic to recognize an OVA peptide, still develop HSK; yet such mice failed to recognize any HSV antigens. The coverslips were further blocked in PBS containing 10% goat serum for 20 min and incubated for 20 min with anti-nucleoporin antibody MAb414 (Covance) diluted 1:2,000. Cells, viruses, and drugs.Primary human foreskin fibroblast (HFF), telomerase-immortalized human fibroblast (HFT) (19), and Vero cells were maintained in minimum essential alpha medium supplemented with 10% fetal calf serum (Gibco-Invitrogen).
Oligonucleotides.The phosphorothioate ODNs 5652 and fomivirsen, also known as ISIS 2922 (2922), were synthesized by Isis Pharmaceuticals and provided through the courtesy of Frank Bennett and Kevin Anderson. These findings show that natural products are still potential sources in the search for new antiherpetic agents. Louis, MO). The HSV-1 protein ICP47 inhibits CD8+ T-cell recognition of infected targets by impairing the transport of peptides into the endoplasmic reticulum for loading onto major histocompatibility complex (MHC) class I molecules and transport to the cell surface (1). As the insoluble nuclear lamina would present a significant barrier to capsid envelopment at the INM, it is logical that herpesviruses would devise or adopt means of modifying the nuclear lamina to promote nucleocapsid egress. However, the toxicity exhibited by an intact γ34.5 gene might reduce the potential application of the latter viruses as oncolytic agents. N.
Similarly, gD, gC, and gH were not found associated with these microdomains during infection. Our approach was to assess virus replication and markers of NFκB activation in mouse embryo fibroblasts (MEFs) with targeted deletions in key components of the signaling pathway (IKKα or IKKβ) or through overexpression of a DNIκBα and to assess HSV replication and the ability of HSV to suppress markers of apoptosis. The pre- and postfusion forms were also found on the surface of infected cells, but only the postfusion form was found in intracellular locations, suggesting that the protein initially folds in its postfusion conformation, is transported intracellularly in that form to avoid premature activity, and then refolds into the prefusion conformation when it reaches the plasma membrane (26). Moreover, surface expression of immature MHCII/invariant chain (Ii) complexes and the presence of these complexes in MVBs is elevated (5). The implications of these conclusions with respect to the restriction imposed on BHK cells by the expression of gD are discussed. The observation that herpes simplex virus (HSV) and human immunodeficiency virus bind heparan sulfate provided the rationale for the development of sulfated or sulfonated polymers as topical agents. To identify particular gB epitopes responsible for HSPG-independent binding, we used a panel of monoclonal antibodies (MAbs) to gB to block gB binding.
Together, these data favor a model in which HSV uses gB to rapidly mobilize lipid rafts that may serve as a platform for entry and cell signaling. Previous studies have led to the hypothesis that chromosomal alpha-globin genes are subject to chromatin-dependent repression mechanism that prevents expression in nonerythroid cells. This peptide-based strategy could lead to the identification of functionally important regions of gB or other membrane proteins and identify novel inhibitors of HSV-1 entry. The reaction of a nonprecipitating monoclonal antibody with electrophoretically separated, immobilized polypeptides contained in cytoplasmic and nuclear fractions, those chemically deglycosylated, or those specified by specific HSV-1 x HSV-2 intertypic recombinants identified the antigens reactive with the second monoclonal antibody as various forms of glycoprotein gC. 53) 54) Acanthamoeba infections are usually transmitted via insects. We report that UL13 is dispensable for viral replication in cell culture and is not involved in the processing of UL34 or of associated phosphoproteins. The compounds formed stable complexes with glycoprotein B and inhibit viral binding, entry, and cell-to-cell spread.
Our findings indicate that the widespread dispersion of MBCs to lymphoid tissues throughout the body is largely independent of the route of infection, but may be influenced by tissue-specific factors. Cell fusion caused by HSV-1 syn20, a mutant encoding syn gK, was suppressed when cells were coinfected with an Ad vector, AdgK, which expresses wild-type gK. Because the ectodomains of the wild-type and the mutant forms of gB crystallized at both low and neutral pH, we were able to determine the effect of pH on gB conformation at the atomic level. At low multiplicities of infection, viral gene expression in rabbit skin cells was delayed by many hours, although ultimately virus yield was comparable to that of the wild-type virus.