In addition, neither BV nor CeHV-2 uses HVEM or African green monkey (AGM) (Chlorocebus sabaeus) HVEM as an entry receptor (Fig. Since the infection occurred in a living animal prior to analysis, we can conclude that tissue architecture and local defenses in the salivary gland or associated tissues have little or no discernible effect on dynamics of virus particle transport. All of the identified viral proteins are known virion components and were expected to be delivered into the axoplasm upon entry (26). In contrast, on complementing cells or in RK13 cells infected with the UL25 rescue mutant PrV-ΔUL25FR, all stages of capsid morphogenesis and virus assembly were easily observed and numerous mature virions lined the cell surface (Fig. Note that the EHV-1 primers were able to amplify cDNA from PRV 336-infected cells but not cDNA from cells infected with PRV 328 (negative control). 26;6(2):e1000785. Colored plaques were selected and purified on Vero cells.
The table on the right summarizes the cell surface expression of each construct when it is coexpressed with HSV-1 or SaHV-1 gL (first symbol) and the ability each construct to mediate fusion when it is coexpressed with entry glycoproteins from either HSV-1 or SaHV-1 (second symbol). 3B), and titers of 333d41R were lower than those of SB5 in the brains of 129 mice (P = 0.005) (Fig. Science 302: 1396-1398 Miller G, Lipman L (1973) Comparison of the yield of infectious virus from clones of human and simian lymphoblastoid lines transformed by Epstein-Barr virus. We thank Michael Coulthart, Morag Graham, and Sherrie Kelly at the National Microbiology Laboratory for helpful discussions on the analysis of the data for this article, Claire Sevenhuysen for a critical review of the manuscript, and Johnny Lee for technical assistance during his co-op term at the University of Victoria, Victoria, British Columbia, Canada. Overall, only 3 to 6% of capsids displayed as much as 25% overlap with gB, gD, or gE/gI, and these tended to be in regions of axons with dense concentrations of capsids and glycoproteins. These results indicate that during PRV infection, HDAC2 is posttranslationally modified in an additional US3 kinase-independent manner. Data were normalized to the fusion activity measured when a homotypic set of glycoproteins was coexpressed (either all HSV-1 or all SaHV-1 glycoproteins).
Neurons were explanted, infected with PrV-Ka, and analyzed 18 h after infection by immunoelectron microscopy as detailed in Materials and Methods. 3A). Suenaga, T., Kohyama, M., Hirayasu, K. RK13 cells were infected at an MOI of 1 with transcomplemented PrV-ΔUL25 and analyzed 14 h after infection. To determine whether PRV retrograde transport in axons depends on the Clip proteins, we knocked down Clip1 or Clip2 expression in chambered SCG neurons using small interfering RNA (siRNA) magnectofection. A digital counter will appear in the upper left corner. Detection of VZV peptide IE62915–929–reactive CD4 T cells in peripheral blood (PB)– and cord blood (CB) –derived mononuclear cells (MC) of HLA class II typed healthy adults and newborns, respectively.
Cells should not be piled on top of each other and axon bundling should be minimized. Horwitz and colleagues showed that EBV can infect human BMVECs, where it becomes latent . They should be controllable by vaccines, but despite notable effort, satisfactory vaccines remain unavailable. Rats were maintained in individual cages with food and water freely available throughout the experiment. (C) Still images from Movie S1, depicting a single viral exocytosis event. The suppressive effect of IFNalpha on PRV replication was sustained over a longer period of time since at 5dpi, still only 10±4% of infected neurons were late viral antigen positive (Fig. Virus replication starts up again, and infectious virions then travel back to the periphery, using the same sensory nerve “highway” used to reach the ganglia.
Instead, 561 nm DPSS lasers are typically used on commercial microscopes. Despite our ability to recover miR-H8, albeit infrequently, from subject 2, this miRNA was not detectable by qRT-PCR using RNA recovered from either subject 1 or subject 2. After fixing with PBS containing 4% paraformaldehyde and permeabilization with PBS containing 0.5% Triton X-100, the cells were stained with an anti-herpes simplex virus 1 (anti-HSV-1) antibody (B0114; Dako, Glostrup, Denmark) at 4°C overnight, followed by an Alexa Fluor 488-conjugated anti-rabbit IgG (Life Technologies) at room temperature for 30 min. The MEGA (version 5) program was used to construct multiple-sequence alignments on the basis of the amino acid sequences (34). Mouse MAbs that recognize PRV-specific proteins, gB (M2) and gC (M1), were described in a previous study (14). 1B, dark gray bars) or cells transfected with either GFP or mCherry alone contained, on average, 9 PML-NBs, whereas cells transfected with WT HSV-2 Us3 or HSV-2 Us3 fusion protein all contained, on average, 3 PML-NBs. Elevated mitochondrial motility in neurons infected with the gB-null strain was observed as late as 24 hpi, but was reduced at later times post infection.