Herpes simplex virus 1 recombinant virions exhibiting the amino terminal fragment of urokinase-type plasminogen activator

Expression of HSV-1 proteins in recombinant baculovirus-infected insect cells.The UL28 gene was subcloned as aHindIII-BamHI fragment from the pECH82 plasmid (63) (kindly provided by Nels Pederson) into theHindIII and BglII sites of baculovirus transfer vector pVL1393 (Pharmingen). Anti-HSV VP5 antibody was from Abcam (Cambridge, MA). Plasmids were constructed for this study as follows. The presence of C pneumoniae or CMV independently was associated with a greater risk of thrombosis on the plaques but not plaque ulceration. Kapp (Emory University, Atlanta, Ga.) (22). For monitoring the progress of infection and imaging, the dishes were transferred to a heated stage of the inverted LSM410 confocal microscope and monitored at different times thereafter. Cell viability was determined by the CellTiter-Glo assay for ATP (G-7570; Promega, Madison, WI), performed according to the manufacturer’s instructions.

Cells were then washed once with DMEM and infected with wild-type KOS at a multiplicity of infection between 0.01 and 0.1. After solvent was removed, the crude extracts were dissolved in dimethyl sulfoxide (DMSO) to a concentration of 100 mg/ml and stored at 4°C until use. Recombinant influenza virus bearing hemagglutinin and neuraminidase from the strain A/Brisbane/59/2007 (the CDC-recommended vaccine strain for the 2008/2009 and 2009/2010 seasons) and the rest of the genomic segments from A/Puerto Rico/08/1934 (BB.PR8) was kindly provided by Adolfo Garcia-Sastre. We investigated whether this kinetic change of gB expression would influence HSV-1 virulence, the generation and homing of gB-CD8 cells to the TG, and the ability of gB-CD8 cells to inhibit HSV-1 reactivation from latency. Furthermore, these various UL34-UL31 homologous pairs share a number of characteristics: (i) they are codependent for proper targeting to the nuclear rim; (ii) they facilitate nuclear egress of their respective nucleocapsids; and (iii) they share the capacity to interact with, and to induce structural alterations to, the nuclear lamina. Weller, University of Connecticut. Reagents.N-9 and chondroitin sulfate C were purchased from Sigma, St.

HSV-1 (KOS) and an HSV-1 strain (KOS/tk12) that carries the lacZ gene under the control of the ICP4 promoter (80) were both purified on sucrose gradients as described elsewhere (26). After 1 h, monolayers were covered with DMEM-H containing 2% calf serum and 0.3% methylcellulose. If low pH can indeed trigger an activating conformational change in gB, several possibilities can be envisaged for how inappropriate membrane fusion might be avoided during transport of the protein in infected cells. For cell surface expression of MHCII, backfusion of the MVB internal vesicles with the limiting membrane and subsequent amalgation of MVBs with the cell membrane was suggested (8). Among the important steps in infection, attachment and entry have attracted attention as targets for therapeutic treatments and vaccines. Accumulated evidence indicates that plasma membrane microdomains, or lipid rafts, that are highly enriched in cholesterol and sphingolipids play a crucial role in the lateral organization of the plasma membrane (9, 27, 71). Mailing address: Mount Sinai School of Medicine, Pediatric Infectious Diseases, 1 Gustave L.

(ii) Expression of wild-type virus genes in these cells precluded the formation of DNA-protein structures that would be affected by either the HDACs or their inhibitors. Importantly, both US3 and gB are required for efficient inhibition of CD1d antigen presentation and NKT cell activation. Binding of gB to DR competes with binding to Ii. INTRODUCTION Herpes simplex virus 1 (HSV-1) virions have three morphologically distinct structures: the nucleocapsid, an icosahedral capsid containing the 152-kbp double-stranded DNA viral genome encoding at least 84 viral proteins; the tegument, a proteinaceous layer surrounding the nucleocapsid; and the envelope, a lipoprotein membrane with a host cell-derived lipid bilayer enclosing the nucleocapsid and tegument (1). Finally, vaccination prevented death in 83% of the animals challenged with a HSV-2 dose that killed 78 and 100% naive and mock-vaccinated controls, respectively. We conclude the following: (i) in cells infected with the compensatory mutants, US11 made early in infection binds to PKR and precludes the phosphorylation of eIF-2alpha, whereas US11 driven by its natural promoter and expressed late in infection is ineffective; and (ii) activation of PKR by double-stranded RNA is a common impediment countered by most viruses by different mechanisms. Similarly, ISIS 5652 was able to inhibit entry of pre-attached virions into cells at 37 degrees C, but the mutant did not exhibit resistance in this assay.

Electron microscopic studies revealed that HBsAg harvested from the extracellular medium and banded in CsCl density gradients contained spherical particles 15-22 nm in diameter, characteristic of empty HBV envelopes. This process apparently requires prior completion of the DNA packaging process. A biopsy specimen of the lesions revealed herpes simplex virus. It suppresses CD1d expression primarily by inhibiting its recycling to the cell surface after endocytosis. Multiple mar mutations were also introduced into the gB structural gene by recombination and sequential antibody selection to produce a set of mar mutants with double, triple, and quadruple epitope alterations. We also tested 14 retrocyclin analogues, including the retro, enantio, and retroenantio forms of retrocyclin 1.