Binding of herpes simplex virus glycoprotein D to nectin-1 exploits host cell adhesion. – PubMed

Recombinant ectodomain gD1 was scaled up and purified from the culture medium using nickel nitrilotriacetic acid affinity chromatography, and a maximum production level of 56.8 mg/L of recombinant gD1 was obtained in a shake-flask culture of S2 cells after induction with 5 µM CdCl2 for 4 days. However, binding of MAbs to the AAA protein or to single mutants altered at site 1 was reduced compared with TM-gD. This was due to the key role played by Y38 in interacting with nectin-1. Plaque production of F-gD beta particles lacking gD was enhanced by polyethylene glycol treatment, suggesting that gD is essential for penetration of HSV into cells. Together these data are consistent with previous studies showing that gD disrupts the normal nectin-1 homophilic interactions. Two mutant proteins, Cys-2 (Cys-106 to Ser) and Cys-4 (Cys-127 to Ser), were able to complement F-gD beta at 31.5 degrees C but not at 37 degrees C. The antigenic conformation of QAA was similar to that of gD produced in the presence of tunicamycin (TM-gD).

The gEctg, gEctg+gMctg, gDΔct, and gEctg+gDΔct viruses produced viral plaques on African monkey kidney cells (Vero), as well as other cells, that were on average approximately 30 to 50% smaller than those produced by the wild-type virus HSV-1(F). Ions and some metal complexes are excluded, as well as cases where no interaction profile could be generated. Thus, the affinity data follow the same hierarchy as the blocking data. Our results point to a binding hot spot centered around HveA-Y23, a residue that protrudes into a crevice on the surface of gD. During HSV infection, nectin-1 localization at adherens junction was dramatically altered in a manner dependent on gD expression. MAbs that recognize antigenic sites Ib and VII of gD were the only MAbs which blocked the HSV-HVEM interaction. Johnson, R.

For example, if the current year is 2008 and a journal has a 5 year moving wall, articles from the year 2002 are available. (B) An enlarged view of the gD-HveA interface shown in the same orientation as in panel A. For example, if the current year is 2008 and a journal has a 5 year moving wall, articles from the year 2002 are available. It is now clear that, on some cells, such as CHO and HeLa, HSV entry occurs by endocytosis (36, 37). We show here that gD from PrV and BHV-1 binds directly to the human receptors that mediate PrV and BHV-1 entry. For example, if the current year is 2008 and a journal has a 5 year moving wall, articles from the year 2002 are available. The mutations that resulted in failure to block apoptosis were the same for gD−/− and gD−/+ viruses and were located in three sites, one within the immunoglobulin-type core region (residues 125, 126, and 151), one in the upstream connector region (residues 34 and 43), and one in the C-terminal portion of the ectodomain (residue 277).

For example, if the current year is 2008 and a journal has a 5 year moving wall, articles from the year 2002 are available. HSV cellular receptor has been immobilized on the surface of the biosensor cuvette, bearing a carboxymethyl dextran layer. Both PFD subdomains bound gD260 t, highlighting multiple contact sites between the N and C termini of gD. These are the first mutations of the gene encoding gD of HSV identified in vivo in human encephalitis samples. of HSV2 via the footpad route were significantly protected against infection at all doses tested when compared with unimmunized AlPO4 and uninoculated control animals. Immunization with the recombinant virus also protected the majority of the mice against the development of a latent HSV-1 infection of the trigeminal ganglia. The Bac-to-Bac expression system was used to express HSV-2 gD in insect cells.

Sera from animals immunized with the synthetic peptide reacted with native gD and neutralized both HSV-1 and HSV-2. Recombinant baculoviruses have been widely used as safe vectors to express heterologous genes in the culture of insect cells, but the manipulation involved in creating, titrating, and amplifying viral stocks make it time consuming and laborious. We examined the efficiency and receptor specificity of this activity and used sequential incubation protocols to determine the order and stability of the initial interactions required for entry. Animals given gD-2t in SAF-m had higher anti-gD-2t antibodies, fewer and less severe vaginal lesions, and decreased ganglionic latency compared to animals given gD-2t in saline. was investigated for both its inactivating effects on some viruses and its antiviral effects against the viruses in vitro. In the present study, we investigated whether animals which had been functionally depleted of T-cell subsets or asialo GM1+ cells would continue to be responsive to MAb therapy. A live attenuated HSV-1 (deleted in the gE gene), and a HSV-1 (strain HSZP)-infected cell extract served as positive controls, and three non-structural recombinant HSV-1 fusion proteins (ICP27, UL9/OBP and thymidine kinase–TK) were used as presumed non-protective (negative) controls.

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Fatal disseminated cercopithecine herpesvirus 1 (herpes B infection in cynomolgus monkeys (Macaca fascicularis). – PubMed

Serum samples were collected from resident animals within this room at the time of presentation of case 2. Further, since the rhesus LCV was able to infect human B cells, the species restriction was after virus penetration (Moghaddam et al., 1998). B virus antibody-positive (n = 75) or -negative (n = 40) rhesus and cynomolgus serum samples were randomly selected for assessments of macaque serum reactivities with recombinant proteins. Despite the sequence variation in the UL39 ORF, alignment of only five sequence pairs was required before no further reduction in SSDs occurred. In order to determine if a B virus isolate was resistant to ACV or GCV, we created two thymidine kinase-defective B viruses as controls (Fig. This test has now been used over a three-year time period for routine screening of all blood, biopsy and necropsy samples submitted to the NZP Elephant Herpesvirus laboratory. In the ovarian histopathology study, monkey no.

Thus, only a single copy of the putative LAT region plus the RL2 (ICP0) and RL1 (γ34.1) ORFs are present in the HVS1 genome. Again, paralysis ED50 values for CDV were similar among the 3 viruses and were approximately twice that for protection against death. Radiolabeled extracts from COS7 cells transfected with the pBgD (D) or pcDNA vector DNA (V) were immunoprecipitated with various sera (dilution, 1:50). The advanced age of the 2 macaques affected may have predisposed them to infection with SVV; however, other geriatric rhesus housed in the same room remained uninfected. Furthermore, although it is rare, nonexperimental horizontal transmission of SIV has occurred between macaques and therefore continued SIV monitoring of SPF colonies associated with AIDS research centers is warranted. Statistical associations between macaque viral seropositivity, sex, age class, and group number were determined by χ2 test. Cage furnishings should be routinely inspected for broken and hazardous items that need to be repaired or replaced.

1C). 2005. These … In D. The UL29 nucleotide sequence reported here is >99% identical to a FPTHV UL29 sequence in GenBank (27). CACACA sequences are also found in intergenic DNA. The resulting peptides were analyzed by LC/MS/MS, and the data were correlated against the MHV-68 and NCBI mouse databases using SEQUEST.

To confirm the size of the inverted repeats flanking the unique regions, both virion and nucleocapsid DNA were digested by BamHI and EcoRI to ensure all possible isomeric forms of the viral genome were included (). A complete list of oligonucleotides and primers for all EMSAs can be found in the supplemental material. Some viral genes showing no recognizable similarity to host genes might occur due to deletions and insertions of different parts of coding districts accompanied by numerous single-nucleotide substitutions fixed by selection or genetic drift. There is no clear-cut distinction between latency and the slow-and-low approach. BatBHV-1 is most similar to TuHV-1(61%). The most extensive regions of predicted structure in introns occur within a repetitive region of the EBV genome (the W repeats). Gaps in regions not covered by spanning clones were closed using conventional PCR.

Serum and plasma samples.Plasma samples were obtained during the biannual health screening of macaques at the WaNPRC. ). For animals in an agricultural setting, AAALAC International takes the position that, in accredited programs, the housing and care for farm animals should meet the standards that prevail on a high-quality, well-managed farm. For instance, no clear cut switches of polarity were observed in the GC skew plot. Academically, she served as an intern at Zoo Atlanta, a volunteer research assistant at White River National Refuge, a volunteer research assistant at Yerkes Primate Center, and was the National Science Foundation Undergraduate Research Fellow. Macacine herpesvirus 1 (monkey B virus; BV) is an α-herpesvirus that is indigenous in macaque monkeys (Macaca spp.). Reviews of injuries and biohazard exposures among workers exposed to NHPs suggest that mucocutaneous contact with NHP body fluids is common; in one survey, 16 of 17 contacts with primate body fluids involved ocular exposure.

The genomes are linear, double-stranded DNA ranging from 125–241 kb and have a guanine + cytosine content of 32–75 % (McGeoch et al., 2006). Both animals had peritoneal effusions, massive necrosis of pharyngeal, esophageal, and gastric mucosa, and multifocal hepatic and pancreatic necrosis. This investigation documented the hazard of ocular splashes and indicated that dendritic corneal lesions, such as herpetic skin vesicles, are not always present in B virus infection. Cercopithecine herpesvirus 1 (B virus), an alphaherpesvirus endemic in Asian macaques, is closely related to herpes simplex virus (HSV). Cercopithecine herpesvirus 1 (monkey B virus; BV) produces extremely severe and usually fatal infections when transmitted from macaque monkeys to humans. This case report describes a rhesus macaque (Macaca mulatta; male; age, 5 y; weight, 6.7 kg) with anorexia, dehydration, lethargy, ataxia, and generalized skin rashes that occurred 30 d after total-body irradiation at 6.5 Gy ((60)Co γ-rays).

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