HSV-1 Infection Inhibits Procollagen and Protein Secretion from Normal and Ataxia Telangiectasia Cultured Skin Fibroblasts (original) (raw)
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Journal of Clinical Investigation, 1995
The mechanisms that regulate the expression of genes encoding extracellular matrix proteins in fibroblasts and other mesenchymal cells have remained elusive. Studies from several laboratories have indicated that Tax, a trans-regulatory protein from the human T cell leukemia virus type I not only augments viral gene expression but also triggers the expression of various cellular genes. Here, we examined the hypothesis that the expression of collagen genes may also be modulated by Tax. NIH-3T3 cells were simultaneously transfected with a Tax expressor plasmid and a chimeric construct containing regulatory sequences (-804 to +42 bp) of the al(I) procollagen gene (COLlAl) promoter. The results indicated that the promoter activity of the -804 to +42 bp COLlAl fragment increased up to 12-fold in cells expressing Tax. Deletion analysis revealed that the region of COLlAl encompassing nucleotides -174 to -84 contained the Tax-responsive elements. A gene segment encompassing nucleotides -187 to -67, which contained this region, proved sufficient to confer Tax inducibility (2.5-fold) to a herpes simplex virus thymidine kinase promoter. Stably transfected NIH-3T3 cell clones that constitutively produce Tax displayed elevated levels of al (I) procollagen and fibronectin transcripts and increased production and accelerated processing of type I procollagen. These findings suggest that retroviral proteins may be involved in the pathogenesis of idiopathic diseases accompanied by collagen overproduction. (J. Clin. Invest. 1995. 96:2413-2420
Laminin Reduces HSV-1 Spread from Cell to Cell in Human Keratinocyte Cultures
Biochemical and Biophysical Research Communications, 1997
Herpes simplex virus-1 (HSV-1) infects epidermal cells in 1, 3, 5, 6). where it replicates and spreads from cell to cell. While While viral factors responsible for cell-to-cell spread some of the viral factors responsible for cell-to-cell have been identified, little is known about the cellular spread are known, the host cell molecules and structures factors which are utilized by HSV-1 in order to spread. which are utilized by HSV-1 during spread are not well A better understanding of the mechanism of cell-to-cell studied. Here we report that a laminin substrate respread of HSV-1 will require examination of these host duced the ability of HSV-1 to spread from cell to cell factors. in cultures of a human keratinocyte cell line (HaCaT). Cell-to-cell spread of viruses including HSV-1 can be Laminin did not reduce spread of the virus by decreasdetermined and measured by the morphology and size ing the viral replication rate. However, laminin did stimof plaques of infected cells formed in epidermal cell ulate the formation of tight junctions between HaCaT cultures (3, 7, 8). The plaque assay requires a monocells, suggesting that tight junctions can affect cell-tolayer of cells which has been inoculated at a low multicell spread of HSV-1. Since laminin is an abundant complicity of infection (M.O.I.). After initial infection of the ponent of the basement membrane in vivo, culturing cells, a neutralizing antibody can be added to the culcells on laminin may provide an assay which more accurately reflects the rate and mechanism of HSV-1 cell-to-ture medium to prevent cell-free virus from continuing cell spread in vivo.
Protease, Growth Factor, and Heparanase-Mediated Syndecan-1 Shedding Leads to Enhanced HSV-1 Egress
Viruses
Heparan sulfate (HS) and heparan sulfate proteoglycans (HSPGs) are considered important for the entry of many different viruses. Previously, we demonstrated that heparanase (HPSE), the host enzyme responsible for cleaving HS chains, is upregulated by herpes simplex virus-1 (HSV-1) infection. Higher levels of HPSE accelerate HS removal from the cell surface, facilitating viral release from infected cells. Here, we study the effects of overexpressing HPSE on viral entry, cell-to-cell fusion, plaque formation, and viral egress. We provide new information that higher levels of HPSE reduce syncytial plaque formation while promoting egress and extracellular release of the virions. We also found that transiently enhanced expression of HPSE did not affect HSV-1 entry into host cells or HSV-1-induced cell-to-cell fusion, suggesting that HPSE activation is tightly regulated and facilitates extracellular release of the maturing virions. We demonstrate that an HSPG-shedding agonist, PMA; a prot...
Journal of Clinical Investigation, 1993
CD8' cytotoxic T lymphocytes (CTL) clones with specificity for herpes simplex virus (HSV) were derived from two donors with genital HSV-2 infection. These CIL clones specifically lysed HSV-infected autologous B lymphoblastoid cells, but not HSV-infected fibroblasts. Exogenous peptide loading sensitized both cell types to lysis by an HSV-specific ClL clone of known specificity. HSV infection rendered fibroblasts refractory to peptide sensitization. HSV infection also rendered fibroblasts and keratinocytes insensitive to lysis by allospecific CD8 + CI'L clones. Lysis of B lymphoblastoid cells in this system was only slightly reduced by HSV infection. Reduction of fibroblast allospecific lysis was dose and time dependent and was blocked by acyclovir, indicating the involvement of a late HSV gene product. HSV caused a reduction of fibroblast cell surface HLA class I antigen, at least in part due to reduction of synthesis of heavy chain-f32 microglobulin heterodimers. These results suggest that HSV-induced blockade ofantigen presentation by cutaneous cells to CD8+ CTL may be a mechanism by which HSV limits or evades the immune response of the host. (J. Clin. Invest. 1993. 91:961-968.) Key words: HLA A2 e
Journal of General Virology, 1987
Herpes simplex virus type 1 (HSV-1) infection of human fibroblast cells grown in culture induces reorganization of the cytoskeleton fibrillar structures. Normal transport and insertion of HSV glycoproteins into the plasma membrane of the cells depend on the integrity of the microtubules. The natural host cells for HSV are epithelial cells, and an epithelial cell line established from rat palate was used in the present study. The effect of virus on the structure of the intermediate filaments and especially on the keratin proteins was studied. Two-dimensional gel electrophoresis of total cell extracts identified in uninfected cells two major acidic keratin proteins with apparent molecular weights of 44000 (44K) and 48K (pI 5.45 to 5.30, 5.50 to 5.35). A new keratin protein of46K (pI 5.40 to 5.25) appeared in infected cells between 8 h and 12 h post-infection. Pulse chase experiments identified the 46K protein as a processed form of the 48K keratin component, which was also cleaved in uninfected cells grown in the presence of cycloheximide. Partial proteolysis of the 46K and 48K keratins with Staphylococcus aureus V8 protease showed that the 48K and the 46K proteins differed in only one oligopeptide. The significance of the changed keratin composition of HSVinfected cells is discussed.
Journal of Virology, 2006
Sulfated Heparan Sulfate as O Role for 3http://jvi.asm.org/content/80/18/8970 Updated information and services can be found at: These include: REFERENCES http://jvi.asm.org/content/80/18/8970#ref-list-1 at: This article cites 71 articles, 35 of which can be accessed free CONTENT ALERTS more» articles cite this article), Receive: RSS Feeds, eTOCs, free email alerts (when new http://journals.asm.org/site/misc/reprints.xhtml Information about commercial reprint orders: http://journals.asm.org/site/subscriptions/ To subscribe to to another ASM Journal go to: on February 26, 2014 by guest http://jvi.asm.org/ Downloaded from on February 26, 2014 by guest
Journal of General Virology, 1986
Cultured human fibroblasts showed a typical fibriUar organization of microtubules in immunofluorescence, including the vimentin type of intermediate filament as well as actin-containing microfilaments. During infection with herpes simplex virus type 1 (HSV-1), the vimentin organization was maintained whereas actin, myosin and tubulin showed a progressive association with the viral glycoproteins within juxtanuclear structures. These structures could also be revealed with fluorochrome-coupled wheat germ agglutinin. Disruption of the microtubules by demecolcine treatment or their stabilization by taxol treatment did not prevent the aggregation of viral proteins in the cytoplasm. Taxol stabilization of the microtubules allowed the juxtanuclear accumulation of the glycoproteins in HSV-infected cells whereas treatment with demecolcine led to an accumulation of the glycoproteins either in small vesicles in the cytoplasm or in the focal adhesion areas of the cells. Production of infectious intracellular virus particles was reduced in cells treated with demecolcine or with taxol before and during infection. The results of this study indicate that the normal intracellular transport and distribution of the HSV glycoproteins and the formation of infectious virus are dependent on the presence of intact microtubules.
Expanding the role of 3-O sulfated heparan sulfate in herpes simplex virus type-1 entry
Virology, 2010
Heparan sulfate (HS) proteoglycans are commonly exploited by multiple viruses for initial attachment to host cells. Herpes simplex virus-1 (HSV-1) is unique because it can use HS for both attachment and penetration, provided specific binding sites for HSV-1 envelope glycoprotein gD are present. The interaction with gD is mediated by specific HS moieties or 3-O sulfated HS (3-OS HS), which are generated by all but one of the seven isoforms of 3-O sulfotransferases (3-OSTs). Here we demonstrate that several common experimental cell lines express unique sets of 3-OST isoforms. While the isoforms 3-OST-3,-5 and-6 were most commonly expressed, isoforms 3-OST-2 and-4 were undetectable in the cell lines examined. Since most cell lines expressed multiple 3-OST isoforms, we addressed the significance of 3-OS HS in HSV-1 entry by down-regulating 2-O-sulfation, a prerequisite for 3-OS HS formation, by knocking down 2-OST expression by RNA interference (RNAi). 2-OST knockdown was verified by reverse-transcriptase PCR and Western blot analysis, while 3-OS HS knockdown was verified by immunofluorescence. Cells showed a significant decrease in viral entry, suggesting an important role for 3-OS HS. Implicating 3-OS HS further, cells knocked down for 2-OST expression also demonstrated decreased cell-cell fusion when cocultivated with effector cells transfected with HSV-1 glycoproteins. Our findings suggest that 3-OS HS may play an important role in HSV-1 entry into many different cell lines.
Journal of General Virology, 1991
In previous studies, the herpes simplex virus type 1 (HSV-1) mutant, in1814, which lacks the transinducing function of Vmw65, did not replicate in the trigeminal ganglia of mice following corneal inoculation but did establish a reactivatable latent infection in the ganglia 12 to 24 h after ocular infection. Since in1814 did not replicate in vivo, the molecular events during the establishment phase of latent HSV-1 infection could be characterized without the complications of concurrent productive viral infection. In comparison to parental HSV-1 strain 17 ÷, the expression of viral immediate early (IE), early and late genes and the levels of viral DNA in the trigeminal ganglia of mice following in1814 infection were greatly reduced.