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Susceptibility of Human Female Primary Genital Epithelial Cells to Herpes Simplex Virus, Type-2 and the Effect of TLR3 Ligand and Sex Hormones on

Our site uses cookies to improve your experience. Methods: Human corneal epithelial cells (HCE) were infected with HSV-1 at a multiplicity of infection of 5. Previously, it was shown that an attenuated Herpes simplex virus vector (hrR3) transduced numerous cell types in the anterior and posterior segments of monkey eyes, but this was accompanied by inflammation. Epithelial cells, the portal of primary HSV-1 infection, support entry via low pH endocytosis mechanisms. Holmes. Basolateral infection and apical infection after the Ca(2+) switch required an intact microtubule network for genome targeting to the nucleus. Flow cytometry and immunocytochemistry were used to determine expression of three cellular receptors–nectin-1, herpesvirus entry mediator (HVEM), and paired immunoglobulin-like 2 receptor alpha (PILR-a).

LTKCRE and PGFPTK co-infected HLECs, but not RPECs, expressed high levels of the HSV-tk protein. F. Sparling, P. This mutation did not affect accumulation at cell junctions or cell-to-cell spread. There is no effective vaccine. Plaque formation is then widely analyzed, e.g., to determine the effect of mutation in virus-encoded genes on overall transmission efficiency, to identify distinct requirements for specific virus proteins during cell-to-cell transmission compared to those in extracellular entry (7–11), to analyze receptor usage and relocalization at cell-to-cell junctions (12), and to demonstrate host responses and interactions with incoming genomes (13), among many other types of investigation. Lemon, W.

E. Stamm, P. Piot, and J. The mice were anesthetized before inoculation by ether (Merck) inhalation and examined with the aid of a slit-lamp biomicroscope (SL 5 model; Kowa Co., Ltd., Nagoya, Japan) to exclude any animals with corneal trauma. Thus, unlike PRV, gE-deleted HSV-1 viruses have a retrograde spread defect in vivo. Sexually Transmitted Diseases. New York McGraw-Hill.

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K., P. F. The favored hypothesis is molecular mimicry in which virus and host proteins share short epitopes. A. Mardh, S. The growth medium was a mixture of Ham’s F12 and Dulbecco’s modified Eagle’s medium (1:2), supplemented with 0.5 μg/ml hydrocortisone, 10 ng/ml epidermal growth factor, 10% fetal calf serum, 2mmol/liter l-glutamine, 10 mmol/liter HEPES, 1 mmol/liter sodium pyruvate, 10−10 mol/liter cholera toxin, 5 μg/ml insulin, 5 μg/ml transferrin, and 15 × 10−4 mg/ml 3,3′,5′-triiodo-l-thyronine. Lemon, W.

E. Stamm, P. Piot, and J. N. Wasserheit. The experimental protocol including collection of discarded tissue had full University of California, Davis, Institutional Review Board (IRB) approval. ALDH1A can be expressed by LN stromal cells [29], [30] and ALDH1A expressing-RA-producing DCs are present in skin, lung and in their draining LNs [31].

1999. This redistribution was specifically to lateral cell surfaces, not to apical surfaces, and did not require the assembly of enveloped virions in the TGN. 2004 Report on the global HIVAIDS Epidemic: 4th global report [Internet]. (West Grove, Pa.). Joint United Nations Programme on HIV/AIDS (UNAIDS). Stable silencing was performed by means of lentiviruses encoding shRNA to β3 integrin (18, 25). The PCR product contained all of the 5′ coding sequences of gE but was truncated after residue 462, so that 16 residues of the gE CT domain remained.

21 Briefly, media were aspirated, and sorbitol (1.5 moles/L) in MEM was added to the cells and incubated for 1 hour at ambient temperature. Pixel intensity values from each profile were entered as data sets for determination of a Pearson correlation coefficient, as implemented at http://www.socscistatistics.com/tests/pearson. McQuillan, R. E. Johnson, A. J. Nahmias, S.

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Spear, P. At various times, the cells were scraped from dishes and pelleted and either the cells and medium or medium alone was frozen at −70°C and subsequently thawed and sonicated, and virus was then titered using Vero cells. Herpes simplex virus: receptors and ligands for cell entry. (B) HEC-1A cells were either infected with HSV-1 or coinfected with Ad(E1−)gE and Ad(E1−)gI. 6:401–410. Each column represents the average of triplicates ± standard deviation. HSV plaques formed on AdgEΔCT/gI-transduced HaCaT cells were significantly smaller than those formed on cells infected with control Ad vector, AdTet-trans, or on cells not infected with any Ad vector (Fig.

Stable mitochondrial transmembrane potential and caspase 9 activity were observed during HSV-1 infection (Fig. Numbers on the left are molecular sizes (in kilodaltons). Major, and S. E. Straus. Herpes simplex virus type 1 enters human epidermal keratinocytes, but not neurons, via a pH-dependent endocytic pathway. J Virol 2005.

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Huber, M. T., T. W. Wisner, N. R. The frontline boundary of moving cells is outlined (black) on one boundary. A.

Goldsmith, D. A. Rauch, R. J. We conclude that gE is required for efficient spread between epithelial cells and neurites, since NS-gEnull had a retrograde spread defect when added to HaCaT cells but not when the virus was added directly to neurites. Krummenacher, R. J.

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In: Gupta, S. Replication of gE CT domain mutants.To ensure that the defects observed in plaque formation by gE CT domain mutants were not due to defects in virus replication, we characterized the production of infectious virus and movement into extracellular compartments. Reproductive Immunology. 4B). 1999. In contrast, when nectin1 is uniformly distributed on the cell surface, nectin1-containing membranes remain in the bottom fractions of the gradients. All three of these glycoproteins appear to have been derived from a common ancestor (30).

Magnification ×500. Cytoplasmic membranes from HEp-2 cells infected with either HSV-1(F) or HSV-1 UL51Δ73-244 were fractionated on dextran gradients, and fractions were assayed by immunoblotting for VP5 (to identify virions) and for pUL51, pUL7, and gE (Fig. Laufer, and E. Gurpide. Culture of human endometrial cells under polarizing conditions. Differentiation 1990. 42:184–190.

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R. Distribution of gE CT domain mutants onto apical surfaces.HSV gE/gI binds the Fc domains of IgG on the surfaces of HSV-infected cells (2, 21, 22). Secretory component production by polarized epithelial cells from the human female reproductive tract. In other cell types, primarily highly transformed cell lines—e.g., Vero or R970 cells—that do not form extensive cell junctions, gE-gI does not affect cell-cell spread. 27:167–180. Classen-Linke, I., M. Kusche, R.

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