Glycoprotein B of Human Cytomegalovirus Promotes Virion Penetration into Cells, Transmission of Infection from Cell to Cell, and Fusion of Infected Cells

Navarro, David; Paz, Pedro; Tugizov, Sharof M.; Topp, K; Vail, J La; Pereira, Lenore

doi:10.1006/viro.1993.1575

Cited by 214 publications

(210 citation statements)

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“…Analysis of infection showed that the apical membrane of polarized RPE cells was more susceptible to infection by HCMV than was the basal membrane and that neutralizing monoclonal antibodies (MAbs) to glycoprotein B (gB) blocked virus penetration of the apical membrane. In contrast to our previous findings with HCMV-infected unpolarized human foreskin fibroblast (HFF) cells (Navarro et al, 1993), neutralizing MAbs to gB failed to inhibit plaque development in polarized RPE cells, which suggested that progeny virions were transmitted from cell to cell before tight junctions were altered and were shielded from neutralization.…”

Section: Introductioncontrasting

confidence: 99%

“…Stock virus preparations of HCMV strain AD169 were produced and titred for infectivity in HFF cells, using the rapid infectivity assay (Andreoni et al, 1989;Navarro et aL, 1993). Herpes simplex virus type 1 (HSV-I) strain F was propagated and titred in Vero cells as described previously (Ejercito et al, 1968).…”

Section: Cells Culture Media Viruses and Propagation Of Cells On Pementioning

confidence: 99%

“…Properties of neutralizing MAbs to HCMV gB (UL55), mapping of their epitopes and analysis of neutralization by blocking fusion of the virion envelope with the plasma membrane have been published (Basgoz et al, 1992;Navarro et al, 1993;Pereira et al, 1984Pereira et al, , 1991Qadri et al, 1992). Immunoglobulin (Ig) was purified from murine ascites fuids by Protein A affinity chromatography according to the manufacturer's instructions (Affi-Gel protein A MAPS II Kit; Bio-Rad).…”

Section: Cells Culture Media Viruses and Propagation Of Cells On Pementioning

confidence: 99%

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Role of apical and basolateral membranes in replication of human cytomegalovirus in polarized retinal pigment epithelial cells

Tugizov

Maidji

Pereira

1996

Journal of General Virology

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Human retinal pigment epithelial (RPE) cells, which are permissive for human cytomegalovirus (HCMV) replication, were used to evaluate virus infection from apical and basolateral membranes of polarized cells. Tests of HCMV infectivity showed that the apical membrane was 20-30-fold more susceptible to infection than the basolateral membrane; in contrast, both membranes were equally susceptible to infection by herpes simplex virus type 1 (HSV-1). Neutralizing monoclonal antibodies (MAbs) to HCMV glycoprotein B (gB) blocked penetration of virions into polarized RPE cells. This indicated that gB has a function in fusion of the virion envelope with the apical membrane of these cells, as it has with the cell membrane of unpolarized human fibroblasts. In contrast to HSV-l-infected RPE cells, the paracellular permeability of polarized RPE cells changed slowly following infection with HCMV. Confocal microscopy examination of HCMV-infected RPE cells revealed that the pattern of ZO-1 staining was altered at late times. Addition ofgB-specific neutralizing MAbs to the apical and basolateral membranes of HCMV-infected RPE cells failed to inhibit plaque development; this indicated that progeny virions infect adjacent cells before disassembly of tight junctions and are sequestered from neutralization during spread across lateral cell membranes. The finding that progeny HCMV virions cross lateral cell membranes, which differ substantially in protein composition from apical membranes, suggests that polarized RPE cells contain multiple receptors for HCMV.

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Section: Introductioncontrasting

confidence: 99%

Section: Cells Culture Media Viruses and Propagation Of Cells On Pementioning

confidence: 99%

Section: Cells Culture Media Viruses and Propagation Of Cells On Pementioning

confidence: 99%

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Role of apical and basolateral membranes in replication of human cytomegalovirus in polarized retinal pigment epithelial cells

Tugizov

Maidji

Pereira

1996

Journal of General Virology

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“…HCMV glycoprotein B (gB) gene is considered to be a multifunctional envelope component responsible for virion entry, cell to cell spread, syncytium formation 18 and is the major target for neutralizing antibodies 4,5 . Based on sequence variation of the gB gene, HCMV strains can be classified into four gB genotypes 5 and recently, two new genotypes (gB6 and gB7) have been reported by TRINCADO et al 27 .…”

Section: Introductionmentioning

confidence: 99%

Determination of human cytomegalovirus genetic diversity in different patient populations in Costa Rica

Ahumada-Ruiz

Taylor-Castillo

Visoná

et al. 2004

Rev. Inst. Med. trop. S. Paulo

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SUMMARYSeroprevalence of HCMV in Costa Rica is greater than 95% in adults; primary infections occur early in life and is the most frequent congenital infection in newborns. The objectives of this study were to determine the genetic variability and genotypes of HCMV gB gene in Costa Rica. Samples were collected from alcoholics, pregnant women, blood donors, AIDS patients, hematologyoncology (HO) children and HCMV isolates from neonates with cytomegalic inclusion disease. A semi-nested PCR system was used to obtain a product of 293-296 bp of the gB gene to be analyzed by Single Stranded Conformational Polymorphism (SSCP) and sequencing to determine the genetic polymorphic pattern and genotypes, respectively. AIDS patients showed the highest polymorphic diversity with 14 different patterns while fifty-six percent of HO children samples showed the same polymorphic pattern, suggesting in this group a possible nosocomial infection. In neonates three genotypes (gB1, gB2 and gB3), were determined while AIDS patients and blood donors only showed one (gB2). Of all samples analyzed only genotypes gB1, 2 and 3 were determined, genotype gB2 was the most frequent (73%) and mixed infections were not detected. The results of the study indicate that SSCP could be an important tool to detect HCMV intra-hospital infections and suggests a need to include additional study populations to better determine the genotype diversity and prevalence.

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“…The surface subunit gp116 and the transmembrane subunit gp55 are disufide-bonded. gB plays an important role in virus infectivity and cell-to-cell spread, and is a major target for antibody-mediated immunity (Navarro et al, 1993 ;Britt et al, 1990 ;Utz et al, 1989). The linear epitopes AD1 (located on gp 55) and AD2 (located at the N terminus of gp116) bind virus-neutralizing antibodies (Britt & Mach, 1996 ;Spaete et al, 1988).…”

Section: Introductionmentioning

confidence: 99%

Variation within the glycoprotein B gene of human cytomegalovirus is due to homologous recombination.

Haberland¹,

Meyer-K√∂nig²,

Hufert³

1999

Journal of General Virology

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Human cytomegalovirus (HCMV) strains can be classified into different glycoprotein B (gB) genotypes. In a previous study, frequent intragenic variation of the gB gene was shown. The aim of this study was to analyse whether gB variation was due to homologous recombination. The gB gene of DNA extracts derived from the peripheral blood leukocytes of 14 immunosuppressed patients was amplified by PCR and cloned. Three variable sites of gB were analysed by restriction fragment analysis and DNA sequencing and compared with published prototypic strains. In three patients doubly infected with two distinct HCMV gB strains, prototypic (60-85 %) and nonprototypic recombinant strains (5-40 %) were detected. To demonstrate that homologous recombination is driving HCMV gB variability, cells were coinfected with plaque-purified prototypic gB strains and recombinant gB genes were selectively amplified by PCR. gB recombinants were detected after 15 days of coculture and cross-over sites were determined by sequencing. These data indicate that homologous recombination contributes to the variability of the gB gene in vitro and in vivo.

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Glycoprotein B of Human Cytomegalovirus Promotes Virion Penetration into Cells, Transmission of Infection from Cell to Cell, and Fusion of Infected Cells

Cited by 214 publications

References 0 publications

Role of apical and basolateral membranes in replication of human cytomegalovirus in polarized retinal pigment epithelial cells

Role of apical and basolateral membranes in replication of human cytomegalovirus in polarized retinal pigment epithelial cells

Determination of human cytomegalovirus genetic diversity in different patient populations in Costa Rica

Variation within the glycoprotein B gene of human cytomegalovirus is due to homologous recombination.

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