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1.
FIG. 8.

FIG. 8. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

HCMV and RhCMV gL is unable to complement heterologous gL-null viruses. Telo-RF and HFF cells were mock transduced or were transduced with retroviruses expressing either RhCMV or HCMV gL and subsequently infected with 100 PFU of the viruses indicated. At 14 days postinfection, photomicrographs were obtained.

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.
2.
FIG. 7.

FIG. 7. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

Rescue of HCMVΔgL infectivity in HFFs expressing HCMV gL. (A) HFFs were mock transduced or transduced twice with retrovirus expressing HCMV gL. Expression of gL was indirectly confirmed by expression of GFP from the IRES:GFP cassette. (B) HCMVΔgL replicates in cells expressing HCMV gL. HFF and HFF:HgL cells were infected with HCMVΔgL, and photomicrographs were obtained 18 days after infection.

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.
3.
FIG. 5.

FIG. 5. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

gB expression and virion morphogenesis in cells infected with of RhCMVΔgL is similar to RhCMV. (A) Telo-RF cells were infected with 2.5 PFU of RhCMV or RhCMVΔgL/cell. At 72 h postinfection, cell lysates were prepared, and gB was detected by Western blotting. (B) Telo-RF cells were infected with either RhCMV (upper panels) or RhCMVΔgL (lower panels) at an MOI of 3. At 5 days postinfection the cells were fixed, and transmission electron microscopy was performed. RhCMV and RhCMVΔgL virions are detected in the cytoplasm (left panels) and outside the cell (right panels).

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.
4.
FIG. 6.

FIG. 6. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

The infectivity of RhCMVΔgL can be rescued by treatment with PEG. RhCMVΔgL passaged in noncomplementing Telo-RF cells was used to infect Telo-RF cells or Telo-RF cells expressing RhCMV gL for 1 h at 37°C. The virus inoculum was removed, and the cells were washed three times in serum-free DMEM (left panels) or DMEM containing PEG (center panels). At 5 days postinfection, the supernatant from the PEG-treated cells was transferred to monolayers of complementing Telo-RF:RhgL cells (right panels).

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.
5.
FIG. 3.

FIG. 3. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

Expression of GFP and RhCMV gL in Telo-RF:RhgL cells. (A) Telo-RF cells were mock transduced or transduced twice with retrovirus expressing RhCMV gL and GFP. RhCMV gL expression was indirectly measured by expression of GFP from the IRES following the gL gene in the retroviral transcript. Images of GFP expression were taken 6 months after transduction. (B) gL expression in Telo-RF:RhgL cells. Telo-RF and Telo-RF:RhgL cells were fixed and stained for gL using rabbit polyclonal antibody raised against gL peptide, followed by anti-rabbit Texas Red-conjugated antibody. Cells were costained with DAPI to visualize nuclei.

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.
6.
FIG. 1.

FIG. 1. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

Naturally infected rhesus monkeys do not produce detectable levels of antibody to RhCMV gL. Antibody levels to RhCMV gB, pp65.2, and gL were measured by LIPS assay with 17 RhCMV-seronegative and 11 RhCMV-seropositive animals. The data are presented as the amount of light units (LU) produced after immunoprecipitation. The long horizontal bars indicate the mean luminescence, and the short horizontal bars represent the standard deviations. Serum obtained from preimmune rabbits or rabbits immunized with gL peptides were included as a negative and positive controls, respectively, for the gL LIPS assay.

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.
7.
FIG. 4.

FIG. 4. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

RhCMVΔgL forms plaques only in cells expressing RhCMV gL. (A) Telo-RF cells expressing RhCMV gL were transfected with RhCMVΔgL BAC DNA. When the cells showed 90% CPE, the supernatant was harvested, clarified by low-speed centrifugation, and then added to either nontransduced Telo-RF cells (RF, upper panels) or Telo-RF cells expressing RhCMV gL (Telo-RF:RhgL, lower panels). At 4 and 10 days postinfection (dpi), photomicrographs of cell monolayers were obtained. (B) Replication of RhCMVΔgL in complementing cells. Telo-RF cells stably transduced to express RhCMV gL were infected with RhCMV or RhCMVΔgL at an MOI of 2.5 PFU/cell. At the times indicated, supernatant virus was collected and assayed for infectious virus. The data presented are the averages of two experiments, and error bars indicate the standard deviations. (C) Expression of gB and gL in Telo-RF cells infected with RhCMV or RhCMVΔgL. Telo-RF cells were infected at an MOI of 1 PFU/cell. At 3 days after infection, the cells were fixed and stained for either gB or gL as indicated.

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.
8.
FIG. 2.

FIG. 2. From: Rhesus and Human Cytomegalovirus Glycoprotein L Are Required for Infection and Cell-to-Cell Spread of Virus but Cannot Complement Each Other .

Map and Southern blot of the region of RhCMV with gL deleted. (A) Map of the RhCMV genome in the region containing gL and adjacent genes. Nucleotide positions of the BlpI sites and the gL open reading frame (ORF) are indicated. The white arrow indicates the orientation and position of the gL ORF. The gray arrows indicate the positions and orientation of the adjacent ORFs. The dotted line denotes the location of the probe used for Southern blot analysis. (B) Map of the RhCMVΔgL genome in the region of the gL deletion. The nucleotide positions of the beginning and end of the deletion are noted. Loss of the majority of the gL ORF results in loss of a single BlpI site. (C) Southern blot of RhCMV BAC DNA, RhCMV virion DNA, RhCMVΔgL BAC DNA, and RhCMVΔgL virion DNA digested with BlpI and hybridized with a radiolabeled probe specific for the region overlapping RhCMV gL and the upstream neighboring gene. The probe hybridizes to a 6.1-kb BlpI fragment in RhCMV DNA and a 9.2-kb BlpI fragment in RhCMVΔgL DNA. Size markers are in kilobase pairs.

J. Jason Bowman, et al. J Virol. 2011 March;85(5):2089-2099.

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