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The PDGF ligand (PDGF-AA) and PDGFRα antibodies do not reduce HCMV infection of epithelial, endothelial cells, and fibroblasts.

By Adam L. Vanarsdall (307887), Todd W. Wisner (307889), Hetian Lei (307891), Andrius Kazlauskas (307893) and David C. Johnson (307895)

Abstract

<p>(<b>A</b>) ARPE-19, HUVECs, or foreskin fibroblasts (HFF) were pre-treated with PDGF-AA ligand at various concentrations at 4<b>°</b>C for 30 min then HCMV TR (10 IU/cell, 1 IU/cell for fibroblasts) was added to the cells in the presence of PDGF-AA at 4<b>°</b>C for an additional 30 min. The cells were shifted to 37<b>°</b>C for 4 hr then washed to remove the virus inoculum and fresh growth media containing the same concentration of PDGF-AA was added. After an additional 24 hr, the cells were fixed, permeabilized, and stained for HCMV IE-86. Relative entry was characterized by comparing the number of IE-86 positive cells in three separate fields with other dishes of cells not treated with PDGFR-AA. Error bars indicate the standard deviation. <b>B</b>) ARPE-19 cells, <b>C</b>) HUVE cells or <b>D</b>) human foreskin fibroblasts were incubated with various doses of PDGFRα-specific antibodies for 1 hr at 4°C then incubated with HCMV TR (10 IU/cell, 1 PFU/cell for fibroblasts) in the presence of these antibodies for an additional 1 hr at 4°C. The cells were then shifted to 37°C for 4 hr, washed to remove the virus inoculum then incubated at 37°C for 24 hr in the presence of antibodies. The cells were then stained for IE-86. Relative entry refers to the frequency of IE-86 positive cells compared to cells not treated with antibodies and was determined from at least three separate fields. Error bars indicate the standard deviation.</p

Topics: Biological Sciences, microbiology, Molecular cell biology, pdgf, ligand, antibodies, hcmv, endothelial
Year: 2013
DOI identifier: 10.1371/journal.ppat.1002905.g005
OAI identifier: oai:figshare.com:article/248633
Provided by: FigShare
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