PDGF receptor-α does not promote HCMV entry into epithelial and endothelial cells but increased quantities stimulate entry by an abnormal pathway.-猫眼课题宝

DOI：

10.1371/journal.ppat.1002905

发表时间：

2012-09

期刊：

PLoS pathogens

影响因子：

6.7

通讯作者：

中科院分区：

医学1区

文献类型：

作者：

Vanarsdall AL;Wisner TW;Lei H;Kazlauskas A;Johnson DC

来源链接：

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3441672/?report=classic

Epidermal growth factor receptor (EGFR) and platelet-derived growth factor receptor-α (PDGFRα) were reported to mediate entry of HCMV, including HCMV lab strain AD169. AD169 cannot assemble gH/gL/UL128–131, a glycoprotein complex that is essential for HCMV entry into biologically important epithelial cells, endothelial cells, and monocyte-macrophages. Given this, it appeared incongruous that EGFR and PDGFRα play widespread roles in HCMV entry. Thus, we investigated whether PDGFRα and EGFR could promote entry of wild type HCMV strain TR. EGFR did not promote HCMV entry into any cell type. PDGFRα–transduction of epithelial and endothelial cells and several non-permissive cells markedly enhanced HCMV TR entry and surprisingly, promoted entry of HCMV mutants lacking gH/gL/UL128–131 into epithelial and endothelial cells. Entry of HCMV was not blocked by a panel of PDGFRα antibodies or the PDGFR ligand in fibroblasts, epithelial, or endothelial cells or by shRNA silencing of PDGFRα in epithelial cells. Moreover, HCMV glycoprotein induced cell-cell fusion was not increased when PDGFRα was expressed in cells. Together these results suggested that HCMV does not interact directly with PDGFRα. Instead, the enhanced entry produced by PDGFRα resulted from a novel entry pathway involving clathrin-independent, dynamin-dependent endocytosis of HCMV followed by low pH-independent fusion. When PDGFRα was expressed in cells, an HCMV lab strain escaped endosomes and tegument proteins reached the nucleus, but without PDGFRα virions were degraded. By contrast, wild type HCMV uses another pathway to enter epithelial cells involving macropinocytosis and low pH-dependent fusion, a pathway that lab strains (lacking gH/gL/UL128–131) cannot follow. Thus, PDGFRα does not act as a receptor for HCMV but increased PDGFRα alters cells, facilitating virus entry by an abnormal pathway. Given that PDGFRα increased infection of some cells to 90%, PDGFRα may be very useful in overcoming inefficient HCMV entry (even of lab strains) into the many difficult-to-infect cell types. Human cytomegalovirus (HCMV) causes substantial morbidity and mortality in immunocompromised patients and in developing infants. HCMV pathogenesis involves the capacity to infect many different cell types by multiple distinct entry pathways. Among the biologically important cell types infected in vivo are epithelial and endothelial cells. HCMV specifically requires the viral glycoprotein gH/gL/UL128–131 to enter these cells. Previous studies suggested that platelet derived growth factor receptor-α (PDGFRα) was important for HCMV entry into cells. We characterized whether PDGFRα was important for HCMV entry. Increased expression of PDGFRα in cells markedly augmented entry of wild type and gH/gL/UL128–131-mutant HCMV into epithelial and endothelial cells, however, other experiments showed that endogenous PDGFRα did not normally mediate HCMV entry into these cell types. Instead, the increased expression of PDGFRα increased HCMV entry by an abnormal pathway involving clathrin-independent endocytosis and low pH-independent fusion with endosomes. HCMV normally enters these cells by macropinocytosis and low pH-dependent fusion. Therefore, PDGFRα is not normally an HCMV entry mediator in these cells, but increased expression of PDGFRα can promote entry by a different pathway. PDGFRα transduction of cells may be very useful because many cells are poorly infected by HCMV and entry represents a major hurdle.