Platelet-derived growth factor-α receptor is the cellular receptor for human cytomegalovirus gHgLgO trimer

Kabanova, Anna; Marcandalli, Jessica; Zhou, Tongqing; Bianchi, Siro; Baxa, Ulrich; Tsybovsky, Yaroslav; Lilleri, Daniele; Silacci-Fregni, Chiara; Foglierini, Mathilde; Fernandez-Rodriguez, Blanca; Druz, Aliaksandr; Zhang, Baoshan; Geiger, Roger; Pagani, Massimiliano; Sallusto, Federica; Kwong, Peter D.; Corti, Davide; Lanzavecchia, Antonio; Perez, Laurent

doi:10.1038/nmicrobiol.2016.82

Cited by 163 publications

(258 citation statements)

References 39 publications

Supporting

Mentioning

236

Contrasting

Order By: Relevance

“…In all experiments, gO-deficient virus mutants were less impaired in infection of HECs than of HFFs. This incomplete dependency of cell-free infection on gO could either be a characteristic of endothelial cells, e.g., via nonspecific endocytic uptake, or it could indicate that gO has two distinct functions in entry: a direct influence on fusion, which is important for entry into all cell types (21), and a fibroblast specific function, e.g., receptor binding, as suggested by a recent publication (34). A detailed analysis of HCMV-specific parts of gO will help to answer this question.…”

Section: Discussionmentioning

confidence: 97%

See 1 more Smart Citation

Importance of Highly Conserved Peptide Sites of Human Cytomegalovirus gO for Formation of the gH/gL/gO Complex

Stegmann¹,

Abdellatif

Sampaio³

et al. 2017

J Virol

View full text Add to dashboard Cite

The glycoprotein O (gO) is betaherpesvirus specific. Together with the viral glycoproteins H and L, gO forms a covalent trimeric complex that is part of the viral envelope. This trimer is crucial for cell-free infectivity of human cytomegalovirus (HCMV) but dispensable for cell-associated spread. We hypothesized that the amino acids that are conserved among gOs of different cytomegaloviruses are important for the formation of the trimeric complex and hence for efficient virus spread. In a mutational approach, nine peptide sites, containing all 13 highly conserved amino acids, were analyzed in the context of HCMV strain TB40-BAC4 with regard to infection efficiency and formation of the gH/gL/gO complex. Mutation of amino acids (aa) 181 to 186 or aa 193 to 198 resulted in the loss of the trimer and a complete small-plaque phenotype, whereas mutation of aa 108 or aa 249 to 254 caused an intermediate phenotype. While individual mutations of the five conserved cysteines had little impact, their relevance was revealed in a combined mutation, which abrogated both complex formation and cell-free infectivity. C343 was unique, as it was sufficient and necessary for covalent binding of gO to gH/gL. Remarkably, however, C218 together with C167 rescued infectivity in the absence of detectable covalent complex formation. We conclude that all highly conserved amino acids contribute to the function of gO to some extent but that aa 181 to 198 and cysteines 343, 218, and 167 are particularly relevant. Surprisingly, covalent binding of gO to gH/gL is required neither for its incorporation into virions nor for proper function in cell-free infection.IMPORTANCE Like all herpesviruses, the widespread human pathogen HCMV depends on glycoproteins gB, gH, and gL for entry into target cells. Additionally, gH and gL have to bind gO in a trimeric complex for efficient cell-free infection. Homologs of gO are shared by all cytomegaloviruses, with 13 amino acids being highly conserved. In a mutational approach we analyzed these amino acids to elucidate their role in the function of gO. All conserved amino acids contributed either to formation of the trimeric complex or to cell-free infection. Notably, these two phenotypes were not inevitably linked as the mutation of a charged cluster in the center of gO abrogated cell-free infection while trimeric complexes were still being formed. Cysteine 343 was essential for covalent binding of gO to gH/gL; however, noncovalent complex formation in the absence of cysteine 343 also allowed for cell-free infectivity.

show abstract

Section: Discussionmentioning

confidence: 97%

“…Immunization of mice with gH/gL/gO leads to full protection from otherwise lethal MCMV infection (33). For HCMV it was recently demonstrated that gH/gL/gO interacts with the cellular receptor platelet-derived growth factor receptor alpha (PDGFR-␣) (34). Taken together, these findings highlight that gO is a promising target for future vaccination strategies.…”

mentioning

confidence: 95%

Importance of Highly Conserved Peptide Sites of Human Cytomegalovirus gO for Formation of the gH/gL/gO Complex

Stegmann¹,

Abdellatif

Sampaio³

et al. 2017

J Virol

View full text Add to dashboard Cite

show abstract

“…In more detail, gH/gL may be complexed either with pUL128L, giving rise to the PC gH/gL/ pUL128L, or UL74-encoded gO, thus forming the trimeric complex gH/gL/gO, which binds to platelet-derived growth factor receptor a and mediates HCMV entry into HELFs [57][58][59]. Thus, gH/gL/gO and PC were considered to represent two mutually exclusive cell entry complexes, as suggested by mass spectrometry and mutagenesis analysis [60].…”

Section: Genetic Determinants Of Endothelial Cells and Leukocyte Tropismmentioning

confidence: 99%

“…However, gO and the UL128-131 gene products were reported to compete for binding of gH/gL, thus influencing the ratio of gH/gL/ gO to gH/gL/UL128-131 within virions as well as the HCMV cell tropism [61,62]. As a result, gH/gL/gO was also found to be required for entry into epithelial cells [57,63,64].…”

Section: Genetic Determinants Of Endothelial Cells and Leukocyte Tropismmentioning

confidence: 99%

The pentameric complex of human Cytomegalovirus: cell tropism, virus dissemination, immune response and vaccine development

Gerna

Revello

Baldanti

et al. 2017

Journal of General Virology

Self Cite

View full text Add to dashboard Cite

Between the 1980s and 1990s, three assays were developed for diagnosis of human cytomegalovirus (HCMV) infections: leuko (L)-antigenemia, L-viremia and L-DNAemia, detecting viral protein pp65, infectious virus and viral DNA, respectively, in circulating leukocytes Repeated initial attempts to reproduce the three assays in vitro using laboratory-adapted strains and infected cell cultures were consistently unsuccessful. Results were totally reversed when wild-type HCMV strains were used to infect either fibroblasts or endothelial cells. Careful analysis and sequencing of plaque-purified viruses from recent clinical isolates drew attention to the ULb¢ region of the HCMV genome. Using bacterial artificial chromosome technology, it was shown by both gain-of-function and loss-of-function experiments that UL131-128 genes are indispensable for virus growth in endothelial cells and virus transfer to leukocytes. In addition, a number of clinical isolates passaged in human fibroblasts had lost both properties (leuko-tropism and endothelial cell-tropism) when displaying a mutation in the UL131-128 locus

show abstract

“…Fibroblast viral entry requires gB and a trimeric complex of glycoproteins gH, gL, and gO, whereas viral entry into endothelial, epithelial, and dendritic cells requires an additional pentameric complex (pentamer) of gH, gL, UL128, UL130, and UL131 (24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34). Consequently, antibodies directed against gB, gH/gL, or gO impair viral entry into fibroblasts and endothelial and epithelial cells, whereas antibodies that specifically target the pentamer selectively block viral entry into epithelial and endothelial cells (28,33,(35)(36)(37). Following natural infection, the latter activity is dominant, as the serum neutralizing titers measured with epithelial cells are significantly higher than those measured using fibroblasts (38)(39)(40)(41).…”

mentioning

confidence: 99%

Impact of Antibodies and Strain Polymorphisms on Cytomegalovirus Entry and Spread in Fibroblasts and Epithelial Cells

Cui

Freed

Wang

et al. 2017

J Virol

View full text Add to dashboard Cite

Cytomegalovirus (CMV) entry into fibroblasts differs from entry into epithelial cells. CMV also spreads cell to cell and can induce syncytia. To gain insights into these processes, 27 antibodies targeting epitopes in CMV virion glycoprotein complexes, including glycoprotein B (gB), gH/gL, and the pentamer, were evaluated for their effects on viral entry and spread. No antibodies inhibited CMV spread in fibroblasts, including those with potent neutralizing activity against fibroblast entry, while all antibodies that neutralized epithelial cell entry also inhibited spread in epithelial cells and a correlation existed between the potencies of these two activities. This suggests that exposure of virions to the cell culture medium is obligatory during spread in epithelial cells but not in fibroblasts. In fibroblasts, the formation of syncytiumlike structures was impaired not only by antibodies to gB or gH/gL but also by antibodies to the pentamer, suggesting a potential role for the pentamer in promoting fibroblast fusion. Four antibodies reacted with linear epitopes near the N terminus of gH, exhibited strain specificity, and neutralized both epithelial cell and fibroblast entry. Five other antibodies recognized conformational epitopes in gH/gL and neutralized both fibroblast and epithelial cell entry. That these antibodies were strain specific for neutralizing fibroblast but not epithelial cell entry suggests that polymorphisms external to certain gH/gL epitopes may influence antibody neutralization during fibroblast but not epithelial cell entry. These findings may have implications for elucidating the mechanisms of CMV entry, spread, and antibody evasion and may assist in determining which antibodies may be most efficacious following active immunization or passive administration.IMPORTANCE Cytomegalovirus (CMV) is a significant cause of birth defects among newborns infected in utero and morbidity and mortality in transplant and AIDS patients. Monoclonal antibodies and vaccines targeting humoral responses are under development for prophylactic or therapeutic use. The findings reported here (i) confirm that cell-to-cell spread of CMV is sensitive to antibody inhibition in epithelial cells but not fibroblasts, (ii) demonstrate that antibodies can restrict the formation in vitro of syncytiumlike structures that resemble syncytial cytomegalic cells that are associated with CMV disease in vivo, and (iii) reveal that neutralization of CMV by antibodies to certain epitopes in gH or gH/gL is both strain and cell type dependent and can be governed by polymorphisms in sequences external to the epitopes. These findings serve to elucidate the mechanisms of CMV entry, spread, and antibody evasion and may have important implications for the development of CMV vaccines and immunotherapeutics.

show abstract

Platelet-derived growth factor-α receptor is the cellular receptor for human cytomegalovirus gHgLgO trimer

Cited by 163 publications

References 39 publications

Importance of Highly Conserved Peptide Sites of Human Cytomegalovirus gO for Formation of the gH/gL/gO Complex

Importance of Highly Conserved Peptide Sites of Human Cytomegalovirus gO for Formation of the gH/gL/gO Complex

The pentameric complex of human Cytomegalovirus: cell tropism, virus dissemination, immune response and vaccine development

Impact of Antibodies and Strain Polymorphisms on Cytomegalovirus Entry and Spread in Fibroblasts and Epithelial Cells

Contact Info

Product

Resources

About