The Ebola-Glycoprotein Modulates the Function of Natural Killer Cells

Edri, Avishay; Shemesh, Avishai; Iraqi, Muhammed; Matalon, Omri; Brusilovsky, Michael; Hadad, Uzi; Radinsky, Olga; Gershoni-Yahalom, Orly; Dye, John M.; Mandelboim, Ofer; Barda‐Saad, Mira; Lobel, Leslie; Porgador, Angel

doi:10.3389/fimmu.2018.01428

Cited by 22 publications

(30 citation statements)

References 71 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Here, we observed a lower binding of NKG2D-Fc chimeric soluble receptors to EBOV-GP transfected HEK 293 cells corresponding to reduced expression of MICA/B (Fig. 6b, bottom), as has been reported before (115). Interestingly, we reported previously for Vaccinia virus (VV)-infected HeLa cells that the cumulative expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3, and ULBP4) was identical to that in noninfected cells, as detected with NKG2D-Fc (59,141,142).…”

Section: Ebov-gp Protects Hek-293 Cells From Lysis By Polyclonal Nk Csupporting

confidence: 88%

“…6a). In addition, we observed a lower binding of NKG2D-Fc chimeric soluble receptors to EBOV-GP transfected HEK 293 cells ( Fig.6b), which indicates a reduction of its ligands MIC-A/B, as it has been shown before (115). Altogether, these data suggest that a by EBOV-GP reduced stimulation of NKG2D, together with the recruitment of Siglec-ligands on the surface of infected cells, may lead to a suppressive state of effector cells (Fig.…”

Section: Psgl-1 Cd24 Cd44 Dr3 and Ebola-gp Are Receptors For Selecsupporting

confidence: 80%

See 1 more Smart Citation

Activating NK- receptors, homing selectins and inhibitory Siglecs recognize EBOLA-GP and HPV-L1NK

Jarahian

Marstaller

Wurmbäck

et al. 2020

Preprint

View full text Add to dashboard Cite

The Ebola virus glycoprotein (EBOV)-GP is extensively glycosylated. Its expression induces a physical alteration of surface adhesion molecules, which causes cell rounding and detachment of the infected cells. This phenomenon likely plays a crucial role in viral pathogenicity. In this study, we show that such morphological changes are cell line-dependent as well as dependent on the surface proteins that interact with EBOV-GP in cis and trans. We have generated data showing that natural killer (NK) cell receptors (NCRs: NKp44 and NKp46), selectins (CD62E/P/L) and inhibitory Siglecs function as receptors for Ebola-GP and human papilloma virus (HPV-L1). We used HEK293 cells transfected with Ebola-GP and recombinant fusion proteins containing the extracellular domain of each of these receptors linked to the Fc of human IgG1, which showed significant differences in their virus-binding behavior compared to HEK293 cells transfected with empty vector. Further, to demonstrate that EBOV-GP is a ligand for NKp44 and other NK-receptors, and to investigate their role in immune escape, we also used human HEK-293, HeLa- and hamster CHO-GP-transfectants. Our data show that the NK receptors NKp44 and NKp46 play a key role in recognizing EBOV (Ebolavirus) and strongly suggest that other inhibitory (Siglec-7, Siglec-5) and non-inhibitory homing receptors (P-Selectin, L-Selectin, E-Selectin, and DC-SIGNR/DC-SIGN) take part in the interaction with virus particles. In addition, we show that NKp44, and NKp46, Siglec-7, and -5, and P-, L-, E-selectins as well as of and DC-SIGNR/DC-SIGN bind to the artificial viral envelope of a lentiviral vector that contains EBOV-GP. Altogether we prove that NCRs and a range of other inhibitory and activating receptors can interact with viral envelope/capsid proteins and that such interaction could play an important role in the elimination of virus infected cells. Our findings could be used to develop new strategies for prevention and treatment of infections by these viruses.Author summaryThe innate immune system is able to recognize specifically certain virus components. Here we show that activating NK-cell receptors (NKp44, and NKp46) are involved in such interaction by using HEK293 and CHOK1 cells transfected with the Ebola virus glycoprotein (EBOV-GP) and by binding studies with purified EBOV-GP. In detail, we have found moderate to strong affinity of Siglecs (Siglec-7, and -5), selectins (P-, L-, E-Selectin) and DC-SIGNR/DC-SIGN to purified EBOV-GP, and to cells transfected with EBOV-GP as well as to the envelope of a lentiviral vector carrying the EBOV-GP. Our findings show that NKp44, and NKp46, Siglec-7, and -5, as well as P-and L-selectins have a strong affinity to EBOV-G.

show abstract

Section: Ebov-gp Protects Hek-293 Cells From Lysis By Polyclonal Nk Csupporting

confidence: 88%

Section: Psgl-1 Cd24 Cd44 Dr3 and Ebola-gp Are Receptors For Selecsupporting

confidence: 80%

Activating NK- receptors, homing selectins and inhibitory Siglecs recognize EBOLA-GP and HPV-L1NK

Jarahian

Marstaller

Wurmbäck

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…We and others have shown that expression of EBOV-GP shields cell membrane HLA-I from recognition by antibodies or by TCR (Ignatiev et al, 2000;Geisbert et al, 2003;Reed et al, 2004). We then showed that expression of EBOV-GP on target cells also shields ligands to activating NK receptors (Edri et al, 2018). The GP1 fragment of the GP was mediating this shielding since both trypsin and DTT treatments of the cells that remove the GP1 (enzymatically and chemically, respectively) abolished the shielding (Edri et al, 2018).…”

Section: N-glycosylation Of Mld and Gcd Domains Of Gp1 Is Involved Inmentioning

confidence: 82%

“…We then showed that expression of EBOV-GP on target cells also shields ligands to activating NK receptors (Edri et al, 2018). The GP1 fragment of the GP was mediating this shielding since both trypsin and DTT treatments of the cells that remove the GP1 (enzymatically and chemically, respectively) abolished the shielding (Edri et al, 2018). We now investigated whether the N-glycosylation of the GP1 is the major factor mediating this shielding.…”

Section: N-glycosylation Of Mld and Gcd Domains Of Gp1 Is Involved Inmentioning

confidence: 98%

“…Peptide-presenting HLA-I are the ligands for the T cell receptor but HLA-I also serve as ligands for NK inhibitory receptors (Kärre et al, 1986;Ljunggren and Kärre, 1990;Colonna and Samaridis, 1995;D'andrea et al, 1995;Wagtmann et al, 1995;Braud et al, 1998;Lanier, 1998;Davis et al, 1999;López-Botet and Bellón, 1999;Kim et al, 2005;Vivier et al, 2011). We previously investigated the effect of EBOV-GP on NK cell function (Edri et al, 2018). We showed that EBOV-GP also effectively shields ligands to activating NK receptors such as MHC class I polypeptide-related sequence A (MICA) and B7-H6; overall, we showed that EBOV-GP expression by target cells favors the axis reducing NK activation, while not perturbing the NK inhibition axis (Braiman et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

N-Glycans Mediate the Ebola Virus-GP1 Shielding of Ligands to Immune Receptors and Immune Evasion

Iraqi

Edri

Greenshpan

et al. 2020

Front. Cell. Infect. Microbiol.

Self Cite

View full text Add to dashboard Cite

The Ebola Virus (EBOV) glycoprotein (GP) sterically shields cell-membrane ligands to immune receptors such as human leukocyte antigen class-1 (HLA-I) and MHC class I polypeptide-related sequence A (MICA), thus mediating immunity evasion. It was suggested that the abundant N-glycosylation of the EBOV-GP is involved in this steric shielding. We aimed to characterize (i) the GP N-glycosylation sites contributing to the shielding, and (ii) the effect of mutating these sites on immune subversion by the EBOV-GP. The two highly glycosylated domains of GP are the mucin-like domain (MLD) and the glycan cap domain (GCD) with three and six N-glycosylation sites, respectively. We mutated the N-glycosylation sites either in MLD or in GCD or in both domains. We showed that the glycosylation sites in both the MLD and GCD domains contribute to the steric shielding. This was shown for the steric shielding of either HLA-I or MICA. We then employed the fluorescence resonance energy transfer (FRET) method to measure the effect of N-glycosylation site removal on the distance in the cell membrane between the EBOV-GP and HLA-I (HLA.A * 0201 allele). We recorded high FRET values for the interaction of CFP-fused HLA.A * 0201 and YFP-fused EBOV-GP, demonstrating the very close distance (<10 nm) between these two proteins on the cell membrane of GP-expressing cells. The co-localization of HLA-I and Ebola GP was unaffected by the disruption of steric shielding, as the removal of N-glycosylation sites on Ebola GP revealed similar FRET values with HLA-I. However, these mutations directed to N-glycosylation sites had restored immune cell function otherwise impaired due to steric shielding over immune cell ligands by WT Ebola GP. Overall, we showed that the GP-mediated steric shielding aimed to impair immune function is facilitated by the N-glycans protruding from its MLD and GCD domains, but these N-glycans are not controlling the close distance between GP and its shielded proteins.

show abstract

Mechanisms of Immune Evasion by Ebola Virus

Bhattacharyya

2021

Microbial Pathogenesis

View full text Add to dashboard Cite

The Ebola-Glycoprotein Modulates the Function of Natural Killer Cells

Cited by 22 publications

References 71 publications

Activating NK- receptors, homing selectins and inhibitory Siglecs recognize EBOLA-GP and HPV-L1NK

Activating NK- receptors, homing selectins and inhibitory Siglecs recognize EBOLA-GP and HPV-L1NK

N-Glycans Mediate the Ebola Virus-GP1 Shielding of Ligands to Immune Receptors and Immune Evasion

Mechanisms of Immune Evasion by Ebola Virus

Contact Info

Product

Resources

About