Journal: iScience

Article Title: Soluble CD4 inhibits Ebola virus infection by targeting endosomal receptor-binding site

doi: 10.1016/j.isci.2025.112573

Figure Lengend Snippet: Inhibition assay of sCD4 and CD4-derived protein molecules against pseudotyped and infectious Ebola viruses (A) sCD4 (2D), soluble first two-domain CD4; sCD4 (4D), soluble four-domain CD4; CD4-Ig, first two-domain CD4 linked with human Fc region (Ig domain) ; CD4-IgG2, first two-domain CD4 with human Fc region (Ig domain); EBOV, Zaire Ebola virus; BDBV, Bundibugyo ebolavirus. All experiments were conducted in triplicates and the error bars represent standard deviations from three experiments. (B) sCD4 (4D) inhibits infectious Ebola virus infection. This experiment was conducted in BSL-4 containment in triplicates, and the error bars represent standard deviations.

Article Snippet: Human soluble CD4 (4D) protein (CD4-3167H) was also purchased from Creative Biomart Inc. CD4-Ig Recombinant protein was obtained from NIH HIV Reagent Program.

Techniques: Inhibition, Derivative Assay, Virus, Infection

Journal: iScience

Article Title: Soluble CD4 inhibits Ebola virus infection by targeting endosomal receptor-binding site

doi: 10.1016/j.isci.2025.112573

Figure Lengend Snippet: Inhibition assay of CD4-mimetic small molecules against pseudotyped Ebola viruses (A) NBD-556 and analogs inhibiting EBOV infection. (B) Newly designed and synthesized compounds with phenyl ring substituted molecules in the region-I inhibiting EBOV. All experiments were conducted in triplicates and the error bars represent standard deviations from three experiments.

Article Snippet: Human soluble CD4 (4D) protein (CD4-3167H) was also purchased from Creative Biomart Inc. CD4-Ig Recombinant protein was obtained from NIH HIV Reagent Program.

Techniques: Inhibition, Infection, Synthesized

Journal: iScience

Article Title: Soluble CD4 inhibits Ebola virus infection by targeting endosomal receptor-binding site

doi: 10.1016/j.isci.2025.112573

Figure Lengend Snippet: Specificity assay of CD4-IgG-2 and JRC-II-191 against pseudo-typed HIV, EBOV, VSV, and A-MLV CD4-mimetic molecules CD4-IgG2 and JRC-II-191 were evaluated against four different viruses: HIV (YU2), EBOV, VSV (vesicular stomatitis virus), and A-MLV (amphitropic murine leukemia virus). All experiments were conducted in triplicates and the error bars represent standard deviations from three experiments.

Article Snippet: Human soluble CD4 (4D) protein (CD4-3167H) was also purchased from Creative Biomart Inc. CD4-Ig Recombinant protein was obtained from NIH HIV Reagent Program.

Techniques: Virus

Journal: iScience

Article Title: Soluble CD4 inhibits Ebola virus infection by targeting endosomal receptor-binding site

doi: 10.1016/j.isci.2025.112573

Figure Lengend Snippet: Binding affinity and kinetics assay of sCD4 (2D) and NBD-556 binding to EBOV receptor binding domain by biolayer interferometry Two-domain CD4 (sCD4-2D) (A) and NBD-556 (B) at two pH conditions, pH7.4 and pH6.1. Open circle in (B) was not included in the fit of the data. (See the BLI section for details).

Article Snippet: Human soluble CD4 (4D) protein (CD4-3167H) was also purchased from Creative Biomart Inc. CD4-Ig Recombinant protein was obtained from NIH HIV Reagent Program.

Techniques: Binding Assay

Journal: iScience

Article Title: Soluble CD4 inhibits Ebola virus infection by targeting endosomal receptor-binding site

doi: 10.1016/j.isci.2025.112573

Figure Lengend Snippet: Binding competition assay of sCD4 and CD4-mimetic compounds with receptor NPC1 (A) sCD4, NBD-556, JRC-II-191, and DY-III-228 competing with NPC1 receptor. (B) Dose response (0, 10, 20, and 40 μM) of NBD-556 in binding competition with NPC1 receptor. (C) Comparison of wild-type (EGPDCM-WT) and mutant (EGPDCM-mut) (WF/AA, 86W/A, and 88F/A) receptor binding domain (RBD) of EBOV binding to the NPC1 receptor. The mutant (WF/AA) surface expression level was confirmed to be comparable to the wild type (see ). Representative of 3 experiments.

Article Snippet: Human soluble CD4 (4D) protein (CD4-3167H) was also purchased from Creative Biomart Inc. CD4-Ig Recombinant protein was obtained from NIH HIV Reagent Program.

Techniques: Binding Assay, Competitive Binding Assay, Comparison, Mutagenesis, Expressing

Journal: iScience

Article Title: Soluble CD4 inhibits Ebola virus infection by targeting endosomal receptor-binding site

doi: 10.1016/j.isci.2025.112573

Figure Lengend Snippet: Molecular docking analysis of sCD4 and CD4mcs (A) Molecular docking analysis of sCD4 and CD4mcs binding to the EBOV-GP. sCD4 docking using HDOCK program. The docking energy is −150.41 kcal/mol. CD4mcs docking using AutoDock program. (a) sCD4-RBD ribbon model; (b) sCD4-RBD surface binding model; (c) sCD4-RBD interactions: hydrogen bond: GP T83:OG1-CD4 L44:N (green) and salt bridge: GP K84:NZ-CD4 D56:OD2 (brown) ; (d) NBD-556 docking ribbon model (−6.477 kcal/mol); (e) NBD-556 docking surface model; and (f), superimposed of NBD-556 (yellow), JRC-II-191 (magenta), and DY-III-228 (cyan). (B) Comparisons of two receptor binding sites of CD4bs and NPC1-bs. (a) CD4bs , of HIV gp120 (based on PDB 1G9N , YU2 strain). (b) NPC1bs , of EBOV-GP (based on PDB 5F1B , Zaire EBOV). Showing the accommodation of CD4-mimetic compound NBD-556 in the red dash circle. Hydrophobic surface in gray; CD4bs of HIV gp120 in magenta dash circle; NPC1bs of EBOV-GP in red dash circle.

Article Snippet: Human soluble CD4 (4D) protein (CD4-3167H) was also purchased from Creative Biomart Inc. CD4-Ig Recombinant protein was obtained from NIH HIV Reagent Program.

Techniques: Binding Assay