soluble human cd4  (Charles River Laboratories)

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

Journal: Neuroimmune Pharmacology and Therapeutics

Article Title: The link between chronic cocaine use, B cell perturbations, and blunted immune recovery in HIV-infected individuals on suppressive ART

doi: 10.1515/nipt-2022-0019

Figure Lengend Snippet: Increased plasma levels of anti-CD4 IgG in HIV+ cocaine users and its effect on CD4+ T cell death through ADCC. This study included HIV-negative cocaine users (HIV–/D) and non-users (HIV–/ND), and aviremic ART-treated HIV+ cocaine users (HIV+/D) and non-users (HIV+/ND). The median plasma anti-CD4 IgG levels (A) and CD4+ T cell counts (B) and their correlations in HIV+ subjects (C) were evaluated. (D) The ADCC was performed in CD4+ T and NK cells in the presence of 5 µg/mL of purified anti-CD4 IgGs from ART-treated HIV+ cocaine users (HIV+/D), anti-CD4 IgGs from ART-naive HIV+ cocaine users (Neg1), total IgGs (Neg2) from ART-treated HIV+ cocaine users and an anti-CD4 mAb (Pos). The percent of CD4+ T cytolysis was assessed using flow cytometry. Non-parametric Mann–Whitney U and Spearman correlation tests. *p<0.05; **p<0.01.

Article Snippet: Briefly, we coated the ELISA plate with human soluble CD4 protein (sCD4, Progenics, Tarrytown, NY).

Techniques: Purification, Flow Cytometry, MANN-WHITNEY

Journal: Infectious Agents and Cancer

Article Title: Antibodies to the CD4-binding site of HIV-1 gp120 suppress gp120-specific CD4 T cell response while enhancing antibody response

doi: 10.1186/1750-9378-3-11

Figure Lengend Snippet: Ab levels to whole gp120 and to CD4bs in sera of mice immunized with envelope proteins that have or lack CD4 binding activity a

Article Snippet: Soluble human CD4 (Progenics) was then added, and bound CD4 was detected using a mouse anti-human CD4 MAb OKT4 and goat anti-mouse IgG Abs conjugated to alkaline phosphatase.

Techniques: Binding Assay, Activity Assay

Journal: Infectious Agents and Cancer

Article Title: Antibodies to the CD4-binding site of HIV-1 gp120 suppress gp120-specific CD4 T cell response while enhancing antibody response

doi: 10.1186/1750-9378-3-11

Figure Lengend Snippet: Proliferative response of a murine CD4 T cell clone to gp120 is suppressed when gp120 is bound by anti-CD4bs mAbs . A) Mouse T cell clone T1 was tested for proliferation to different concentrations of gp120, alone or in the presence of anti-CD4bs mAb 654 or anti-C5 mAb 450 (10 μg/ml each). B) Proliferation of clone T1 was tested in response to different gp120/mAb complexes. For comparison, the response to gp120 alone is normalized to 100%. Gp120 was tested at 3 μg/ml and combined with 10 μg/ml of each mAb. Irradiated splenocytes were used as antigen-presenting cells. Average values and standard deviation from two repeated experiments are shown.

Article Snippet: Soluble human CD4 (Progenics) was then added, and bound CD4 was detected using a mouse anti-human CD4 MAb OKT4 and goat anti-mouse IgG Abs conjugated to alkaline phosphatase.

Techniques: Irradiation, Standard Deviation